Snails and fish as pollution biomarkers in Lake Manzala and laboratory a: Lake Manzala snails.
Lake Manzala is considered one of the most important lakes in Egypt. It is exposed to high levels of pollutants from industrial, domestic and agricultural resources [1-3]. Ali reported that Lake Manzala receives about 4000 million cubic meters of untreated industrial, domestic and agricultural waste water annually .
The use of physiological and biochemical parameters as indicators of water quality has been developed to detect sublethal impacts of pollutants. Prominent among these biomarkers are physiological variables, such as plasma levels of metabolites , haematological data [6,7], levels of hormones [8-11] and biochemical variables such as detoxifying enzyme activities [12,13]. Interesting reports concerning the mechanisms of metal uptake, accumulation, transport, and elimination of metals in molluscs are usually focused on chemical, biochemical, molecular, and physiological aspects [14-21]. El-Khayat assessed genetic variation and genetic pattern of Lymnaea snails collected from irrigation canals in four different Governorates using ISSR markers, with the characterization of environmental parameters of the collecting Lymnaea sites. The authors showed high polymorphism by using for the first time the ISSR PCR technique for studying genetic variations of L. natalensis snails in Egypt and concluded that L. natalensis snails can survive associated with other snails, plants, and insects and can be tolerate the heavy metals in water .
Similarly, histopathological changes have been widely used as biomarkers in the health evaluation of animal organisms. The discharge of toxic elements into the rivers, estuaries and coastal waters poses serious pollution and consequently affects the fish, flora and fauna as snail.
Moreover, freshwater molluscs play an important role in aquatic ecosystems, providing food for many fish species and vertebrates .
This work aims to record the alterations of the Physiological, hematological and histopathological parameters in snails collected from Lake Manzala as a bio-indicator for water pollution.
Materials and Methods
Snail samples were collected from 8 sites in Lake Manzalafrom 3 governorates; Port-Said (Kobry El-Lansh, Kaar El-Bahr and El-Khankak), Dakahlya (Gammalya, Matarya and Nasayma) and Damietta (Ananyya and Sayala). The snails collected were kept in water from their habitat and examined for natural infection. The negative (uninfected) Biomphalaria snails and other collected species (Physa and Planorbis) were contributed in the physiological studies. On the other hand, both negative and positive Biomphalaria samples were examined histologically.
Determination of liver and kidney functions: The assessment of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), urea, creatinine, total and direct bilirubin, albumin (ALB) and total protein were examined in snail tissue extracts. They were assayed biochemically using biosystem autoanalyzer, Backmann at Theodor Bilhaz Institute (TBRI) hospital laboratories. Snail tissues were dissected out, homogenized in bi-distilled water (1:1 w/v) using motor homogenizer and centrifuged at 5000 rpm for 20 min at 4[degrees]C and the supernatants were taken and kept at -20[degrees]C till used as described by .
Creatinine was determined according to . In this method, creatinine reacts with picrate to form a coloured complex and the rate of formation of the complex is measured photometrically at 492 nm.
Urea was determined by using the coupled urease/glutamate dehydrogenase (GLDH) enzyme system according to .
Determination of antioxidant enzymes: The antioxidant enzymes catalase (CAT), Glutathione-S-Transferase (GST) and Gamma Glutamyltransferase (GGT) were assayed in snail tissue extract using spectrophotometer. Snail's tissues were dissected out. Each snail tissue from each treatment was homogenized in bi-distilled water (10:1 w/v) using motor homogenizer. Homogenates were centrifuged at 5000 rpm for 20 min at 4[degrees]C and the supernatants were taken and kept at -20[degrees]C till used.
Determination of snail hemolymph components: Snail hemolymph was collected in accordance to the technique of . The hemolymph was obtained via small hole made in the shell into which capillary tube was inserted then it was drawn into tube by capillary suction. The hemocytes of the samples hemolymph were determined by haemocytometer. For total and differential counting, monolayer of hemocytes were stained with Giemsa stain for 20 minutes, according to the methods of  and counted by light microscopy.
Histopathological examinations: Snail specimens collected from Lake Manzala were dissected, removed from their shells gently and fixed in 10% buffered neutral formalin solution. Five-micron thick paraffin sections were prepared, stained by hematoxylin and eosin (HE) and then examined microscopically and photographed for histopathology observations .
Statistical analysis: Data are expressed as means [+ or -] SD. The results were computed statistically (SPSS software package, version 20) using the T-test analysis. Values of p<0.05 were considered statistically significant.
The present results showed significant increase in AST, ALT, and
ALP in Planorbis and Physa, respectively in most samples collected from Nasayma site in Lake Manzala. Most of snails collected from Port-Said and Dakahlya sites showed significant increase in urea. Results of creatinine in samples from different lake sites showed alteration, ranged between non-significant decrease and increase (Table 1). Significant increase of total protein level was obtained in all field samples while total bilirubin showed the highest levels in physa and planorbis samples collected from Nasayma, Dakahlya and in Biomphalaria samples collected from Matarya, Dakahlya. Also, results showed higher levels of indirect bilirubin than direct. Most snail samples showed approximately normal A/G ratio (Table 2).
Significant alterations in catalase (CAT) level were noticed in all snail samples collected from Lake Manzala as compared with lab bread controls (except in Planorbis collected from Kobry El-Lansh and Biomphalaria from Gammalya and Biomphalaria from Nasayma). The recorded alterations in the snail samples was increased by 18 to 185%, or decreased by -13 to -90% (Table 3).
