<?xml version="1.0" encoding="UTF-8"?><!DOCTYPE article  PUBLIC "-//NLM//DTD Journal Publishing DTD v3.0 20080202//EN" "http://dtd.nlm.nih.gov/publishing/3.0/journalpublishing3.dtd"><article xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" dtd-version="3.0" xml:lang="en" article-type="research article"><front><journal-meta><journal-id journal-id-type="publisher-id">AS</journal-id><journal-title-group><journal-title>Agricultural Sciences</journal-title></journal-title-group><issn pub-type="epub">2156-8553</issn><publisher><publisher-name>Scientific Research Publishing</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="doi">10.4236/as.2020.117040</article-id><article-id pub-id-type="publisher-id">AS-101788</article-id><article-categories><subj-group subj-group-type="heading"><subject>Articles</subject></subj-group><subj-group subj-group-type="Discipline-v2"><subject>Biomedical&amp;Life Sciences</subject><subject> Earth&amp;Environmental Sciences</subject></subj-group></article-categories><title-group><article-title>
 
 
  Chronic Exposure to No-Effect Concentration of Diazinon Induced Histological Lesions in Organs of &lt;i&gt;Clarias&lt;/i&gt; &lt;i&gt;gariepinus&lt;/i&gt;
 
</article-title></title-group><contrib-group><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Adekemi</surname><given-names>Florence Fagbohun</given-names></name><xref ref-type="aff" rid="aff1"><sup>1</sup></xref><xref ref-type="corresp" rid="cor1"><sup>*</sup></xref></contrib><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Olufunke</surname><given-names>Eunice Ola-Davies</given-names></name><xref ref-type="aff" rid="aff2"><sup>2</sup></xref></contrib><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Benjamin</surname><given-names>Obukowho Emikpe</given-names></name><xref ref-type="aff" rid="aff3"><sup>3</sup></xref></contrib><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Oluwafemi</surname><given-names>Obagbemiro</given-names></name><xref ref-type="aff" rid="aff4"><sup>4</sup></xref></contrib><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Olanike</surname><given-names>Kudirat Adeyemo</given-names></name><xref ref-type="aff" rid="aff4"><sup>4</sup></xref></contrib></contrib-group><aff id="aff2"><addr-line>2Department of Veterinary Physiology and Biochemistry, University of Ibadan, Ibadan, Nigeria</addr-line></aff><aff id="aff4"><addr-line>Fish and Wildlife Unit, Department of Veterinary Public Health and Preventive Medicine, University of Ibadan, Ibadan, Nigeria</addr-line></aff><aff id="aff3"><addr-line>Department of Veterinary Pathology, University of Ibadan, Ibadan, Nigeria</addr-line></aff><aff id="aff1"><addr-line>Department of Veterinary Laboratory Technology, Federal College of Animal Health &amp;amp; Production Technology, Moor Plantation,Ibadan, Nigeria</addr-line></aff><pub-date pub-type="epub"><day>08</day><month>07</month><year>2020</year></pub-date><volume>11</volume><issue>07</issue><fpage>627</fpage><lpage>637</lpage><history><date date-type="received"><day>11,</day>	<month>May</month>	<year>2020</year></date><date date-type="rev-recd"><day>25,</day>	<month>July</month>	<year>2020</year>	</date><date date-type="accepted"><day>28,</day>	<month>July</month>	<year>2020</year></date></history><permissions><copyright-statement>&#169; Copyright  2014 by authors and Scientific Research Publishing Inc. </copyright-statement><copyright-year>2014</copyright-year><license><license-p>This work is licensed under the Creative Commons Attribution International License (CC BY). http://creativecommons.org/licenses/by/4.0/</license-p></license></permissions><abstract><p>
 
 
  In all parts of the world pesticides have been found in the aquatic ecosystem and scientific evidence has also shown that they can enter the food chain. Diazinon is an organophosphate pesticide, widely used in agriculture to control a wide variety of suckling and leaf eating insects and recently in fish culture to suppress some parasitic diseases; nevertheless, there is little study on its adverse effect on fish. In this study, seventy-two (72) apparently healthy catfish comprising adult and juvenile of both sexes were used to set up triplicate experimental groups of those exposed to culture water alone (control group), fish exposed to pre-determined no-effect concentration (0.405 ppm) of Diazinon (test group). The fish were exposed for 28 days and fish were sacrificed and organs harvested on days 21 and 28 to determine the effect of long-term exposure to diazinon on organ histology. Histological changes observed in diazinon-exposed catfish were hyperplasia and fusion of the gill epithelium, hyperplasia of mucoid producing cells and aggregation of melanin pigment in the skin. Histological lesions were also seen observed in other organs, including severe diffuse cellular swelling and fatty degeneration of the liver, interstitial congestion of the kidney, carbon deposit on the wall of the heart and multifocal haemorrhage. The water quality of the control was not significantly different from that of the test group throughout the experiment. The lesions detected in cells, tissue, or organs represent an integration of cumulative effects of physiological and biochemical stressors. The histological alterations observed in vital organ of fish show that exposure to “no-effect” concentration of diazinon induced structural damage in fish organs and are likely to affect the functionality of the organs. For example, the adverse effect on the gill might disrupt its feeding and oxygen uptake.
