June, 2020) P-ISSN: 1110-6344 Publisher: the Zoological Society of A

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Egyptian Journal of Zoology (EJZ), Vol. 73: 26-38 (June, 2020) p-ISSN: 1110-6344 Publisher: The Zoological Society of A. R. Egypt e-ISSN: 2682-3160 DOI: 10.12816/ejz.2020.25041.1022

RESEARCH ARTICLE

HEALTH STATUS OF DATE MUSSEL “LITHOPHAGA LITHOPHAGA, FAMILY: MYTILIDAE” IN ALEXANDERIA, EGYPT: POTENTIAL EFFECTS OF THE HEAVY METAL POLLUTION

Wesam M. Salama1*; Mohamed H. Mona1; Aml Z. Ghoneium2 1Zoology Department, Faculty of Science, Tanta University, Gharbia, Egypt 2Zoology Department, Faculty of Science, Damanhour University, Beheira, Egypt

ABSTRACT Article History: The date mussels (Lithophaga lithophaga) are rock-boring Received: 29 March 2020 bivalve molluscs that are generally used as bio-indicators for Revised: 9 April 2020 pollution. They can accumulate pollutants in their tissues, due to Accepted: 10 April 2020 their filter feeding behaviour. Therefore, the current study aimed Published Online: to use the date mussels to evaluate the heavy metal pollution 22 April 2020 in three coastal areas of Alexandria (Abo Qir, Alasafra, and Keywords: Miami) during the four seasons to determine the most polluted Bivalve molluscs area. The current study also aimed to evaluate the health status Heavy metals and disease burden of the date mussels collected from the Lithophaga lithophaga most polluted area according to the World Organisation for Pathology Seawater pollution Animal Health (OIE) manual. The results indicated that Pb, Cd, and Cu increased significantly in seawater of Abo Qir area *Correspondence: during summer, and was correlated with their accumulation in Wesam Salama the mussel tissues. As a consequence of pollution, a significant Zoology Department decrease in the amount of dissolved oxygen was observed in Faculty of Science Tanta University the seawater of Abo Qir area during summer. The pathological Gharbia, Egypt examination of samples from Abo Qir area using OIE E-mail: manual showed that the mussels had signs of weakness from wesam.hassan@science. the outer shell examination as gaping, fragility, discolouration, tanta.edu.eg and fouling. Moreover, infection with ciliates was observed in the date mussel tissues collected from the most polluted area. Interestingly, abnormal case of disseminated neoplasia or sarcoma also appeared under the effect of pollution. In conclusion, Abo Qir is the most polluted area with low dissolved oxygen concentration during summer. Moreover, accumulation of heavy metals in the soft parts of the date mussels could increase their disease susceptibility.

INTRODUCTION date mussels in Egypt. Heavy metals Date mussels (Lithophaga lithophaga may enter aquatic ecosystem from different Linnaeus, 1758) are bivalved Mollusca sources as natural and anthropogenic that considered one of the boring mussels on sources, which include industrial or any calcareous substrate[1]. Environmental domestic sewage[2]. Mussels are recognized variations and marine pollution may affect as pollution bio-indicator organisms because

26 Effect of pollution on health of date mussels in Egypt they accumulate pollutants in their soft MATERIAL AND METHODS tissues due to their filter feeding habits[3]. Studying area and sampling: Interestingly, bivalved Mollusca vulnerable Three locations were chosen from the to be infected with many pathogens as coastal regions of Alexandria (Abu Qir, viruses, fungi, bacteria, protozoa due Alasafra, and Miami, Figure 1) to study to heavy metals pollution that can the pollution with heavy metals during induce immune-suppression and reduction the four seasons in 2018. Water samples in disease resistance[4]. The interactions were seasonally collected in the morning between the two kinds of stresses as (5 replica/location/season) at depth of 3-5 m, infectious diseases and pollutants on aquatic filtered to remove any debris, and stored organisms may cause synergistic effect at 20°C until use. Molluscan samples that cause severe damage to a Molluscan “Lithophaga lithophaga” were collected population[5]. Protozoal infection considered seasonally from the three collection sites, as the major threats for farmed and natural pollution bio-indicators; whereas, samples bivalved species[6]. It is well known that were collected from the most polluted area heavy metals bioaccumulation and thermal during the significant season to evaluate of pollution have a major role in viral disease the disease burden of date mussels under the outbreaks in marine invertebrates[7]. Another effect of pollution. Samples were collected abnormal condition was noticed in Mollusca by scuba divers who break the rocks with exposed to pollution and infectious disease special sledge hammers without detrimental such as disseminated neoplasia or sarcomas; effect on the organisms living on their it defined as a proliferation of abnormal surfaces and within substratum. Collected circulating cells of unknown origin and samples (~50 samples/location/season) were defined as cancerous condition of bivalvia[8]. transferred within two hours in ice-box The current study aimed to evaluate the to central laboratory in Tanta University, heavy metals levels in seawater and their divided into groups, and labelled. Diameters bioaccumulation in soft parts of boring date or morphometric analysis of samples were mussels that collected from different coastal measured; and samples were dissected areas in Alexandria during the four seasons, for heavy metals analysis, biochemical as a bio-indicator for aquatic pollutant, and compositions, and disease detection using investigated the health status of date mussels World Organisation for Animal Health under the effect of pollution. (OIE) manual.