Glutathione-S-transferase (GST) alteration was demonstrated in all samples includes decrease in activity ranging from -21% to -83% (P<0.001) and increase in activity ranging from 13% to 119%.
The same result was noticed in Gamma-glutamyl transpeptidase (GGT) in snail samples as compared with lab bread controls, some samples showed decrease change activity ranging from -1% to -35% and other samples showed increase change activity ranging from 6% to 666%, (Table 3).
Determination of hemolymph components
The majority of snail samples showed significant decrease in total and differential cell count as compared with lab bread controls (Table 4). The higher percent of decrease in the total cell count (-72%) was recorded in Biomphalaria collected from Nasayma, Dakahlya. Hemoglobin concentration showed alteration; increased to 2.6 g in Planorbis collected from Kobry El-Lansh and decreased to 0.8 g in Physa collected from Annanya, Damietta.
The histopathological observations
A knowledge of the normal histology and structure of snails is guided by .
Head foot: The normal foot region has an outer cuticular layer as a protective layer of the foot. Inner to this lining there is a tall columnar epithelium with basal nuclei in its cell. Amongst the columnar epithelium there are modified sacs like cells in the form of unicellular glands which open through the cuticular layer exterior to the foot surface. These unicellular glands are involved in mucous secretion. Embedded in between there are transversely muscle fibers, called as longitudinal muscle fibers. Major part of the foot muscles are made up of thickly arranged oblique muscle fibers.
Histopathological observations in foot region of Biomphalaria snail samples showed necrotic change (shrinkage) in the mucous secreting unicellular glands (Figure 1b) and hyaline substances are shown in samples collected from Port Said (Figure 1c) and splitting fiber tissues in Dakahlya and Damietta snails (Figure 1d,1e). Also, results showed oblique splitting muscle fibers, increased empty spaces and atrophy within muscles of snail head in Dakahlya samples (Figure 1f,1g,1h).
Salivary gland: The normal salivary gland of B. alexandrina snail composed of two lobs found in the buccal mass as shown in (Figure 2a).
The histopathological effects of polluted water showed shrinkage and atrophy in the salivary gland of snails collected from Damietta (Figure 2b), focal areas of necrosis (Figure 2c,2d), large fat vacuoles (Figure 2e) and enlargement of the salivary gland (Figure 2f) in snails collected from Port said.
Central ganglia: The central nervous system ganglia are in the form of compact mass of ring surrounding the esophagus of the snail. (Figure 3) showed that all ganglia exhibit presence of enlarged neurosecretory neurons (Figure 3a). Fibrosis (Figure 3b,3c) and degeneration with large vacuoles (Figure 3d) were observed in snail samples collected from Damietta and Dakahlya (Figure 3).
Hepatopancreas: The normal histological structure of Biomphalaria hepatopancreas includes glandular tubules interspersed with connective tissues. The entire gland is enclosed within a thin walled sac called as tunica propria. The hepatopancreatic epithelium is rested on thin basement membrane; at least 3-4 types of cells can be recognized in the hepatopancreatic epithelium of the snail, digestive, calcium and excretory cells (Figure 4a). The histopathological changes showed cellular necrosis followed by loss of secretory activity of the epithelial cells in Port Said samples (Figure 3b). Also, atrophy, degeneration and fat vaculation were noticed in Port Said and Damietta samples (Figure 4c,4d). Dilated lumen and more than two hepatopancreatic tubules connected together with one larger lumen in Dakahlya samples (Figure 4e,4f).
Male organs (Prostate gland): The normal histological structures of the male organs of B alexandrina composed mainly of sperm duct and the prostate tubules (Figure 5a).
The histopathological observations of Port Said samples showed severe dilated sperm duct and prostate tubules, dilated lumen of prostate tubules which filled with hyaline and degeneration wall with necrotic change (Figure 5b,5c). while Dakahlya samples showed enlarged sperm duct, degenerated prostate tubules and clogged sperms.
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The prostate gland in Damietta samples showed severe degeneration and atrophy (Figure 5b,5c).
The hermaphrodite gland: Histology of normal hermaphrodite gland of the adult B. alexandrina snails as that of any other pulmonate snail consists of number of vesicles known as acini separated from each other by thin vascular connective tissue (Figure 6a). Each acinus is enveloped in a sheath of squamous epithelium. In each acinus both male and female reproductive gametes are produced where mature ova are located at the periphery of the acini and bundles of male sperms are arranged in the center. Various stages of sperm and ovum development (simultaneous) are evident.
Histopathological alteration in Port Said samples included, acini lost their normal architechture and their separating connective tissues are almost degenerated (Figure 5b). The acinar epithilum showed necrotic changes in the form of decreasing cytoplasm of oocytes and partial destruction (Figure 6c). Atrophy and reduction in the number of sperms was also observed (Figure 6d). Degenerative changes were observed in most of the ova, where some of them have faint staining nuclei and others lost their nucleous (Figure 6e,6f). Some acini appear more or less evacuated and large fat vacuoles can be seen in Dakahlya samples (Figure 6g). Damietta showed the most degenerated features in the hermaphrodite gland as atretic oocytes and sperms and atrophy of most gland components (Figure 6h,6i).
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The infected Biomphalaria samples collected from lake manzala
Some of Biomphalaria samples collected from Dakahlya and Port Said showed the presence of parasite sporocycts.
The oblique muscle fiber got damaged and mother sporocysts take place within foot muscles, thereby causing splitting, necrosis and increased empty spaces within muscle fibers (Figure 7a-7c).