 
</p></abstract><kwd-group><kwd>Diazinon</kwd><kwd> Histopathology</kwd><kwd> &lt;i&gt;Clarias&lt;/i&gt; &lt;i&gt;gariepinus&lt;/i&gt;</kwd></kwd-group></article-meta></front><body><sec id="s1"><title>1. Introduction</title><p>The end consequences of leaching and rain runoff, pesticides or pesticide residues are added continuously to the aquatic environment, thereby polluting fish and water bodies [<xref ref-type="bibr" rid="scirp.101788-ref1">1</xref>]. The natural aquatic systems may extensively be contaminated with indiscriminate use of pesticides, careless handling, accidental spillage, or discharges of untreated effluents into natural waterways and these will have harmful effects on fish population and other forms of aquatic life and may contribute long-term effects in the environment [<xref ref-type="bibr" rid="scirp.101788-ref2">2</xref>]. Water pollution by pesticides is a serious problem to all aquatic fauna, flora and to a considerable extent man [<xref ref-type="bibr" rid="scirp.101788-ref3">3</xref>]. Unfortunately, fish and water bodies are in grave danger because of bioaccumulation of pesticides. Many types of fish and different aquatic ecosystems have been adversely affected by pesticides which end up being suspended in water thereby making the water unfit for aquatic life [<xref ref-type="bibr" rid="scirp.101788-ref1">1</xref>].</p><p>In all parts of the world, pesticides have been found in the aquatic ecosystem and they also enter the food chain. Organophosphate pesticides are also used in fish culture in order to suppress some parasitic diseases nevertheless, the pesticide preparations are considered harmful to fish in most cases [<xref ref-type="bibr" rid="scirp.101788-ref4">4</xref>]. Fish are notorious for their ability to concentrate pollutants in their body tissues and since they play an important role in human nutrition, necessitating the need for screening to ensure the unduly high level of some toxic metals is not being transferred to man through fish consumption [<xref ref-type="bibr" rid="scirp.101788-ref5">5</xref>].</p><p>Clarias species is a widely distributed fish in Asia and Africa. In these areas, the fish is extremely popular on account of its tasty flesh, its unparalleled hardness, its rapid growth and high market price [<xref ref-type="bibr" rid="scirp.101788-ref6">6</xref>]. Clarias gariepinus is one of the choice fish distributed widely in Africa and other tropical countries of the world where they inhabit calm lakes, ponds, rivers and swamps in areas that are seasonally flooded. Catfish have inhabited all continents at one time or another [<xref ref-type="bibr" rid="scirp.101788-ref7">7</xref>] Due to the residual effects of pesticides, important organs are damaged. Acute and sublethal bioassays are carried out to check the short- and long-term effects of these organophosphates on animal life [<xref ref-type="bibr" rid="scirp.101788-ref8">8</xref>]. Among the common organophosphate used is Diazinon with the chemical formula (0,0-diethyl 0-[6-methyl-2(1-methylethyl)4-pyrimidinyl] is an organophosphate pesticide, is extensively used in agriculture and domestic pest control. This pesticide is also used to control a variety of insects including: aphids, beetles, scales and pill bugs, primarily in household environment and in agriculture crops [<xref ref-type="bibr" rid="scirp.101788-ref9">9</xref>].