Figure 1: Google maps showing the collection sites along Alexandria coastal line.

27 Salama, W. M. et al.

Physicochemical properties of water (for transmission electron microscope, TEM). samples The fixed tissues with formalin were Surface water samples were collected five dehydrated and embedded in paraffin times from coastal regions of Alexandria wax. Then, thin sections were prepared, during the four seasons for measuring mounted, and stained using haematoxylin of pH, dissolved oxygen, temperature, and eosin (H & E) or toluidine blue. For and salinity. The value of pH was TEM, the fixed tissues were washed measured immediately after collection with phosphate buffer and dehydrated using Bench type pH meter (3510 Electro- with ascending grades of ethanol, and chemistry Analyser pH meter; Jenway, embedded in Epon 812 (Sigma-Aldrich Stone, UK). Dissolved oxygen was Chemie GmbH, Taufkirchen, Germany). determined according to the method of Grasshoff[9]. Temperature was measured Statistical analysis using an ordinary thermometer. Salinity All data are the means of five replicates. was determined by measuring the Normality of the data was tested with electrical conductivity using an inductive the Kolmogorov-Smirnov test. Two-way salinometer (model RS.10; Beckman, Cedar analysis of variance (ANOVA) was used Grove, NJ, USA). to test the effects of sites (3 levels) and seasons (4 levels) on the properties Heavy metal analysis of seawater sample and the heavy The heavy metals in both water and meats concentrations of seawater samples tissue samples were analysed according and soft tissues of date mussels‟ samples. to the method of Bernhard[10]. The heavy If there is a significant difference between metals of seawater samples were means, Tukey‟s post-hoc comparisons determined using Graphite Furnace Atomic among different groups were performed. Absorption Spectroscopy (model 2380; P-values <0.05 were considered to be Perkin-Elmer, Shelton, CT, USA). Soft parts statistically significant. from dissected samples were digested in Teflon vessels using concentrated HNCO3, dissolved in deionized distilled water, RESULTS and analysed using ion selective electrode Physicochemical parameters of seawater (AVL, 9180 electrolyte analyser, Roswell, samples GA, USA). Measurements of some physicochemical parameters of seawater samples from Pathological techniques listed by OIE the three studying area during the Date mussels “50 samples” were four seasons were shown in Table "1". collected from Abo Qir area (the most The results indicated that there are polluted area) in summer and examined a significant increase (P˂0.05) in pH according to OIE[11] to detect symptoms, of seawater samples collected from clinical aspects, and pathological conditions Abo Qir in summer and a significant using macroscopic, microscopic (histo- decrease in pH of seawater samples logical), and ultrastructural examinations. collected from Alasafra and Miami in In macroscopic or gross examination, winter. In addition, a significant decrease mussels were examined for any external (P˂0.05) in the amount of dissolved clinical abnormalities and clinical alterations. oxygen was recorded in seawater samples Parts of gills, mantle, gonads, and digestive collected from Abo Qir in almost all tract were removed from the dissected seasons, while it increased significantly samples and fixed in 10% neutral in seawater samples collected from buffered formalin (for histopathological Alasafra and Miami in almost all examination), or fixed in 2.5% glutar- seasons (4.0-4.6 mg/L, versus 5.7-5.9 and aldehyde solution and osmium tetroxide 5.0-5.7 mg/L, respectively).