The digestive gland was destructed while daughter sporocycts which contain many developing cercariae were noticed. The histopathological changes of digestive gland of B. alexandrina induced exudation in the lumen of tubules, expansion of hemolymphatic spaces between the tubules, loosing of connective tissue and increase of vaculation and necrotic changes in the digestive cells (Figure 7d,7e,7f).
Accumulation of heavy metals in snail tissues
(Figure 8) showed the accumulation of heavy metals in head foot tissues of Biomphalaria snails collected from Port Said, Damietta and Dakahlya samples.
Under conditions of pollution mollusks are susceptible to the pathogenic effects of toxicants, which in turn may result in detrimental changes to their immunological and physiological processes .
The present results showed significant increase in AST and ALT, and ALP in Planorbis and Physa snail samples, collected from Nasayma site in Lake Manzala. Moreover, the results showed alterations in CAT, GST and GGT activity in snail samples collected from Lake Manzala.
AST and ALT are vital enzymes in the metabolism and generation of energy from amino acids . Therefore, the elevated transaminases may indicate the high energy demand of the snail under stressful conditions of intoxication. Also, the increase in ALT, AST and ALP enzymes were correlated with alteration in phospholipid metabolism  which indicated mainly to hepatocellular disorder . Under physiological stress conditions in animals, the catalytic activity of the urea pathway enzymes is also accelerated .
These results are in agreement with  who recorded a significant increase of transaminases activity and catalase in the garden snail specimens (Helix pomatia L) which were collected from polluted area compared to control. Also,  indicated that there are significant elevations in the levels of acid phosphatase and alkaline phosphatase, after using of Profenophos against B. alexandrina, which can be explained by the destruction of internal snail cells. Mohamed revealed an elevation in the activities of AST, AlT and AkP enzymes in snails' tissues post treatment with [LC.sub.10] and [LC.sub.25] of Basudin, Selecron and Bayluscide in comparison with control groups . Some other authors recorded increase of the activity of these enzymes, while others recorded decrease in intoxicated animals [36,39,40]. Abdel-Daim reported increased serum AST, ALT, ALP, cholesterol, urea, uric acid, creatinine and tissue MDA after application of deltamethrin subacute intoxication (1.46 [micro]g/L for 28 days) against Oreochromis niloticus fish . At the same time they found that tissue levels of GSH, GSH-Px, SOD and CAT were reduced. On the other hand,  recorded suppression of the antioxidant enzyme activity and alterations of serum biochemical parameters in freshwater fish Nile tilapia, Oreochromis niloticus.
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Significant increase of total protein level was recorded also in all samples collected from Lake Manzala. This increase may be attributed to the changes in hepatic protein synthesis [43,44] due to the stress in the polluted habitat. These results go in the same direction as those of  who recorded an increase in the total protein concentration in Helix snails dependent in the presence of metal dust. Also,  highlighted a significant increase in the total protein rate under the effect of a chemical stress at different biological models. Mello observed significant changes in protein metabolism in response to exposure to different concentrations of E. splendens var. hislopii latex, with significant increases in snails exposed to 0.8 and 1.0 mg/l of the latex, indicating latex toxicity . The same was observed by  using other plants and higher concentrations.
Snails collected from most Port-Said and Dakahlya sites showed significant increase in urea. Urea is only synthesized in liver from excess amino acids and excreted by kidney and major illness may increase urea levels . The variation in the nitrogen degradation products showed that the increase of urea content occurred when the uric acid level declined. In accordance with this, the exposure of Biomphalaria glabrata to Euphorbia splendens var. hislopii latex caused the urea content increased which reflects a disturbance in the snail's regulation of their metabolism due to intoxication caused by the latex exposure .
Snail samples collected from most sites of Lake Manzala showed significant decrease in total hemolymph cell count, hyalinocytes, round small hemocytes and granuolocytes. The decrease in hemolymph cells may be considered as a haemolysis response to the multiple pollution elements in Lake Manzala. This was mentioned by  that haemocytosis represents a response to external stress or certain stimuli and may originate from a variety of biotic or abiotic sources . These results were in agreement with  who found that exposure to dyestuff and chemical effluent could result in decreases in RBC count and Hb content which are symptoms of anemia.
The histopathological changes produced by pollutants in organs and tissues can occur before they produce irreversible effects on the biota. So, histological methods can be used in conjunction with other parameters and/or ecotoxicological bioindicators as an early warning system for the survival of the species, as well as for environmental protection.
Histopathological observations in head foot region of Biomphalaria snails showed shrinkage in the mucous secreting unicellular glands and hyaline substances in samples collected from Port Said sites, splitting fiber tissues, increased empty spaces and atrophy within muscles of snail head in Dakahlya and Damietta samples. The salivary gland of snails collected from Damietta showed shrinkage and atrophy while there were focal areas of necrosis, large fat vacuoles and enlargement of the salivary gland in snails collected from Port Said. All snails ganglia showed modified and enlargement of neurons, degeneration with large vacuoles and fibrosis in samples collected from Damietta and Dakahlya. The histopathological changes of hepatopancreas included cellular necrosis followed by loss of the epithelial cells were shown in Port Said samples. Also, atrophy, degeneration and fat vaculation were noticed in Port Said and Damietta samples. Dilated lumen and more than two hepatopancreatic tubules connected together with one larger lumen in Dakahlya samples. The prostate gland in snails of Port Said samples showed severe dilated sperm duct and prostate tubules, dilated lumen of prostate tubules which filled with hyaline and degeneration wall with necrotic change. While Dakahlya samples showed enlarged sperm duct, degenerated prostate tubules and clogged sperms. The prostate gland in Damietta samples showed severe degeneration and atrophy. Regarding the hermaphrodite gland in Dakahlya samples, decreasing cytoplasm of oocytes, partial destruction, lost nucleus, large fat vacuoles, atrophy and reduction in the number of sperms were observed. Damietta samples showed the most degeneration features in the hermaphrodite gland as atretic oocytes and sperms and atrophy of most gland components.
