Int. J. Environ. Sci. Tech., 4 (2): 223-232, 2007 ISSN: 1735-1472 © Spring 2007, IRSEN, CEERS, IAU
G. Ajeagah, et al.
Enumeration of Cryptosporidium sp. and Giardia sp. (oo)cysts in a tropical eutrophic lake 1*
1 2
G. Ajeagah; 1T. Njine; 1S. Foto; 1C. F. Bilong Bilong; 2P. Karanis
General Biology Laboratory, Faculty of Science, P. O. Box 812, University of Yaounde 1, Cameroon
National Research Center for Protozoan Diseases, Obihiro University for Agriculture and Veterinary Medicine, Japan Received 26 December 2006; revised 30 February 2007; accepted 10 March 2007; available online 20 March 2007
ABSTRACT: The biodynamic of Cryptosporidium sp. oocysts and Giadia sp. cysts have been assessed in the municipal lake of Yaounde, which is an artificial eutrophic lake that has been constructed on the Mingoa stream of the Mfoundi River Basin of Cameroon. The oocysts were identified by the Ziehl-Neelsen method while the cysts were identified by the Lugol iodine coloration. The values obtained suggest an increase in (oo)cysts density from the surface to the bottom of the lentic ecosystem. The oocysts varied from 56 oocysts/L in the upper layer to 2640 oocysts/L of water in the lowest layer, while the cysts population dynamics ranged from 24 cysts/L at the surface layer of the lake to 1713 cysts/L at the lowest layers of the lake. There was a considerable reduction in the resistant forms of these emerging pathogenic protozoa between the surface water of the Mingoa stream entering the Lake and that coming out from the Lake. The highest value of Cyptosporidium oocysts determined at the entrance of the lake is 1480 oocysts/L while at the outlet the highest value of oocysts identified is 620 oocysts/L. The highest value of Giardia cysts determined at the entrance of the lake is 352 cysts/L while at the outlet the highest value recorded is 294 cysts/L. Key words: Cryptosporidium, oocysts, giardia, cysts, municipal lake
INTRODUCTION Cryptosporidium sp. oocysts and Giardia sp. cysts have been reported to be ubiquitous in surface water (Leclerc, et al., 2002). Field surveys have also revealed the presence of low levels of oocysts and cysts in underground water (Watanabe, et al., 2005). This brings into the light the importance of microbiological and parasitological criteria for the quality control of recycled water that is being made available for potable water production and recreational usage (Katz, et al., 2006). In highly industrialized countries the most common human pathogenic protozoa transmitted by water belong to the genera Giardia and Cryptosporidium (Nygard, et al., 2006). Parasitic forms are transmitted by the faecal-oral pathway, with food and water being the passive career of the cysts and oocysts (Spinelli, et al., 2006). Giardiasis and Cryptosporidiosis are also common infections of domestic and wild animals which shed a relatively large number of cysts and oocysts in the environment (Gomez-Couzo, et al. 2005). These resistant forms of the protozoa are insensitive to disinfectants at the concentration commonly *Corresponding author, Email:
[email protected] Tel./Fax: +237 591 657
discharged in water treatment plants to reduce the pathogenic contamination of drinking water systems, even though it has been observed that at a higher concentration of chlorine and ozone, the cysts of Giardia sp. are less resistant than the oocysts of Cryptosporidium sp. (Finch, et al., 2002). Giardia cysts have been shown to survive in water for up to 2 months at temperatures as low as 8 °C, while Cryptosporidium oocysts can survive for up to one year at 4 °C in artificial sea water (Graczyk, et al. 2006; Nasser, et al. 2006). A large number of waterborne outbreaks with both protozoan have been reported worldwide (Karanis, et al., 2006). Giardiasis is a gastro-intestinal illness that is being caused by the flagellated protozoan Giardia intestinalis (G. lamblia or G. duodenalis). Some of the symptoms last for a long time. The infection rate for giardiasis is predicted as highest in developing countries (Ford 2006). This disease is highly correlated to the Human Immune Virus (HIV), which is at the origin of the immune deficiency of the human hosts, and thus the cause of an increase in the prevalence of this parasite (Guk, et al., 2005).
G., Ajeagah, et al.