28 Effect of pollution on health of date mussels in Egypt

Table 1: Physicochemical properties of seawater samples from the studying area during the four seasons.

Abo-Qir Al-Asafra Miami Sum Aut Win Spr Sum Aut Win Spr Sum Aut Win Spr 8.4 7.5 7.6 8.0 8.1 7.5 6.8 8.0 8.0 7.4 7.2 8.0 pH ±1.3 ±0.3 ±1.0 ±1.2 ±1.0 ±1.1 ±0.4 ±1.7 ±2.0 ±1.6 ±0.02 ±1.3 a ab ab ab ab ab b ab ab ab b ab

Dissolved 4.0 4.3 4.5 4.6 5.7 5.8 5.9 5.9 5.6 5.7 5.1 5.0 oxygen ±0.2 ±0.5 ±1.0 ±0.6 ±0.04 ±1.1 ±0.8 ±1.0 ±0.7 ±0.1 ±1.3 ±1.1 (mg/L) d d d cd ab ab a a ab ab ab bc

30.3 28.5 28.5 28.0 29.8 28.3 28.2 28.0 30.2 28.0 27.5 27.7 Temperature ±2.0 ±3.0 ±1.7 ±1.6 ±0.04 ±2.0 ±1.8 ±2.5 ±3.2 ±1.1 ±2.0 ±1.8 (°C) a c bc c ab bc c c a c c c

35.7 35.3 35.4 35.1 35.0 34.5 34.6 34.6 35.0 34.5 34.0 34.5 Salinity ±1.6 ±1.0 ±2.3 ±1.4 ±1.0 ±0.8 ±1.4 ±2.0 ±0.6 ±2.0 ±2.4 ±2.4 (ppt) a a a a a a a a a a a a Data are presented as means ± standard deviation (n=5). Sum: summer, Aut: autumn, Win: winter, Spr: spring, ppt: parts per thousand. Values with different letters in the same row were significantly different (P<0.05).

Seawater temperature was ranged a slight increase, but insignificant, in between 27.5 to 30.3◦C during the four the total weights compared with samples seasons in all stations. There are no collected from Abo Qir (data not shown). significant differences in salinity of seawater On the other hand, significant increases in all locations during the four seasons (P˂0.05) in the concentrations of “Pb, Cd, (Table 1). and Cu” were recorded in the samples Measurements of heavy metals in the collected from Abo Qir compared with the seawater samples from the three studying other sites during summer (Table 3). The areas indicated that the highest concentration relation between values of heavy metals of most heavy metals was found in in both seawater and soft parts of date seawater samples of Abo Qir during mussels changed significantly in most cases summer (Table 2). Obviously three of the according to the collection sites and seasons eight evaluated heavy metals (Pb, Cd, and (Table 4). Different collection sites and Cu) were significantly increased (P˂0.05) season could significantly (P˂0.05) affect in shallow seawater collected from Abo concentrations of Fe, Ni, Pb, and Co Qir during summer compared with Alasafra in seawater, and Ni, Pb, Cd, Cu, Co, and Cr and Miami shallow seawater at the same in soft parts of date mussels. season. Pathological examination of Abo Qir date Morphometric analysis and heavy metal mussels according to OIE concentrations of date mussels Macroscopic examination of 50 samples of There were no differences in shell heights, Abo Qir date mussels collected during total length, and width of date mussels summer showed that 33 samples (66%) collected from the three locations (data had external abnormal symptoms and not shown). While, samples collected from signs of weakness such as fouling, shell Alasafra and Miami in summer showed discolouration, shell fragility (Figure 2) and

29 Salama, W. M. et al.

Table 2: Heavy metals (g/L) in the seawater samples from the studying area during the four seasons.