All these histopathological damages in snail organs may be due to the pollution of Lake Manzala water by heavy metals which recorded by . Stress responses in invertebrates can occur following acute or chronic exposures to contaminated environments and as such, the overall health status of individuals within those environments, both in terms of histopathological lesions and the presence of infecting organisms, may ultimately reflect the general health status of these sites .
The digestive glands of molluscs have been known as target organs for contaminant effects because; this organ plays a major role in contaminant uptake, intracellular food digestion and metabolism of inorganic and organic chemicals in the organisms [55-57]. However, particulate metal uptake is mainly achieved via the digestive tract by endocytosis; further metals are transferred first to lysosomes and then to residual bodies, especially in the digestive cells of the digestive gland . It could also be possible that in the damage in the snail's hepatopancreas including the alteration of liver and kidney enzymes is according to functionality analog with vertebrate's liver that accumulate mostly heavy metals compared to other organs, and which damage it also [59,60].
In agreement of these results, the exposure of the snails Archachatina marginata to sublethal concentrations of the metals resulted in a prevalence of hepatocellular foci of cellular alterations (FCA) in the hepatopancreas of snails. Basophilic adenoma and ovotesticular fibrillar inclusions were also observed in the ovotestes of snails exposed to the test metals . Jonnalagadda have been reported histopathological alterations such as degeneration and the gathering of amebocytes in areas between the tubules in the digestive gland of snail Bellamya dissimilis exposed to endosulfan . The histopathological examinations of Lymnaea luteola exposed to Paraquat (Gramoxone) revealed the following changes: amebocytes infiltrations, the lumen of digestive gland tubule was shrunken; degeneration of cells, secretory cells became irregular, necrosis of cells and atrophy in the connective tissue of digestive gland . Moreover, it is worthy to mention that in the freshwater snails nervous system has been proved to be sensitive to many toxic materials and cytotoxicants that may induce injurious consequences [64-66].
Some of Biomphalaria samples collected from Dakahlya and Port Said showed the presence of parasite sporocycts. The most histopathological deleterious effects have been noticed within the tissues caused in the foot and hepatopancreas due to the invasion of larval trematode parasites to the host snail B. alexandrina. The oblique muscle fiber got damaged may be due to penetration of miracidia at the time of infection in the nature. Since earlier stages of larval development i.e. sporocyst and mother sporocyst, takes place within foot muscles, thereby causing increased empty spaces within muscle fibers after their entry in to the viscera of the snail. The digestive tubules epithelium got damaged to the extent of loss of normal tubular structure may be due to metabolic and other excretory materials in the form of granules found scattered in the connective tissue. The destruction of the digestive gland was even more severe may be due to the developing of daughter sporocycts which contains many of the developing cercariae.
Similar observations were recorded by  in the snail Archachatina marginata that the digestive gland tubule becomes compressed thereby resulting reduced tubular lumen of the gland as observed by that more cercaria and rediae were found in between the hepatic tubules and tunica propria causing extension of the space between tubules.
The histological observations of Biomphalaria snails collected from Lake Mazala showed accumulation of heavy metals in the head foot tissues. This was proved in the study of  who recorded that the metals concentrations were higher in snail tissues and water samples from Lake Manzala. The collected water samples from Damietta sites showed the highest significant Cu & Cd concentration while Port-Said samples showed the highest Pb concentration and Dakahlia showed the highest Zn concentration.
In conclusion, the severe alterations and degeneration recorded in the physiological and hematological parameters and also histopathological observations are clear evidence for the pollution of the water from which these snail samples were collected. This conclusion is confirmed by  who recorded highly significant concentrations of Cu, Cd, Pb and Zn in water samples from different Lake Manzala sites. Also, these metals were highly concentrated in snail and fish tissues and the higher metal bioaccumulation was determined in snails collected from sites showed higher water metals concentrations.
Abbrevations: AST: Aspartate Aminotransferase; ALT: Alanine Aminotransferase; ALP: Alkaline Phosphatase; ALB: Albumin; CAT: Catalase; GST: Glutathione-S-Transferase; GGT: Gamma Glutamyltransferase; TBRI: Theodor Bilhaz Institute; HE: Hematoxylin and Eosin Stain; A/G ratio: Albumin/Glubuline ratio
This study is a joint project (Biomarkers as indicators of environmental pollution: Experimental approach and case studies), kindly funded by the Academy of Scientific Research and Technology through the Bilateral Agreement between Academy of Scientific Research and Technology of the Arab Republic of Egypt and Bulgarian Academy of Sciences (2012-2014).
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[67.] El-Khayat HMM, Mahmoud KMA, Gaber HS, Abdel-Hamid H, Abu Taleb HMA (2015b) Studies on the effect of pollution on Lake Manzala ecosystem in portsaid, damietta and Dakahlya governorates, Egypt. J. Egypt. Soc. Parasitol. (JESP). 45: 155-168.