The clinical symptoms of cryptosporidiosis are a watery diarrhea, abdominal pain, cramps, nausea and weight loss (Gatei, et al., 2006). This disease has gained a high prevalence in recent times because of the immunodepression of their host as a result of the HIV infection (Jong-Kyu, et al., 2005). It is a remarkable zoonotic pathogen, because it infects man a nd ot h er ver tebra t es (Gia n ga sper o, 2006). Cryptosporidium has been reported in rural and urban populations of the six continents. There is an increase risk of infection with this protozoon for persons who are exposed to surface water (Coupe, et al., 2006). There have been a lot of studies on the biodynamic of the resistant forms of these emerging pathogens in water (Ajeagah, et al., 2005, Smith and Thompson, 2001). Most of these analyses have been carried out on st r eam s. Th e qua n t it a t i ve da ta on t h e i den t i fica t i on of oocysts a n d cyst s i n la ke ecosystems and their survival in a lentic environment is still unavailable in developing countries in general and Cameroon in particular. This hydrosystem could be considered as a significant reservoir of (oo)cysts, that may be resuspended in the water column and
give rise to sporadic increases in the oocysts and cysts concentration in the whole river basin in which is situated the lake, resulting in the outbreaks of giardiasis and cryptosporidiosis. The aim of the present research was to evaluate the Giardia spp. cysts and Cryptosporidium spp. oocysts in a tropical ecosystem, which is the municipal lake of Yaounde (Cameroon). MATERIALS AND METHODS This study has been carried out in the Yaounde municipal lake which is a eutrophic aquatic ecosystem that is positioned in a NW-SE direction. It results from a dam that has been constructed on one of the tributaries of the Mfoundi River Basin, known as the Mingoa stream. The total surface area of the lake is 7.95X104 m2; it has an estimated volume of 19.104 m3 and a maximum depth of 4.3 m, while the mean depth is 2.4 m as presented on the bathymetry scale. (Fig. 1). The climate of Yaounde is equatorial, hot and humid, it is characterised by moderate precipitations (annual pluviometry mean of 1576 mm) and temperature has a low mean variation with time
Ekozoa
Verte
Abiergue Messa Lake Mfoundi 2
University
Chu
Mingoa
Olezoa Osogui Mfoundi 3
Akeu
Biyeme Mfoundi Mainstream 0
250
500
km
Fig. 1: Map of the Yaounde hydrosystem, indicating the location of the municipal lake 224
G., Ajeagah, et 4al.(2): 223-232, 2007 Int. J. Environ. Sci. Tech.,
(Suchel, 1987). The highest daily mensual thermic amplitude which is 10.4°C is recorded in February, while the lowest value which is 7.2°C is recorded in the month of July. December, January, July and August are the sub arid months, while April, May, September and October are the months with highest rainfall. There exist four seasons which are a long dry season from mid November to mid March, a short rainy season from Mid March to June, a short dry season from July to mid August and a long rainy season from mid August to mid November. The following study was carried from February 2002 to November 2003.
Five sampling points were chosen, which are two points on the Mingoa stream at the Inlet and the Outlet. Three points were selected inside the lake at 0 m of depth, 1 m of depth and 2 m of depth. Ten liters of water were collected from each of the points, at the 0 m, 1 m depth and 2 m depth. A Van dorn bottle of 6 liters of volume was used in the sampling of water for subsequent analysis. The samples were transported to the General Biology Laboratory of the University of Yaounde, where biological and physico-chemical analysis were carried out for the isolation and identifification of Cryptosporidium
Cryptosporidium oocysts/L
3000 2500 2000 1500 1000 500 0 F e brua ry
April
J une
Augus t
Oc to be r
Sample periods (2003) Lake-1
Lake-2
Lake-3
Fig. 2: Abundance dynamics of Cryptosporidium spp. oocysts identified in the various layers of the municipal lake 1600 Cryptosporidium oocysts/L
1400 1200 1000 800 600 400 200 0 F e brua ry
M a rc h
April
May
J une
J uly
Augus t
S e pte m be r
Oc to be r
No ve m be r
Sample periods (2003) Mingoa-1
Mingoa-2
Fig. 3: Abundance dynamics of Cryptosporidium spp. oocysts identified in the Mingoa stream at the Inlet and the Outlet of the municipal lake of Yaounde 225
G., Ajeagah, et al. Enumeration of Cryptosporidium spp and Giardia spp (oo)cysts in a tropical eutrophic lake...
spp., the water was allowed to settle for 24 hours in the laboratory. Zi nc sul ph a t e sol uti on was a dded t o t h e concentrated sample and then centrifuged.The supernatant was then concentrated in tubes in the presence of distilled water after a process of centrifugation. The slides were prepared with the deposit. The oocysts were then fixed with methanol and then coloured with basic fuchsine. The slides were rinced with dilute sulphuric acid and counter colour ed wi t h m a l a ch i t e gr een sol ut ion . Cryptosporidium spp. oocysts were then observed and enumerated under the Olympus light microscope at the 400x and 1000x magnification. Giardia cysts
were detected from the water by a concentration of particles contained in a 10 L sample. The samples were collected weekly from the sample points. The samples were allowed to settle in the laboratory. The deposit was distributed into centrifuge tubes and then fixed with formaldehyde. The samples were then centrifuged and zinc sulphate was added to the tubes to ensure a re-suspension of the Giardia cysts for eventual coloration with a solution of Lugol iodine. The cysts were observed under the Olympus microscope at the 400x and 1000x magnification. The ammonium nitrogen was analysed by the Nessler method, turbidity and suspended solids were assessed by a direct reading of the water sample on the spectrophotometer.