Abo-Qir Alasafra Miami

Sum Aut Win Spr Sum Aut Win Spr Sum Aut Win Spr 6.5 6.3 5.0 5.5 5.9 5.6 5.4 5.2 4.2 3.9 3.5 3.8 Fe ±3.4 ±3.2 ±2.3 ±2.5 ±2.0 ±1.7 ±1.5 ±1.2 ±1.2 ±0.8 ±0.6 ±0.7 a ab bc bc ab ab ab abc cd cd d d 2.1 1.9 1.5 1.7 1.3 1.1 0.8 0.9 1.35 1.1 1.0 0.9 Ni ±1.0 ±0.8 ±0.6 ±0.7 ±0.4 ±0.2 ±0.1 ±0.2 ±0.1 ±0.1 ±0.1 ±0.1 ab a abcd abcd abcd bcd d d abcd bcd bcd cd 2.4 2.3 2.0 2.1 1.0 0.9 0.6 0.5 0.8 0.6 0.6 0.7 Pb ±1.5 ±1.2 ±0.9 ±1.0 ±0.6 ±0.5 ±0.4 ±0.3 ±0.2 ±0.1 ±0.1 ±0.1 a a a a b b b b b b b b 5.8 5.5 5.2 4.9 6.0 5.7 5.4 5.3 6.3 5.5 5.2 5.0 Zn ±2.1 ±2.0 ±1.8 ±1.5 ±0.1 ±0.1 ±0.04 ±0.02 ±1.0 ±0.8 ±0.6 ±0.5 ab ab b b ab ab ab ab a ab ab ab 2.8 2.6 2.1 2.4 1.0 0.9 0.6 0.7 0.3 0.2 0.1 0.1 Cd ±1.4 ±1.2 ±0.9 ±1.0 ±0.4 ±0.2 ±0.2 ±0.2 ±0.1 ±0.03 ±0.02 ±0.02 a a a a b bc bc bc bc c bc bc 2.5 2.3 2.2 2.0 1.6 1.4 1.2 1.7 1.4 1.1 0.9 1.0 Cu ±0.6 ±0.5 ±0.4 ±0.2 ±0.3 ±0.2 ±0.6 ±1.0 ±0.9 ±0.6 ±0.5 ±0.6 a a a ab bc bc c c c c c c 1.4 1.2 1.0 0.9 1.2 1.0 0.6 1.0 1.0 0.9 0.5 0.6 Co ±1.1 ±1.0 ±0.9 ±0.1 ±1.0 ±0.5 ±0.1 ±0.2 ±0.02 ±0.02 ±0.02 ±0.02 a ab ab abc ab ab bc ab ab abc c bc 1.6 1.4 1.2 1.0 1.4 1.1 0.9 0.6 1.0 1.0 1.6 1.8 Cr ±1.0 ±0.8 ±0.6 ±0.4 ±0.4 ±0.2 ±0.2 ±0.2 ±0.7 ±0.6 ±0.5 ±0.6 abc abc cd cd abc bc cd d cd cd ab a

Data are presented as means ± standard deviation (n=5). Sum: summer, Aut: autumn, Win: winter, Spr: spring. Values with different letters in the same row were significantly different (P<0.05).

gaping. Microscopic examination of mantle, Figures “3d and e”, where a transverse gills, gonad and digestive tract of date section of longitudinal bundle of micro- mussels collected from Abo Qir during tubules forming axoneme, which arranged summer using light and electron microscope into two arrays of microtubules 9 +2 arrays, showed that they were infected with was shown. Another abnormal condition Ciliophora. Haemocytes infiltration due to appeared in date mussels of polluted area heavy infection as an immune defence of was disseminated sarcomas or haemato- the sample was obvious in gills (Figures 3a). poietic neoplastic cells with hypertrophied Ciliates were characterized by its oval to enlarged cells than normal ones, which spherical shape and large macronucleus appeared in gills (Figure 3a) and digestive under light microscope, and appeared in tract as two nuclei cell (Figure 3f), or mussel gills (Figures 3a and b) and gonad with basophilic dense nuclei (Figure 3f (Figure 3c). The ultrastructure of ciliary and g) that had high nucleus: cytoplasm apparatus of Ciliophoran was clear in ratio.

30 Effect of pollution on health of date mussels in Egypt

Table 3: Heavy metals (mg/kg) in soft parts of collected Lithophaga lithophaga specimens from the studying area during the four seasons.