Hanaa MM El-Khayat (1), Hoda Abdel-Hamid (1), Hanan S Gaber (2), Kadria MA Mahmoud (1) and Hassan E Flefel (1)
(1) Department of Environmental Researches and Medical Malacology, Theodor Bilharz Research Institute, Imbaba, PO Box-30, Giza, Egypt
(2) National Institute of Oceanography and Fisheries, Cairo, Egypt
* Corresponding author: Hanaa MM El-Khayat, Department of Environmental Researches and Medical Malacology, Theodor Bilharz Research Institute, Imbaba, PO Box-30, Giza, Egypt, Tel: 202 354 01019; E-mail: email@example.com
Received September 29, 2015; Accepted December 11, 2015; Published December 16, 2015
Table 1: Aspartate amino transferase (AST), alanine amino transferase (ALT) alkaline phosphatase (ALP), glucose, creatinine and urea in tissue extract of snails collected from Lake Manzala. Parameter AST (Unites/ml) Treatments Mean [+ or -] Change SD % Control lab 21.9 [+ or -] 4 Bort-Said Kaar Biomphalaria 26.2 [+ or -] 2 20 El-Bahr Planorbis 30.2 [+ or -] 4 38 El-Khankak Biomphalaria 11 [+ or -] 0 ** -50 Kobry Biomphalaria 37.4 [+ or -] 5 * 71 El-Lansh Planorbis 40.5 [+ or -] 6 * 85 Dakahlya Gammalya Biomphalaria 31.1 [+ or -] 0 ** 42 Nasayma Biomphalaria 29.6 [+ or -] 3 * 35 Planorbis 56.4 [+ or -] 2 ** 158 Physa 53.8 [+ or -] 3 ** 146 Matarya Biomphalaria 32.1 [+ or -] 2 * 47 Damietta Annanya Physa 34.2 [+ or -] 5 56 Sayala Planorbis 30.6 [+ or -] 4 40 Parameter ALT (Unites/ml) Treatments Mean [+ or -] Change SD % Control lab 41.5 [+ or -] 10 Bort-Said Kaar Biomphalaria 64.3 [+ or -] 24 55 El-Bahr Planorbis 50.6 [+ or -] 7 22 El-Khankak Biomphalaria 57.9 [+ or -] 0 40 Kobry Biomphalaria 74.8 [+ or -] 12 80 El-Lansh Planorbis 65.7 [+ or -] 11 58 Dakahlya Gammalya Biomphalaria 65 [+ or -] 15 57 Nasayma Biomphalaria 44.1 [+ or -] 10 6 Planorbis 85.8 [+ or -] 9 * 107 Physa 66.4 [+ or -] 10 60 Matarya Biomphalaria 44 [+ or -] 17 6 Damietta Annanya Physa 53.6 [+ or -] 7 29 Sayala Planorbis 51.2 [+ or -] 8 23 Parameter ALP (IU/L) Treatments Mean [+ or -] Change SD % Control lab 38.4 [+ or -] 8 Bort-Said Kaar Biomphalaria 42.5 [+ or -] 4 11 El-Bahr Planorbis 56.8 [+ or -] 11 48 El-Khankak Biomphalaria 106 [+ or -] 0.0 176 Kobry Biomphalaria 94.0 [+ or -] 29 145 El-Lansh Planorbis 101.4 [+ or -] 11 * 164 Dakahlya Gammalya Biomphalaria 65.5 [+ or -] 10 71 Nasayma Biomphalaria 52.0 [+ or -] 4 35 Planorbis 105.6 [+ or -] 10 * 175 Physa 162.3 [+ or -] 4.7 ** 323 Matarya Biomphalaria 66.3 [+ or -] 3.3 * 73 Damietta Annanya Physa 112.7 [+ or -] 4 ** 193 Sayala Planorbis 57.9 [+ or -] 11 51 Parameter Glucose Treatments Mean [+ or -] Change SD % Control lab 45.0 [+ or -] 8 Bort-Said Kaar Biomphalaria 76.1 [+ or -] 27 69 El-Bahr Planorbis 72.4 [+ or -] 11 61 El-Khankak Biomphalaria 85.7 [+ or -] 0.0 90 Kobry Biomphalaria 99.2 [+ or -] 6 * 120 El-Lansh Planorbis 96.8 [+ or -] 13 * 115 Dakahlya Gammalya Biomphalaria 91.0 [+ or -] 23 102 Nasayma Biomphalaria 80.1 [+ or -] 19 78 Planorbis 131.1 [+ or -] 6 ** 191 Physa 119.3 [+ or -] 9 * 165 Matarya Biomphalaria 85.3 [+ or -] 3.1 * 90 Damietta Annanya Physa 66.4 [+ or -] 12 48 Sayala Planorbis 67.4 [+ or -] 16 50 Parameter Creatinine (mg/dl) Treatments Mean [+ or -] Change SD % Control lab 0.55 [+ or -] 0.22 Bort-Said Kaar Biomphalaria 0.35 [+ or -] 0.0 -36 El-Bahr Planorbis 0 [+ or -] 0.0 -100 El-Khankak Biomphalaria 1.46 [+ or -] 0.0 165 Kobry Biomphalaria 0 [+ or -] 0.0 -100 El-Lansh Planorbis 1.53 [+ or -] 1.25 178 Dakahlya Gammalya Biomphalaria 0.40 [+ or -] 0.04 -27 