1800 1600
Giardia cysts/L
1400 1200 1000 800 600 400 200 0 F e brua ry
April
J une
Augus t
Oc to be r
Sample periods (2003) Lake-1
Lake-2
Lake-3
Fig. 4: Concentrations of Giardia spp. cysts/L identified in the various layers of the municipal lake of Yaounde
Giardia cysts/L
500 450 400 350 300 250 200 150 100 50 0 F e brua ry
April
J une
Augus t
Oc to be r
Sample periods(2003) Mingoa-1
Mingoa-2
Fig. 5: Concentrations of Giardia spp. identified in the Mingoa stream, at the inlet and outlet of the municipal lake of Yaounde, Cameroon 226
G., Ajeagah, Int. J. Environ. Sci. Tech.,et4al. (2): 223-232, 2007
Fig. 7: Microphotography of Cryptosporidium spp. oocyst
Fig. 6: Microphotography of Giardia spp. cyst
Month/Points February March April May June July August September October November
Table 1: Values of suspended solids (mg/L) in the Mingoa stream and the municipal lake Mingoa 1 (inlet) Lake 1(0 m) Lake 2 (1 m) Lake 3 (2 m) Mingoa 2 (Outlet) 125 40 60 85 40 1500 50 54 237.5 125 100 37.5 100 137.5 87.5 75 262.5 325 337.5 312.5 180 137.5 325 350 360 137.5 319 348 387 222 121 317 320 337 211 125 115 150 200 50 300 113 120 172 68 325 119 125 198 114
Table 2: Values of turbidity (FTU) in the Mingoa stream and the municipal lake Month/Points February March April May June July August September October November
Mingoa 1 (inlet) 1550 150 120 87.5 189 125 110 150 430 435
Lake 1 (0 m) 55 50 50 50 130 87.5 118 120 113 119
Lake 2 (1 m) 70 175 100 106.5 150 150 120 130 117 140
RESULTS Cryptosporidium spp. oocysts and Giardia spp. cysts have been isolated and identified in the municipal lake of Yaounde. The oocysts are round in structure (Fig. 7) with a double wall with dimensions ranging from 4-6 micrometers while the cysts (Fig. 6) are oval in structure with the presence of an axostyle, and dimensions ranging from 7-12 micrometers. The results obtained indicate the values for Cryptosporidium oocysts, Giardia cysts and the physico-chemical variables assessed. There is a
Lake 3 (2 m) 120 275 150 125 175 637.5 185 190 179 198
Mingoa 2 (Outlet) 106 175 75 112.5 112.5 150 40 45 110 114
higher concentration of Cryptosporidium oocysts at the inlet of the lake, with respect to the outlet. The number of oocysts counted increase with the depth. The values of Cryptosporidium oocysts identified range from 205 oocysts/L to 1480 oocysts/ L in the month of August and February respectively. At the outlet of the lake (Mingoa 2), the values range from 40 to 495 oocysts/liter in the months of July and March respectively. Inside the lake, the highest value recorded is 2720 oocysts/L in the month of April at the 2 meters depth, 227
G., Ajeagah, et al.
ammonium ion with respect to the outlet points assessed (Table 3).