Abo-Qir Alasafra Miami Sum Aut Win Spr Sum Aut Win Spr Sum Aut Win Spr 5.4 5.2 5.0 5.3 4.1 4.8 4.5 4.2 4.6 3.9 3.5 3.7 Fe ±3.4 ±3.2 ±3.0 ±3.2 ±0.9 ±0.6 ±0.4 ±0.2 ±1.2 ±1.0 ±0.5 ±0.8 a ab ab ab bcd abc abc abcd abc cd d cd 0.9 0.7 0.6 0.4 0.6 0.4 0.3 0.4 1.0 0.8 0.7 0.8 Ni ±0.1 ±0.3 ±0.5 ±0.3 ±0.4 ±0.2 ±0.2 ±0.2 ±0.3 ±0.1 ±0.1 ±0.3 a ab bc c bc c c c a a ab a 2.1 1.9 2.0 1.7 0.9 0.9 0.5 0.5 0.8 0.6 0.6 0.6 Pb ±1.5 ±1.2 ±1.4 ±1.0 ±0.6 ±0.6 ±0.04 ±0.04 ±0.3 ±0.1 ±0.2 ±0.3 ab ab a b c cd e e cde de cde cde 5.3 5.0 4.9 4.9 5.0 4.5 4.4 4.5 4.9 4.5 4.2 4.5 Zn ±2.1 ±2.0 ±1.8 ±1.8 ±0.1 ±0.1 ±0.1 ±0.1 ±1.0 ±1.8 ±1.6 ±1.8 a a a a a a a a a a a a 2.4 0.9 0.8 1.0 1.4 2.2 1.8 2.0 1.5 1.2 0.9 1.0 Cd ±0.1 ±0.01 ±0.01 ±0.02 ±1.1 ±1.2 ±0.8 ±1.0 ±0.5 ±0.3 ±0.2 ±0.3 a cd d cd cd a ab ab bc cd cd cd 4.2 3.5 3.5 4.0 2.9 2.6 2.4 2.5 2.5 2.3 2.3 2.0 Cu ±0.9 ±0.6 ±0.6 ±0.8 ±1.0 ±0.9 ±0.5 ±0.7 ±0.3 ±0.2 ±0.3 ±0.02 a ab ab a bc cd cd cd cd cd cd d 0.9 0.8 0.5 0.5 0.8 0.7 0.6 0.6 0.6 0.5 0.5 0.5 Co ±0.5 ±0.4 ±0.02 ±0.1 ±0.3 ±0.1 ±0.04 ±0.1 ±0.02 ±0.02 ±0.1 ±0.01 a a a ab a ab ab ab b b b b 1.3 0.9 0.7 0.9 0.9 0.8 0.6 0.6 0.7 0.6 0.5 0.6 Cr ±1.0 ±0.1 ±0.6 ±0.1 ±0.5 ±0.4 ±0.3 ±0.2 ±0.3 ±0.2 ±0.1 ±0.1 a bcd cd bcd bcd bcd d d ab bcd bcd abc

Data are presented as means ± standard deviation (n=5). Sum: summer, Aut: autumn, Win: winter, Spr: spring. Values with different letters in the same row were significantly different (P<0.05).

DISCUSSION tissues depends on their concentration in Marine pollution has increased recently the environment, in addition to biological in the worldwide. It could affect the and environmental factors[2]. The deposition ecosystem and cause long term, negative of contaminants into a marine environment effects on the surrounding marine was possibly due to the transport of environment[12]. Pollution with heavy metals contaminants from upstream coastal in the aquatic environment increased industrial activities[14]. The present study recently under the effect of sewage showed that Pb, Cd, and Cu increased and industrial effluent[13]. Mussels are significantly in Abo Qir during summer recognized as a pollutant bio-indicator compared with the other coastal sites, because they accumulate high levels Miami and Alasafra. Radwan et al.[15] also of pollutant in their tissues from found that Abo Qir seawater mostly the surrounding marine environment[3]. The polluted with Cu and Cd compared with the bioaccumulation of heavy metals in mussels' seawater collected from Miami and El Mex.

31 Salama, W. M. et al.

Table 4: The relation between heavy metals, site of collection, and season in both seawater samples and soft parts of Lithophaga lithophaga samples.