Nasayma Biomphalaria 1.31 [+ or -] 0.94 138 Planorbis 0.38 [+ or -] 0.0 -31 Physa 1.41 [+ or -] 0.0 156 Matarya Biomphalaria 0.12 [+ or -] 0.0 -78 Damietta Annanya Physa 0.08 [+ or -] 0.0 -85 Sayala Planorbis 0 [+ or -] 0.0 -100 Parameter urea (mg/dl) Treatments Mean [+ or -] Change SD % Control lab 9.4 [+ or -] 1 Bort-Said Kaar Biomphalaria 13.8 [+ or -] 1 47 El-Bahr Planorbis 12.4 [+ or -] 0 32 El-Khankak Biomphalaria 30 [+ or -] 0.0 * 219 Kobry Biomphalaria 27.3 [+ or -] 1.2 ** 190 El-Lansh Planorbis 25.9 [+ or -] 1.1 ** 176 Dakahlya Gammalya Biomphalaria 22.9 [+ or -] 13 ** 144 Nasayma Biomphalaria 15.7 [+ or -] 3 67 Planorbis 19.3 [+ or -] 2 * 105 Physa 20.4 [+ or -] 2 * 117 Matarya Biomphalaria 22.8 [+ or -] 8 143 Damietta Annanya Physa 14.5 [+ or -] 3 54 Sayala Planorbis 12.3 [+ or -] 1 31 *, ** & *** significant compared to control value at p<0.05, p<0.01 & p<0.001, respectively. Table 2: Total protein, Albumin, globulin, A/G ratio, total Bilirubin, direct and indirect in tissue extract of snails collected from Lake Manzala. Parameter Total protein (g/dl) Treatments Mean [+ or -] SD Change % Control lab 6.21 [+ or -] 0.1 Bort- Kaar Biomphalaria 10.35 [+ or -] 0.4 ** 67 Said El-Bahr Planorbis 11.97 [+ or -] 0.6 ** 93 El- Biomphalaria 12.42 [+ or -] 0.04 ** 100 Khankak Kobry Biomphalaria 12.39 [+ or -] 0.8 ** 100 El- Planorbis 10.86 [+ or -] 0.6 ** 75 Lansh Dakahlya Gammalya Biomphalaria 11.49 [+ or -] 0.3 ** 85 Nasayma Biomphalaria 23.04 [+ or -] 0.5 *** 271 Planorbis 24.27 [+ or -] 0.7 *** 291 Physa 8.94 [+ or -] 0.2 ** 44 Matarya Biomphalaria 11.67 [+ or -] 0.59 ** 88 Damietta Annanya Physa 9.9 [+ or -] 0.6 * 59 Sayala Planorbis Parameter Albumin (g/dl) Treatments Mean [+ or -] SD Change % Control lab 3.27 [+ or -] 0.02 Bort- Kaar Biomphalaria 6.84 [+ or -] 0.30 ** 109 Said El-Bahr Planorbis 5.52 [+ or -] 0.57 * 69 El- Biomphalaria 7.53 [+ or -] 0.26 ** 130 Khankak Kobry Biomphalaria 6.42 [+ or -] 0.08 *** 96 El- Planorbis 6.24 [+ or -] 0.51 * 91 Lansh Dakahlya Gammalya Biomphalaria 6.72 [+ or -] 0.40 ** 106 Nasayma Biomphalaria 13.95 [+ or -] 0.30 *** 327 Planorbis 12.66 [+ or -] 0.04 *** 287 Physa 6.24 [+ or -] 0.04 *** 91 Matarya Biomphalaria 7.95 [+ or -] 0.35 ** 143 Damietta Annanya Physa 5.1 [+ or -] 0.41 * 56 Sayala Planorbis Parameter Globulin (g/dl) Treatments Mean [+ or -] SD Change % Control lab 2.94 [+ or -] 0.11 Bort- Kaar Biomphalaria 3.54 [+ or -] 0.11 * 20 Said El-Bahr Planorbis 6.45 [+ or -] 0.04 *** 119 El- Biomphalaria 4.89 [+ or -] 0.30 * 66 Khankak Kobry Biomphalaria 3.87 [+ or -] 0.74 32 El- Planorbis 4.62 [+ or -] 0.12 ** 57 Lansh Dakahlya Gammalya Biomphalaria 4.77 [+ or -] 0.12 ** 62 Nasayma Biomphalaria 9.12 [+ or -] 0.22 *** 210 Planorbis 11.61 [+ or -] 0.63 ** 295 Physa 2.7 [+ or -] 0.15 -8 Matarya Biomphalaria 3.72 [+ or -] 0.25 27 Damietta Annanya Physa 4.8 [+ or -] 0.16 ** 63 Sayala Planorbis Parameter A/G Ratio <> Treatments Control lab 1.13 Bort- Kaar Biomphalaria 1.93 Said El-Bahr Planorbis 0.85 El- Biomphalaria 1.63 Khankak Kobry Biomphalaria 1.22 El- Planorbis 1.34 Lansh Dakahlya Gammalya Biomphalaria 1.44 Nasayma Biomphalaria 1.53 Planorbis 1.12 Physa 2.35 Matarya Biomphalaria 2.16 Damietta Annanya Physa 1.05 Sayala Planorbis Parameter Total Bilirubin (Umol/l) Treatments Mean [+ or -] SD Change % Control lab 2.8 [+ or -] 0.1 Bort- Kaar Biomphalaria 4.4 [+ or -] 0.0 * 1.6 Said El-Bahr Planorbis 4.8 [+ or -] 0.8 2 El- Biomphalaria Khankak Kobry Biomphalaria 6.3 [+ or -] 