while the lowest value obtained is 88 oocysts /liter in the month of September at the surface of the lake (Figs. 2 and 3). The values of the Giardia cysts dynamics follow th e sa me t rend l ike th ose of t he oocysts of Cryptosporidium (Figs. 3 and 4).There is a relatively higher density of cysts at the inlet of the lake, with respect to the outlet. The values obtained reveal that the maximum number of cysts at the inlet was 352 cysts/L in the month of April, while the lowest was 33 cysts/liter in the month of September. In the Mingoa 2 point, the highest cysts recorded were 452 cysts/liter in the month of May, while the lowest was 10 cysts/liter in the month of October. There was a relative increase in cysts density from the surface to the bottom of the municipal lake. The highest values of cysts determined was 1713 cysts/ L at the 2 m level in the month of May, while the lowest value obtained was 24 cysts/L at the surface, during the month of September. The values of turbidity and suspended solids increase from the surface to the bottom of the lake. The highest values of turbidity are 637.5 FTU at the lowest point of the lake analysed, this is during the period of July while the lowest value was 50 FTU at the surface of the lake, during the period from March to April. There were more suspended solids at the inlet of the lake with respect to the outlet. The suspended matter varied from a maximum value of 1500 mg/L at the entrance of the lake to 50 mg/L at the outlet of the lake in the months of September (Tables 1 and 2). Ammonia values in the study points ranged from 0.25 mg/L to 4.0 mg/L. There was an increase in the tenor of the ammonium ions from the surface to the bottom layers of the municipal lake. The inlet was generally charged with a higher content in the
DISCUSSION AND CONCLUSION The (oo)cysts are transported into this eutrophic ecosystem by the Mingoa stream which crosses m a ny i n ha bi t ed qua rt er s of t h e Ya oun de neighbourhood. It also receives untreated waste from the Messa residential quarter non-functional water treatment plant. The prevalence of Cryptosporidium spp. oocysts and Giardia spp. cysts during the rainy season is an indication that contaminated water may be important in its distribution as reported by Robertson, et al. (2006). The overall water sanitation and hygiene related death (99.8 percent) occurs in developing countries and 90 percent are deaths of children (WHO, 2003). Giardiasis and cryptosporidiosis are waterborne diseases that are spread through water in which water acts as a passive carrier for the transmissive forms of the infecting pathogens. These diseases depend also on the sanitary and environmental conditions of the region in question (Ajeagah, et al. 2005; Cifluentes et al. 2006). The protozoan pathogen C. parvum has been considered as a leading cause of waterborne gastroenteritis (Santos, et al. 2004). Due to its small size (5 µm), low specific gravity (1.05 g/ c3) and negative surface charge, C. parvum oocysts a r e gen er a l l y con si dered t o m ove t h r ough watersheds from their areas of emission to drinking water reservoirs with little attenuation of their population density in the stream channel (Marly et al. 2001), however the transport of oocysts in a lentic medium may be mediated by interactions with suspended sediments and by subsurface filtration and removal in streambed sediments (Dia & Boll 2006). The application of settling columns and flame
Table 3: Values of ammonium nitrogen (mg/L) in the Mingoa stream and the municipal lake Month/Points February March April May June July August September October November
Mingoa 1 (inlet) 1.5 1.0 2.9 1.6 1.5 1.38 2.66 4.0 3.6 3.5
Lake 1 (0 m) 0.4 0.25 0.25 1.0 0.63 1.13 1.59 1.36 1.25 1.50
Lake 2 (1 m) 0.15 1.0 0.5 2.2 0.75 1.35 1.96 2.41 2.65 3.05
228
Lake 3 (2 m) 0.55 1.35 0.90 3.40 1.00 2.13 3.90 2.96 3.50 3.90
Mingoa 2 (Outlet) 0.35 1.0 0.25 2.25 0.61 1.01 1.51 1.27 1.35 1.40
G., Ajeagah, et 4al.(2): 223-232, 2007 Int. J. Environ. Sci. Tech.,