Water samples Soft parts

F-value P-value F-value P-value Site of collection 76.51 P˂0.05 35.54 P˂0.05 Fe Season 9.96 P˂0.05 1.72 0.190

Site of collection 22.79 P˂0.05 68.11 P˂0.05 Ni Season 6.11 P˂0.05 11.52 P˂0.05

Site of collection 253.99 P˂0.05 514.67 P˂0.05 Pb Season 4.72 P˂0.05 11.30 P˂0.05

Site of collection 1.57 0.228 5.31 P˂0.05 Zn Season 7.32 P˂0.05 2.62 0.074

Site of collection 150.21 P˂0.05 35.19 P˂0.05 Cd Season 2.54 0.080 6.03 P˂0.05

Site of collection 59.03 P˂0.05 103.33 P˂0.05 Cu Season 0.87 0.471 5.26 P˂0.05

Site of collection 13.35 P˂0.05 47.86 P˂0.05 Co Season 11.15 P˂0.05 3.09 P˂0.05

Site of collection 8.30 P˂0.05 18.57 P˂0.05 Cr season 0.67 0.578 18.63 P˂0.05

a b

c d

Figure 2: Photomicrograph showing signs of weakness of Lithophaga lithophaga collected from Abo Qir. Their outer shell surfaces showed (a and b) fouling (black arrow indicates fouling of hydroid tube), (c) degrees of discolouration, (d) deformation and fragility (black arrow indicates holes and fragility).

32 Effect of pollution on health of date mussels in Egypt

a b c

d f g

Figure 3: Histological and ultrastructure of soft parts of Lithophaga lithophaga collected from Abo Qir area showing: (a) haemocytes infiltration (black arrows), neoplastic cells with pleomorphic nuclei (black arrow head) and ciliates (star) in gills of date mussel, bar = 100 µm, (b) ciliates embedded in gills, bar = 20 µm, (c) ciliates in gonad, bar = 20 µm, (d) ultrastructure of ciliary apparatus of ciliates in the digestive tract of mussel, bar = 100 nm, with axoneme arranged in two arrays of microtubules in transverse section (e), (f) neoplastic cells in digestive tract of date mussel showing cell with 2 nuclei (white arrow head) and basophilic nucleus (black arrow head), bar = 100 µm, (g) transmission electron microscopy showing cell with the basophilic nuclei (black arrow head), bar = 500 nm.

On the other hand, MacFarlane et al.[66] to agriculture, sewage, and industrial waste stated that Egyptian coastal area of effluents led to high concentration of Pb[18]. Mediterranean Sea might be considered As a consequence of water pollution in relatively unpolluted with heavy metals. the studying coastal areas of Alexandria The results of current study were evidenced in the current study, soft parts of date by the fact that, Abo-Qir station in mussels accumulate heavy metals in their Alexandria was polluted by the disposal tissue. Giustui et al. [19] confirmed that of effluents of both sewage and industrial marine organisms‟ tissues in polluted area activities. Ramadan et al.[67] found that could accumulate heavy metals. Bivalves Abo Qir station was affected by three could serve as pollution bio-indicators for discharges as Boughaz El-Maadia opening, metals in Egyptian water[20]. Fe is more Tapia pumping station and the opening of concentrated in date mussels‟ tissues Rashid Nile branch. The elevation of Cu followed by Zn> Cu >Cd >Pb> Cr> Co level in Abo Qir seawater shown in and Ni in the current study. Abdel Gawad[21] the current study might be due to leaching also detected the accumulation of Fe in of Cu from ships antifouling paints. Further- soft tissue of Lanistas carinatus that dwell more, Cd showed high concentration in in rock surfaces collected from Manzala the Abo Qir seawater in the present work Lake, Egypt. Freshwater Mollusca could during the four seasons. It was also found accumulate Fe, Cd, Pb, Mn, Cu, and Zn that leakage from ships painting, in addition in their tissues[22]. The ability of metal