0.3 ** 3.5 El- Planorbis 6.5 [+ or -] 1.2 * 3.7 Lansh Dakahlya Gammalya Biomphalaria 5.3 [+ or -] 0.1 ** 2.5 Nasayma Biomphalaria 4.5 [+ or -] 0.0 * 1.7 Planorbis 10.4 [+ or -] 0.0 ** 7.6 Physa 14.7 [+ or -] 1.8 * 11.9 Matarya Biomphalaria 5.0 [+ or -] 0.1 ** 2.2 Damietta Annanya Physa 5.7 [+ or -] 0.9 * 2.9 Sayala Planorbis 4.8 [+ or -] 0.8 2 Parameter Direct Bilirubin (mg/ dl) Treatments Mean [+ or -] SD Change % Control lab 0.2 [+ or -] 0.04 Bort- Kaar Biomphalaria 0.6 [+ or -] 0.0 200 Said El-Bahr Planorbis 0.1 [+ or -] 0.0 -50 El- Biomphalaria Khankak Kobry Biomphalaria 0.3 [+ or -] 0.0 50 El- Planorbis 1.1 [+ or -] 0.1 ** 450 Lansh Dakahlya Gammalya Biomphalaria 0.1 [+ or -] 0.4 -50 Nasayma Biomphalaria 2.5 [+ or -] 0.0 ** 1150 Planorbis 0.4 [+ or -] 0.0 100 Physa 1.8 [+ or -] 1.3 800 Matarya Biomphalaria 0.4 [+ or -] 1.5 100 Damietta Annanya Physa 2.1 [+ or -] 0.4 * 950 Sayala Planorbis 0.3 [+ or -] 0.2 50 Parameter In-Direct Bilirubin (U/mg) Treatments Mean [+ or -] SD Change % Control lab 2.5 [+ or -] 0.1 Bort- Kaar Biomphalaria 3.8 [+ or -] 0.0 * 52 Said El-Bahr Planorbis 4.7 [+ or -] 0.8 88 El- Biomphalaria Khankak Kobry Biomphalaria 6.0 [+ or -] 0.3 ** 140 El- Planorbis 5.4 [+ or -] 0.4 ** 116 Lansh Dakahlya Gammalya Biomphalaria 5.1 [+ or -] 0.2 ** 104 Nasayma Biomphalaria 2.0 [+ or -] 0.0 -20 Planorbis 10.0 [+ or -] 0.0 * 300 Physa 12.9 [+ or -] 0.6 ** 416 Matarya Biomphalaria 4.4 [+ or -] 0.2 ** 76 Damietta Annanya Physa 3.6 [+ or -] 1.3 44 Sayala Planorbis 4.5 [+ or -] 0.7 80 *, ** & *** significant compared to control value at p<0.05, p<0.01 & p<0.001, respectively. <> A/G = Ratio of albumin /globulin concentration Table 3: Catalase (CAT), glutathione-S-transferase (GST) and Gamma- glutamyl transpeptidase (GGT) in tissue extract of snails collected from Lake Manzala. Parameter CAT (Unites/g) Treatments Mean [+ or -] SD Change % Field collected snails Control lab 9.06 [+ or -] 0.18 Port- Kaar Biomphalaria 3.175 [+ or -] 0.37 ** -65 Said El-Bahr Planorbis 15.15 [+ or -] 1.9 * 67 Kobry Biomphalaria 3.96 [+ or -] 0.67 ** -56 El-Lansh Planorbis 7.59 [+ or -] 0.00 -16 Dakahlya Gammalya Biomphalaria 7.92 [+ or -] 1.59 -13 Planorbis 16.7 [+ or -] 0.00 * 84 Nasayma Biomphalaria 10.69 [+ or -] 1.31 18 Planorbis 1.36 [+ or -] 0.25 *** -85 Physa 0.89 [+ or -] 0.00 ** -90 Matarya Biomphalaria 7.19 [+ or -] 0.05 ** -21 Physa 18.15 [+ or -] 0.00 ** 100 Planorbis 15.52 [+ or -] 0.00 * 71 Damietta Annanya Physa 7.195 [+ or -] 0.54 * -21 Planorbis 25.785 [+ or -] 1.83 ** 185 Sayala Planorbis 17.31 [+ or -] 0.00 ** 91 Parameter GST (Unites/g) Treatments Mean [+ or -] SD Change % Field collected snails Control lab 3.07 [+ or -] 0.5 Port- Kaar Biomphalaria 1.52 [+ or -] 0.18 -50 Said El-Bahr Planorbis 5.60 [+ or -] 0.13 * 82 Kobry Biomphalaria 2.11 [+ or -] 0.26 -31 El-Lansh Planorbis 4.66 [+ or -] 0.0 52 Dakahlya Gammalya Biomphalaria 2.44 [+ or -] 0.33 -21 Planorbis 5.31 [+ or -] 0.0 73 Nasayma Biomphalaria 3.93 [+ or -] 0.49 28 Planorbis 0.83 [+ or -] 0.19 * -73 Physa 0.53 [+ or -] 0.0 -83 Matarya Biomphalaria 3.63 [+ or -] 0.72 18 Physa 6.63 [+ or -] 0.0 116 Planorbis 5.80 [+ or -] 0.0 89 Damietta Annanya Physa 4.04 [+ or -] 0.93 32 Planorbis 6.71 [+ or -] 0.41 * 119 Sayala Planorbis 3.46 [+ or -] 2.7 13 Parameter GGT (Unites/g) Treatments Mean [+ or -] SD Change % Field collected snails Control lab 999 [+ or -] 24 Port- Kaar Biomphalaria 653 [+ or -] 311 -35 Said El-Bahr Planorbis 160 [+ or -] 19 *** -84 Kobry