</p><p>The recent and notable work published on the effects of diazinon on various aspects of fish such as on the structure of the testis of bluegill [<xref ref-type="bibr" rid="scirp.101788-ref10">10</xref>]; on the common carp (Cyprinus carpio) embryos and larvae [<xref ref-type="bibr" rid="scirp.101788-ref11">11</xref>]; on expression and activity of intestinal P-glycoprotein [<xref ref-type="bibr" rid="scirp.101788-ref12">12</xref>]; on acetylcholinesterase activity and lipid peroxidation in the brain of Oreochromis niloticus. [<xref ref-type="bibr" rid="scirp.101788-ref13">13</xref>]; immunotoxicity and hepatic function evaluation in Nile tilapia (Oreochromis niloticus [<xref ref-type="bibr" rid="scirp.101788-ref14">14</xref>]; on acute toxicity of African catfish [<xref ref-type="bibr" rid="scirp.101788-ref15">15</xref>]; on the clinical and biochemical alterations associated with toxicity in Clarias gariepinus. [<xref ref-type="bibr" rid="scirp.101788-ref16">16</xref>]; on total protein and transaminase activities in Clarias gariepinus [<xref ref-type="bibr" rid="scirp.101788-ref17">17</xref>]; sublethal concentrations on total protein in tilapia fish (Oreochromis Niloticus) [<xref ref-type="bibr" rid="scirp.101788-ref18">18</xref>]; toxicological effect on African catfish (Clarias anguillaris) [<xref ref-type="bibr" rid="scirp.101788-ref19">19</xref>]; induced clastogenity and pathological changes in ovaries and testes of Clarias gariepinus [<xref ref-type="bibr" rid="scirp.101788-ref20">20</xref>] and on blood profile, histology of liver, gills and kidney of catfish, Clarias gariepinus [<xref ref-type="bibr" rid="scirp.101788-ref21">21</xref>].</p><p>It is imperative to find out the detrimental effects of pollutants specially pesticides on fish since they form an important link in food chain and their contamination by pesticides imbalance the aquatic system. Pesticides severely affect the fish in different ways mostly affecting the vital organs, therefore the present study was carried out to determine the histological changes of organs induced by chronic exposure of catfish (Clarias gariepinus) to diazinon.</p></sec><sec id="s2"><title>2. Methodology</title><p>Seventy-two (72) apparently healthy adult and juvenile catfish (Clarias gariepinus) of both sexes were set up in triplicate, adults and juveniles were exposed separately to avoid cannibalism. Four (4) catfish each (adult &amp; Juvenile) male and female were exposed to different treatment which includes pre-determined no effect concentration 0.405 ppm of diazinon [<xref ref-type="bibr" rid="scirp.101788-ref20">20</xref>] and water (control group). Route of exposure was via culture water. For each group, samples were collected on days 21 and 28 for histology which is the specify days for most chronic effects of fish in aquaculture (organs collected were skin, gills, kidney, hearts and liver) fixed in 10% neutral buffered formalin. Tissue specimens were processed routinely for paraffin sections of 4 - 5 μm thickness, stained with hematoxylin and eosin [<xref ref-type="bibr" rid="scirp.101788-ref22">22</xref>].</p></sec><sec id="s3"><title>3. Histopathology</title><p>For histopathological study, specimens from the skin, gills, kidneys, heart, and liver were collected from each fish on day 21 and 28, fixed in 10% neutral-buffered formalin. Tissue specimens were processed routinely for paraffin sections of 4 - 5 μm thickness, stained with hematoxylin and eosin [<xref ref-type="bibr" rid="scirp.101788-ref22">22</xref>]. The water quality like temperature, pH and dissolved oxygen were analyzed weekly 28.35˚C &#177; 0.75˚C, 7.50 &#177; 0.00 and 4.16 &#177; 0.28 mg/L respectively.</p></sec><sec id="s4"><title>4. Results</title></sec><sec id="s5"><title>5. Discussion</title><p>Fish and aquatic invertebrates have been considered to be an efficient and cost effective model system for studying the toxic, mutagenic, and carcinogenic potential of pollutants [<xref ref-type="bibr" rid="scirp.101788-ref23">23</xref>] [<xref ref-type="bibr" rid="scirp.101788-ref24">24</xref>] [<xref ref-type="bibr" rid="scirp.101788-ref25">25</xref>] [<xref ref-type="bibr" rid="scirp.101788-ref26">26</xref>] due to their ability to metabolise, concentrate, and store water-borne pollutants [<xref ref-type="bibr" rid="scirp.101788-ref27">27</xref>]. The exposure of fish to chemical contaminants induced a number of lesions in different organs [<xref ref-type="bibr" rid="scirp.101788-ref28">28</xref>]. Gills [<xref ref-type="bibr" rid="scirp.101788-ref29">29</xref>], kidney [<xref ref-type="bibr" rid="scirp.101788-ref30">30</xref>], and liver [<xref ref-type="bibr" rid="scirp.101788-ref31">31</xref>] are suitable organs for histological examination in order to determine the effect of pollution.