33 Salama, W. M. et al. accumulation in soft parts differs from one and gonads of Abo Qir date mussels species to another according to rate of by TEM, as pear shaped, with large adsorption and digestion, or according to macronucleus and its cilia showed the pollution level, metabolic rate, and axonemes arrangement (9+2). In the north- age of the snail[22,23]. On the other hand, east coast of America, gill ciliates were pollution did not significantly affect also found in bivalvia collected from the protein, carbohydrates, moisture, fibre, and heavily polluted sites and the body ash contents, while lipid content somehow burden was correlated with the degree decreased, in the date mussels collected of pollutions[31]. Gill ciliates in mussels had from Abo Qir (data not shown). Other a negative correlation with Ni contamination, study reported a decrease in lipid contents but were positively correlated with Cd and of stressed aquatic organisms in polluted selenium contamination[32]. As the progress area due to consuming of energy during of sarcoma (neoplasia of mesoderm-derived eliminating the toxins from their bodies[24,25]. tissues), the haemocytes cells were replaced Heavy metals accumulation was affected by the neoplastic cells with pleomorphic by temperature, salinity, diet, and variations or basophilic nucleus, which unable to in marine organisms[26]. Water quality create the function of normal cells as depends on the adequate dissolved oxygen; phagocytosis and defence function, there- its ratio below 5 mg/L could cause stress fore the aquatic organisms become more to aquatic life[27]. The main causes susceptible to disease[33,34]. In the current of decreasing the dissolved oxygen study date mussels samples form Abo Qir concentration in seawater are the elevation showed neoplastic cells in digestive tract. in seawater temperature, algal bloom, animal The same finding was obtained by another and human wastes[27]. As a consequence study on Mediterranean mussels “Mytilus of pollution in Abo Qir during summer, galloprovincialis” collected from Slovene the seawater in this area showed the lowest Adriatic Sea[35]. Necrosis and multifocal value of dissolved oxygen (4.0±0.2 mg/L). atrophy of digestive tubules were noticed Pollutant and metals have a great in mussels with a diffuse neoplasia, whereas effect on the immune system of aquatic severe haemocytic infiltration of connective organisms and could increase susceptibility tissue was seen in mussels with single to infection[28]. Therefore, the bivalved neoplastic cells[35]. Possible cause of the Mollusca in polluted areas were vulnerable haemocytic neoplasia is a viral infection, but to infection with a lot of pathogens. In the other factors such as marine pollution and current study, there were no records for biotoxins may be involved[35]. Noticeably, infection with white spot syndrome, herpes- no viral infection was recorded in the present like, and irido-like viruses in date mussels study in the date mussels collected from the collected from Abo Qir (data not shown). most polluted area “Abo Qir” in summer that In contrary, viral pollution was recorded showed neoplastic cells in digestive tract; in different species of wild and farmed therefore, the heavy metals pollution may be bivalve molluscs in Alaska[29]. According the main cause for resulted disseminated to World Organization for Animal Health, sarcoma. Mollusca could be infected with protozoan In conclusion, the most polluted seawater parasites of genera Perkinsus, Bonamia, area with the heavy metals was the Abo Qir Martellia, and Haplosporidium that induced in summer in the current study. The mortality and caused enormous losses in seawater of this area was characterized by harvested molluscan species population[11]. a decrease in the amount of dissolve Prevalence could increase due to a lot of oxygen. In addition, Lithophaga lithophaga factors as high temperature in warm season, samples collected form Abo Qir showed high salinity, and pollution[30]. In the current accumulation of heavy metals (especially Pb, study, ciliates were easily defined in gills Cd, and Cu) and neoplastic cells in their

34 Effect of pollution on health of date mussels in Egypt soft parts, a decrease in immunity, and an and veterinary waste pharmaceutical increase in the susceptibility to ciliates exposure to wild aquatic host- infection parasite relationships. Environ Toxicol Pharmacol, 27(2): 161-175. [6] Fernández Robledo, J. A.; Vasta, G. R. ACKNOWLEDGMENTS and Record, N. R. (2014). Protozoan This research received no specific grant parasites of bivalve molluscs: literature from any funding agency in the public, follows culture. PLOS ONE, 9(6): commercial or not-for-profit sectors. e100872 (DOI: 10.1371/journal.pone.

0100872). CONFLICT OF INTEREST [7] Namroodi, S.; Milanlou, D. and The authors have no potential financial Rezaei, H. (2017). Heavy metal conflict of interest. bioaccumulation and its potential relation with incidence of canine parvovirus infection in golden AUTHORS’ CONTRIBUTIONS jackals, North Iran. Iran J Vet Med, WMS and MHM proposed the research idea, 11(2): 113-124. designed the experiments. WMS and AZG [8] Barber, B. J. (2004). Neoplastic performed the practical work, analysed diseases of commercially important the data, reviewed the literatures and data marine bivalves Aquat Living Resour, collection, as well as drafted the manuscript. 17: 449-466.