Biomphalaria 1241 [+ or -] 172 24 El-Lansh Planorbis 1227 [+ or -] 0 * 23 Dakahlya Gammalya Biomphalaria 909 [+ or -] 147 -9 Planorbis 984 [+ or -] 0 -1 Nasayma Biomphalaria 716 [+ or -] 118 -28 Planorbis 1152 [+ or -] 237 15 Physa 857 [+ or -] 0 -14 Matarya Biomphalaria 1274 [+ or -] 46 * 28 Physa 915 [+ or -] 0 -8 Planorbis 1056 [+ or -] 0 6 Damietta Annanya Physa 7651 [+ or -] 651 ** 666 Planorbis 3901 [+ or -] 731 * 290 Sayala Planorbis 1123 [+ or -] 266 12 *, ** & *** significant compared to control value at p< 0.05, p<0.01 & p<0.001, respectively. Table 4: Hematologic parameters of snails collected from Lake Manzala. Parameters Total cell count Examined Mean [+ or -] SD Change % Control 3.2 [+ or -] 1.0 Port Kaar Biomphalaria 2.25 [+ or -] 0.7 * -30 Said El-Bahr Planorbis 1.2 [+ or -] 0.3 *** -63 Kobry Biomphalaria 2.35 [+ or -] 0.6 -27 El-Lansh Planorbis 0.95 [+ or -] 0.1 *** -70 Matarya Biomphalaria 2.6 [+ or -] 0.1 -19 Daka- Gammalya Biomphalaria 1.4 [+ or -] 0.0 *** -56 hlya Nasayma Biomphalaria 0.9 [+ or -] 0.4 *** -72 Sayala Planorbis 1.95 [+ or -] 0.0 ** -39 Damie Annanya Physa 2.65 [+ or -] 0.5 -17 tta Planorbis 1.63 [+ or -] 0.04 *** -49 Parameters Hyalinocytes Examined % Mean [+ or -] SD Change % Control 57 1.85 [+ or -] 0.5 Port Kaar Biomphalaria 61 1.4 [+ or -] 0.7 -24 Said El-Bahr Planorbis 57 0.7 [+ or -] 0.0 *** -62 Kobry Biomphalaria 60 1.35 [+ or -] 0.4 * -27 El-Lansh Planorbis 45 0.4 [+ or -] 0.0 *** -78 Matarya Biomphalaria 71 1.85 [+ or -] 0.2 0 Daka- Gammalya Biomphalaria 77 1.1 [+ or -] 0.0 *** -41 hlya Nasayma Biomphalaria Sayala Planorbis 70 1.4 [+ or -] 0.0 * -24 Damie Annanya Physa 70 1.85 [+ or -] 0.6 0 tta Planorbis Parameters Round small hemocytes Examined % Mean [+ or -] SD Change % Control 27 0.8 [+ or -] 0.28 Port Kaar Biomphalaria 24 0.5 [+ or -] 0.28 -38 Said El-Bahr Planorbis 24 0.25 [+ or -] 0.21 *** -69 Kobry Biomphalaria 24 0.55 [+ or -] 0.21 -31 El-Lansh Planorbis 41 0.35 [+ or -] 0.07 *** -56 Matarya Biomphalaria 18 0.45 [+ or -] 0.21 * -44 Daka- Gammalya Biomphalaria 16 0.2 [+ or -] 0.00 *** -75 hlya Nasayma Biomphalaria Sayala Planorbis 15 0.3 [+ or -] 0.0 *** -63 Damie Annanya Physa 23 0.55 [+ or -] 0.21 -31 tta Planorbis Parameters Granuolocytes Examined % Mean [+ or -] SD Change % Control 16 0.55 [+ or -] 0.2 Port Kaar Biomphalaria 15 0.35 [+ or -] 0.4 -36 Said El-Bahr Planorbis 16 0.25 [+ or -] 0.3 * -55 Kobry Biomphalaria 16 0.45 [+ or -] 0.5 -18 El-Lansh Planorbis 14 0.20 [+ or -] 0.0 *** -64 Matarya Biomphalaria 11 0.30 [+ or -] 0.3 -45 Daka- Gammalya Biomphalaria 7 0.10 [+ or -] 0.0 *** -82 hlya Nasayma Biomphalaria Sayala Planorbis 15 0.3 [+ or -] 0.0 ** -45 Damie Annanya Physa 7 0.25 [+ or -] 0.1 ** -55 tta Planorbis Parameters Hemoglobin (g/100ml) Examined Control 1.8 Port Kaar Biomphalaria 2.3 28 Said El-Bahr Planorbis 2.6 44 Kobry Biomphalaria 1.8 0 El-Lansh Planorbis 2.3 28 Matarya Biomphalaria 2.0 11 Daka- Gammalya Biomphalaria 1.7 -6 hlya Nasayma Biomphalaria Sayala Planorbis Damie Annanya Physa 0.8 -56 tta Planorbis *, ** & *** significant compared to control value at p<0.05, p<0.01 & p<0.001, respectively.
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|Title Annotation:||Research Article; Lake Manzala, Egypt|
|Author:||Khayat, Hanaa M.M. El-; Abdel-Hamid, Hoda; Gaber, Hanan S.; Mahmoud, Kadria M.A.; Flefel, Hassan E.|
|Publication:||Fisheries and Aquaculture Journal|
|Date:||Dec 1, 2015|
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