recorded for the suspended solids, the turbidity and the tenor in ammonia from the surface water of the lake to the bottom layers. This same trend that has been observed for the oocysts and the cysts suggest the possibility of an adsorption model between the (oo)cysts and the particles that are present in the water column, which necessitate the transportation of the biomolecule inside the aquatic system. There is therefore the presence of receptor sites on the oocysts and cysts which are responsible for the binding with the organic and inorganic matter present in water and their consequent displacement with the solvent like an entity. The lake subsequently acts as a reservoir of pathogenic organisms which are likely to accumulate at the bottom sediments (Kistemann et al., 2002). These micro-organisms are a risk to public health, taking into consideration the fact that the Mingoa stream is one of the tributaries that is exploited for potable water production in the city of Yaounde, the water is also being used for recreational activities and fishing which places the population at a high health risk. This can be at the origin of epidemic episodes (Giovanni et al., 2006). The dysfunction of the waste water treatment systems places the population at increasing clinical risks as the protozoa G. lamblia, the zoonotic C. parvum and the anthroponotic C. hominis are the major causes of enteric diseases throughout the world (Guy et al. ,2006, Diaz et al. 2003, Hunter & Thompson 2005). The absence of a garbage treatment mechanism has been known to expose many children and the immunodepressed to severe gastroenteritis in the community (Diaz et al., 2003). These parasites are hyper endemic in areas that are lacking waste water and a drinking water treatment plant (Redlinger et al., 2002). The intensity of environmental stressors is geographically specific and may have different effects on the survival and persistence of pathogens in the environment (Alarcon et al., 2005). The reclaimed waste water does pose a significant health risk to humans due to the high pathogens levels and the lack of an effective disinfectant methodology (Geurden et al., 2005). There is a similar variation trend between water turbidity and the oocysts biodynamic in the various layers of the lakes (Sarco et al., 2006), as well as the relationship with the suspended solids which are
experiments have demonstrated that C. parvum oocysts attachment to several inorganic and organic sediments under varying water quality conditions may drastically influence the effective in-stream settling velocity of oocysts in natural mediums (Hijnen et al. 2005). These hydrostatic interactions with debris that are present in water could vary the settling velocity of the protozoan in the lake ecosystem. The rate of the downward transport on the oocysts concentration and reduction within the water column is between one and three orders of magnitude less than that caused by advection and di lut i on, depen di ng on th e str en gth of t h e hydrodynamic forcing in the water column (Brookes et al. 2006). The (oo)cysts are transported and deposited in the water column by mechanisms of attachment, detachment and straining (Bradford, 2005). Their transfer from the surface to the water sediments influences their movement in the lake ecosystem. Hydrodynamic interactions between an overlying flow and a sediment bed cause the (oo)cysts to accumulate in the sediments and reduce their concentration in the uppermost layers of the municipal lakes. Their association with other suspended particles increases both their effective settling velocity and the rate at which they are transferred to the sedimentary layer (Karim et al. 2004). This pathogen –sediment interaction plays an important role in regulating the concentration of these (oo)cysts in the hydrosystem (Searcy et al. 2006). The organic particles in water play an important role in the transportation of these emerging pathogens according to Medema et al. (1998), these (oo)cysts are generally linked to suspended particles in the water column. This adsorption influences the speed of sedimentation of the protozoa in the aquatic medium; this also increases the difficulties of purification of these parasites by water treatment plants (Weiss et al., 2005). The rapid urbanization of the city of Yaounde, places on the population an inadequate water supply a n d i n suffi ci en t sa n i ta r y resources, t h i s i s associated with increased human activity and this renders the lake highly polluted and a resultant accumulation of pathogens (Ajeagah et al., 2006). This is manifested by an increase in the values
229
Enumeration of Cryptosporidium sppG.,and Ajeagah, Giardiaetspp al. (oo)cysts in a tropical eutrophic lake...