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How to cite this article: Salama, W. M.; Mona, M. H. and Ghoneium, A. Z. (2020). Health status of date mussel “Lithophaga Lithophaga, Family: Mytildae” in Alexanderia, Egypt: potential effects of the heavy metal pollution. Egyptian Journal of Zoology, 73: 26-38 (DOI: 10.12816/ejz.2020.25041.1022).

37 الوجلة الوصرية لعلن الحيىاى الوجلد 37 )يىًيى 0202(

الحالة الصحية لبلح البحر "٬Lithophaga lithophaga عائلة Mytilidae" في اإلسكٌدرية٬ جوهىرية هصر العربية: التأثيرات الوحتولة للتلىث بالوعادى الثقيلة

وسام هحود سالهه٬1 هحود حسي هٌا ٬1 أهل ذكي غٌين0 1 قسى عهى انحٕٛاٌ، كهٛخ انعهٕو، جبيعخ طُطب، انغشثٛخ، جًٕٓسٚخ يظش انعشثٛخ 0 قسى عهى انحٕٛاٌ، كهٛخ انعهٕو، جبيعخ ديُٕٓس، انجحٛشح، جًٕٓسٚخ يظش انعشثٛخ

ثهح انجحش "Lithophaga lithophaga" ْٕ أحذ إَٔاع انشخٕٚبد راد انًظشعٍٛ صبقجبد انظخٕس ٔانز٘ ٚسزخذو كًؤشش حٕٛ٘ نهزهٕس ثسجت رشاكى انًهٕصبد فٙ أَسجزّ انشخٕح حٛش أَّ ٚزغزٖ ثبنزششٛح. ْٔذفذ انذساسخ انحبنٛخ إنٗ اسزخذاو ثهح انجحش نزقٛٛى رهٕس يٛبِ انجحش ثبنًعبدٌ انضقٛهخ خالل األسثعخ يٕاسى فٙ صالس يُبطق سبحهٛخ ثبألسكُذسٚخ )أثٕ قٛش، ٔانعظبفشح، ٔيٛبيٙ( نزحذٚذ انًُطقخ األكضش رهٕصًب، ٔأٚضب رقٛٛى انحبنخ انظحٛخ نجهح انجحش ٔاكزشبف اإلطبثخ ثبأليشاع انًخزهفخ رحذ رأصٛش رهٕس انًٛبِ فٙ أكضش انًُبطق رهٕصب. ٔأٔضحذ انُزبئج أٌ رشكٛض كم يٍ انشطبص ٔانكبديٕٛو ٔانُحبط صادد ثشكم يهحٕظ فٙ يٛبِ انجحش ثًُطقخ أثٕ قٛش خالل فظم انظٛف، يًب أد٘ إنٙ رشاكًٓب فٙ األَسجخ انشخٕح نجهح انجحش. ٔكُزٛجخ نهًهٕصبد، نٕحع أٚضب اَخفبع كجٛش فٙ كًٛخ األكسجٍٛ انًزاة فٙ يٛبِ انجحش ثًُطقخ أثٕ قٛش خالل فظم انظٛف. ٔأظٓش فحض انعُٛبد انزٙ رى رجًٛعٓب يٍ يُطقخ أثٕ قٛش ثبسزخذاو دنٛم انًُظًخ انعبنًٛخ نظحّ انحٕٛاٌ "OIE" أٌ ثهح انجحش ٚعبَٙ أعشاع يشضّٛ يضم انفجٕاد، ٔانٓشبشخ، ٔرغٛش نٌٕ انسطح انخبسجٙ نهظذفخ. عالٔح عهٗ رنك، سُجهذ اإلطبثخ ثبنٓذثٛبد فٙ أَسجخ عُٛبد ثهح انجحش انزٙ رى جًعٓب يٍ أكضش انًُبطق رهٕصًب. ٔيٍ انًضٛش نالْزًبو ظٕٓس خالٚب غٛش طجٛعٛخ رعشف ثبنخالٚب انٕسيٛخ أٔ انسبسكٕيب فٙ أَسجخ ثهح انجحش ثسجت انزهٕس انجٛئٙ. َسزُزج يٍ رنك أٌ أثٕ قٛش يٍ أكضش انًُبطق رهٕصب ٔخظٕطب فٙ فظم انظٛف، يًب أدٖ إنٗ رشاكى انًهٕصبد فٙ أَسجخ ثهح انجحش ٔصٚبدح قبثهٛزّ نإلطبثخ ثبأليشاع.