necessary for the removal of the oocysts (Hu et al., 2004). The Yaounde municipal lake acts like a purification station because it reduces the density of parasites between the inlet and the outlet of the Mingoa stream. This leads to an accumulation of these enteropathogens in the lower layers of the municipal lake which can easily be released into the whole of the Mfoundi river basin of Yaounde, which can be at the origin of water related epidemics, in the cases where there is a perturbation of the water system, either by heavy rainfall or by other human activities. The high values of suspended solids and the consequent increase in the levels of turbidity in the municipal lake as presented in tables 1 and 2 reveals the possibility of anthropogenic effects on this eutrophic ecosystem. These actions can be at the origin of water related outbreaks of diarrhoea, for the entire community which relies on this water system for potable water production (Standish-Lee & Loboschefsky, 2006). High concentrations of the ammonium ions in the medium can probably increase the inactivation of the oocysts by their ability to penetrate into the walls of the oocysts and the membranes of the sporozoites. The maximum tenor in the ammonium ion in the municipal lake is 3.90 mg/L. The penetration of this ion into the double layers of the oocysts is facilitated by the pH values in the medium which are not adapted to the viability of the emerging parasites (Jenkins et al., 1998). The presence of some chemical elements such as sodium, chlorine and potassium ions reduces the process of excystation of the (oo)cysts (Kato et al., 2001). The Apicomplexan and the Sarcomastigophora isolated and identified in the water system are capable of contaminating the public drinking water supply, as well as the water used for recreational purposes (Collick et al., 2006). Cryptosporidium spp. oocysts and Giardia spp. cysts have been isolated and identified in the municipal lake of Yaounde. The results reveal an increase in (oo)cysts density from the surface layers to th e bottom dept h of th e l ake. T here is a considerable decrease in the biodynamic of the resistant forms of these pathogens between the inlet and the outlet of the lake. This suggests the characteristic purification properties of this lentic ecosystems and the application of this mechanism in sedimentary basins of water treatment plants. The
lake acts as a reservoir for Cryptosporidium oocysts and Giardia cysts in the ecosystem. The same trend recorded between the turbidity, the suspended solids with the (oo)cysts population dynamics suggests the possibility of an adsorption model in the transportation of the transmissive forms of the parasites in the lake ecosystem analysed. The accumulation of pathogenic micro-organisms at the bottom layer of the municipal lake exposes the population to an outbreak of giardiasis and/or cryptosporidiosis in cases that the resistant forms of the parasites are released into the entire river basin by fl oods i n t h e even t of hea vy r a i n s or anthropogenic activities. ACKNOWLEDGEMENT This study has been carried out with the material assistance of AIRE Development. Grants from this project have been used in the purchase of reagents and equipment for the detection of the pathogens and the physico-chemical analysis of the samples. REFERENCES Ajeagah, A.G., Foto, S. M., Njine, T., Wouafo, M., Moyou, R.S., Nola, M., Monkiedje, A., Ngassam, P., Zebaze, S.T., Kemka, N., (2005). Distribution of the Giardia sp cysts in the Mfoundi Wa ter ba sin (Ca meroon): influ ence of some physico-chemical factors of the mediu m. J. Ca m. Acad. Sci., 5, 85-90. Ajea ga h, G.A., Njine, T., Foto, S., Nola , M., (2 0 0 6 ). Assessment of Cryptosporidium spp oocysts in some urban wa ter systems in Ya ou nde (Ca meroon). Ca mer. J. Experiment. Biol., 2 , 9-1 5. Alarcon, M.A., Beltran, M., Ca rdenas, M.L., Ca mpos, M. C., (20 05 ). Presence and viability of Giardia spp. and Cryptosporidium spp in drinking water and waste water in the high basin of Bogota river. Biomed., 25, 353-65. Arnone, R.D., Wa lling, J.P., (2 0 0 6 ). Eva lu a ting of Cryptosporidium and Giardia concentrations in combined sewer overflow. J. Water Health, 4, 157-65. Bradford, S.A., Bettahar, M., (2005). Straining, attachment and detachment of Cryptosporidium oocysts in saturated porous media. J. Environ. Qual., 3, 469-78. Brookes, J.D., Da vis, C.M., Hipsey, M.R., Antenucci, P., (2 0 0 6 ). Associa tion of Cry p to sp o rid iu m with bovine faecal pa rticles a nd implica tions for risk reduction by settling within water supply reservoirs. J. Wat. Health, 4, 8 7 -9 8 . Ciflu entes, E., Ata mo, U., Kenda ll, T., Brink a rd, J. & Scrimsha v, S., (20 06 ). Rapid a ssessment procedures in environmental sanitation research: a case study from the northern border of Mexico. Ca n. J. Pu blic Hea lth, 9 7 , 2 4 -8 . Collick, A.S., Fogarty, E.A., Ziegler, P.E., Wa lter, M.T., Bowma n, D.D., Steenhu is, T.S., (2 0 0 6 ). Su rviva l of Cryptosporidium oocysts in calf Housing Facilities in the
230
Int. J. Environ. Sci. Tech., 4 (2): 223-232, 2007 G., Ajeagah, et al. determination of protozoa,Cryptosporidium and Giardia in water using method 1623. J. Microbiol. Methods 58(3), 3 2 1 -3 2 5 . Hunter, P.R., T hompson, R.C.A., (2 00 5 ). The zoonotic tra nsmission of Gia rd ia and Cry p to spo rid ium. Int. J. Parasitol., 35, 1181-1190. Jong-Kyu, Lee, Hyeon-Je, Song & Ja e-Ran, Yu . ,(2 0 05 ) Prevalence of diarrhoea caused by Cryptosporidium parvum in non HIV patients in Joellana m-do, Korea . Korean J. Parasitol., 43, 111 -114. Karanis, P., Kourenti, C., Smith, H., (20 06 ). Waterborne tra nsmission of protozoan parasites: a review of worldwide outbreaks and lessons we learnt. J. Wat. Health, in press. Karim, M.R., Manshad, F.D., Karpiscak, M.M., Gerba, C.P., (2004). The persistence and removal of enteric pathogens in constructed wetlands.Wat. Res.38, 391-399. Katz, D.E., Heisy-Grove, D., Beach, M., Dicker, R., Matyas, B., (2 00 6 ). Prolonged outbreak of giardia sis with two models of transmission. Epidemiol. Infect., 29, 1-7. Kistema nn, T., Cla ssen, T., Kock , C., Da ngendorf, F., Fischeder, R., Gebel, J., Vacata, V., Exner M., (20 02 ). Microbial load of drinking water reservoir tributaries during extreme ra infa ll a nd run-off. Apl. Environ. Microbiol., 65, 2188 -2197. Leclerc, H., Schwartzbrod, L., Die-Cas, E., (2002) Microbial a gents associated with wa terborne disea ses. Crit. Rev. Microbiol., 28, 371-409. Marly, X., Chevalier, S., Bues, M., Schwartsbrod, J., Setevenon, O., (20 01 ). Experimental investigations and nu merica l modelling of Cryptosporidium parvum transport behaviour in aquifers.Wat. Sci. Technol., 43,109-16. Medema, G.J., Schets, F.M., Teunis, P.F., Ha velaar, A.H., (19 9 8). Sedimentation of free and a tta ched Cryptosporidium oocysts and Giardia cysts in water. Appl. Environ. Microbiol., 64, 4460-4466. Nasser, A.M., Tweto, E., Nitza n, Y., (2 0 06 ). Die off of Cryptosporidium parvum in soil and waste water effluents. J. Appl. Microbiol. Online Early. Nygard, K., Schimmer, B., Sobstad, Q., Walde, A., Tveit, I., Langeland, N., Hausken, T., Aavitstland, P., (2006). A large community ou tbreak of wa terborne Gia rdiasis-dela yed detection in a non endemic area. BMC Public Health, 6 , 1 4 1 -1 4 6 . Redlinger, T., Corella - Banid, V., Graham, J., Galindo, A., Avita, R., Ca rdena s, V., (2 0 02 ). Hyper endemic Cry p to spo rid ium and Gia rdia in households lack ing municipal sewer and water on the United States – Mexico border. Am. J. Trop. Med. Hyg., 66, 794-798. Robertson, L.J., Forberg, T., Harmansen, L., Gjerde, B.K., Alsvag, J.O., La ngeland, N., (20 06 ). Cryp to sp orid iu m parvum infections in Bergen, Norway during an extensive outbreak of waterborne Giardiasis in autumn and Winter 2004. Appl Environ. Microbiol., 72(3), 2218-2220. Santos, L.U., Bonatti, T.R., Cantusio, N.R., Franco, R.M., (2004). Occurrence of Giardia cysts and Cryptosporidium oocysts in activated sludge samples in Campinas, Brazil. Rev. Inst. Med. Trop. Sao Paolo., 46, 309-313. Sarco, C., Bianchi, M., Lorini, C., Burini, D., Berchielli, S., La nciotti E., (20 06). Remova l of Cryptosp oridiu m a nd Giardia in drinking water treatment in Tuscan area. Ann. Tg. 18, 117-126.
New York city Wa tersheds. J. Environ Qua l., 35 , 6 80 687. Coupe, S., Delabre, K., Pouillot, R., Houdrat, S., SanttillanaHa ya t, M., Derou in, F., (2 0 0 6 ). Detection of Cryptosporidium, Giardia and Entero cytozoon bieneusi in surface water, including recreational areas: a one year prospective study. FEMS Immunol. Med. Microbiol., 47, 3 5 1 -3 5 9 . Dai, X., Boll, J., (2006). Settling velocity of Cryptosporidium parvum and Giardia lamblia. Water Res., 40, 1321-1325. Diaz, E., Mondragon, J., Ramirez, E., Bernal, R., (20 03). Epidemiology a nd control of intestina l pa ra sites with nitrazoxanide in children in Mexico. Am. J. Trop. Med. Hyg., 68, 3 84-3 85. Finch, G.R., Belosevic, M., (2002). Controlling Giardia spp and Cryptosporidium spp in drinking water by microbial reduction processes. J. Envion. Eng. Sci., 1, 17-31. Ford, T., (2 0 0 6 ). Emerging issu es in wa ter a nd hea lth research. J. Water Health., 4, (suppl. 1), 59-65. Gatei, W., Wamae, C.N., Mbae, C., Waruru, A., Mulinge, E., Waithera, T., Gatika, S. M., Kamwati, S. K., Revathi, G., & Ha rt, C.A., (2 0 0 6 ). Cryptosporidiosis: Preva lence, genotype analysis and symptoms associated with infections in children in Kenya. Am. J. Trop. Med. Hyg., 75, 78-82. Geu rden, T., Cla errbou t, E., Vercru yesse, J., (2 0 0 5 ). Protozoan infection causes diarrhoea in calves. Tijdscchr Diergeneek d. 13 0, 7 34-7 37. Giangaspero, A., (2006). Giardia, Cryptosporidium and the spectre of zoonoses. The Italian experience from land to sea. Parassitologia, 48, 95-100. Giova nni, G.D., Beta ncou rt, W.Q., Herna ndez, N.W., Assa da ia n, J.P., Flores, M., Lopez, E.J., (2 0 0 6 ). Investiga tion of the potentia l of zooa nthroponotic transmission of Cryptosporidiosis and Giardiasis through a gricu ltura l u se of reclaimed waste wa ter. J. Environ. Health Res., 16, 6, 1-32. Gomez-Couzo, H., McLauchlin, J., Ares-Mazas, E., (2005) Characterisation of a Cryptosporidium isolate from water buffalo (Bubalus bubalis) by sequencing of a fragment of the Cry p to sp o rid iu m oocysts wa ll protein gene (COWP).Vet. Parasitol., 13 1, 139 -14 4. Graczyk, T.K., Girouard, A.S., Tamang, L., Nappier, S.P., Schwa h, K.J., (2 0 0 6 ). Recovery bioa ccu mu la tion a nd ina ctiva tion of hu ma n wa terborne pa thogens by the Chesapeake Bay non native oyster, Crassostrea ariakensis. Apl. Environ. Microbiol., 75, 3390-3395. Guk, S.M., Seo, M., Park, Y.K., Oh, M.D., Choe, K.W., Kim, J.L., Choi, M.H., Hong, S.T. Chai, J.Y., (2005). Parasitic infections in HIV infected pa tients who visited Seou l National University Hospital during the period 1995-2005. Korean J. Parasitol., 423, 1-5. Guy, A.R., Payment, P., Kru ll, U.J., Horgen, P.A., (200 6) Rea l time PCR for qua ntifica tion of Gia rd ia and Cry p to spo rid ium in environmental wa ter sa mples and sewage. Appl. Environ. Microbiol., 69, 5178-5185. Hijnen, W.A., Brou wer– Ha nzens, C.A.J., Medema , K.J., (20 0 5). T ransport of MS2 phage, Esc h erich ia c o li, Clo stridium perfringe ns, Cry ptosporid ium parv um a nd Giardia intestinalis in a gravel and a sandy soil. Environ. Sci. Technol., 39(20), 7860-7868. Hu, J.Y., Feng, Y.Y., Ong, S.L., Ng, J., Song, L.F., Tan, X.L., Chu , X.N., (2 00 4 ). Improvement of recoveries for the
231
G., Ajeagah, et al. Suchel, J.B., (1 987 ). Les climats du Camerou n. Thèse de Doctorat d’Eta t, Univ., Bordea ux 3 . France.118 6. Watanabe, Y., Kimura, K., Yang, C.H., Ooi, H.K., (2005). Detection of Cryp tospo rid iu m sp. oocysts a nd Giardia sp cysts in fa u cet wa ter sa mples from ca ttle a nd goa t farms in Taiwan. J. Vet. Med. Sci., 67, 1285-1287. Weiss, Z.J., Brouwer, E.J., Aboytes, R., LeChevalier, M.W., O’ Melia, C.R., Schwab, K.J., (2005). Riverbank filtration for control of micro-orga nisms, resu lts from field monitoring.Wa ter Research, 39 , 1 9 90 -2 0 01 . World Hea lth Orga nisa tion, (WHO), (20 0 3). Qu a ntifying selected major risks to health. The World Health Report 2002 , Geneva (Chapter 4).
Searcy, K.E., Packman, A.I., Atwill, E.R., Harter, T., (2006). Deposition of Cry pto sp oridium oocysts in strea mbeds. Apl. Environ. Microbiol., 72 , 181 0-181 6. Smith, M., Thompson, K.C., (2001). Cryptosporidium: The a na lytica l cha llenge. R. Soc. Chem. MPG Book s Ltd, Bodmin, Cornwall, UK. Spinelli, R., Brandonisio, O., Serio, G., Trerotoli, P.,Ghezzani, F., Carito, V., Dajci, N., Doci, A., Picaku, F., Dentico, P., (2006). Intestinal parasites in healthy subjects in Albania. Eur. J. Epidemiol., 6, 161-166. Standish-Lee, P., Loboschefsky, E., (2006). Protecting public health from the impa ct of body-conta ct recrea tion.Wa t. Sci. Technol., 53 , 2 01-2 07.
AUTHOR (S) BIOSKETCHES Ajeagah, G., M.Sc., DEA, Ph.D., Research student in hydrobiology and environment, University of Yaounde 1, Cameroon. Email:
[email protected] Njine, T., Doctorat de troisième cycle, Doctorat d’Etat, is a Professor of Zoology and Dean of the Faculty of Science in the University of Yaounde 1, Cameroon. Email:
[email protected] Foto, S., Doctorat de troisième cycle and Researcher for a Doctorat d’Etat, is a Senior lecturer in hydrobiology in the University of Yaounde 1, Cameroon . Email:
[email protected] Bilong Bilong, C.F., Doctorat de troisième cycle, Doctorat d’Etat, is a Professor of Parasitology in the University of Yaounde 1, Cameroon . Email:
[email protected] Karanis, P., Ph.D., is a researcher in the Unit for Molecular Epidemiology and Protozoan Pathogenetics, National Research Center for Protozoan Diseases, Obihiro University for Agriculture and Veterinary Medicine, Japan, Inada-cho, Obihiro 080-8555, Japan. Email:
[email protected]
This article should be referenced as follows: Ajeagah, G., Njine, T., Foto, S., Bilong Bilong, C.F., Karanis, P., (2007). Enumeration of Cryptosporidium spp and Giardia spp (oo)cysts in a tropical eutrophic lake: The municipal lake of Yaounde (Cameroon). Int. J. Environ. Sci. Tech., 4 (2), 223-232.
232