</p><p>Histological changes when compared with the normal organs which was only exposed to cultured water made on skin, gills, kidneys, heart and liver of the diazinon exposed fish are represented in Figures 1-10. Showing hyperplasia of mucoid producing cells and aggregation of melanin pigment, Epithelial hyperplasia and fusion of the secondary epithelia, hyperplasia of hematopoietic cells and interstitial congestion, carbon deposits on the wall and severe diffuse cellular swelling and fatty degeneration showing by a massive hepatic degeneration respectively after 28 days’ expose to Diazinon respectively.</p><p>Gills and skin of fishes is the primary initial target of toxic chemicals, pesticides inclusive and cytological changes in gill morphology. Gills are mainly to be respiratory organs and any damage to them would cause disturbance in all physiological and metabolic activities by reduction in the supply of oxygen. It is quite evident that the damage caused to the gills results in impairments in gaseous exchange capacity of the gills, which ultimately lead to respiratory distress. The gills showing hyperplasia and fusion in this study, similar alterations in the gills have also been reported in the fishes exposed to metals [<xref ref-type="bibr" rid="scirp.101788-ref32">32</xref>] [<xref ref-type="bibr" rid="scirp.101788-ref33">33</xref>] [<xref ref-type="bibr" rid="scirp.101788-ref34">34</xref>] and organic contaminants [<xref ref-type="bibr" rid="scirp.101788-ref35">35</xref>] [<xref ref-type="bibr" rid="scirp.101788-ref36">36</xref>] [<xref ref-type="bibr" rid="scirp.101788-ref37">37</xref>] for the damage of gills in the fish treated with toxicants. The kidneys of fish exposed to diazinon showed histological changes after long time exposure of 28 days of treatment such as hyperplasia of hematopoietic cells and interstitial congestion. [<xref ref-type="bibr" rid="scirp.101788-ref38">38</xref>] has also reported that exposure of cypermethrin causes pathological effects on gills, liver and kidneys of catfish. Fusion of tubules and condensation of glomeruli content were reported by them. Also Pyknotic nucleus, destruction of fusion of tubules, condensation of nuclei was observed in the kidney of Clarias gariepinus exposed to diethyl phthalate [<xref ref-type="bibr" rid="scirp.101788-ref39">39</xref>] as seen in the present study.</p><p>The organ most associated with the detoxification and biotransformation process is the liver, and due to its function, position and blood supply [<xref ref-type="bibr" rid="scirp.101788-ref40">40</xref>]. Liver is an important organ for assessing the pollutant’s effects as the chemicals are stored in it. Severe diffuse cellular swelling and fatty degeneration of the liver (massive hepatic degeneration), hepatocytes necrosis may be the results of inability of the fish to regenerate new hepatocytes [<xref ref-type="bibr" rid="scirp.101788-ref37">37</xref>]. Necrosis occurred in the liver of exposed fish can also be related to the excessive burden of work for the detoxification of insecticide from the body. Similar line of reasoning for the liver necrosis was presented by [<xref ref-type="bibr" rid="scirp.101788-ref41">41</xref>] in fish treated with toxicant. [<xref ref-type="bibr" rid="scirp.101788-ref39">39</xref>] [<xref ref-type="bibr" rid="scirp.101788-ref42">42</xref>] [<xref ref-type="bibr" rid="scirp.101788-ref43">43</xref>] have reported necrosis of tubular epithelium and pyknotic nuclei in the liver of fish exposed to chemicals. In this study, the heart presented with carbon deposits on the wall which can also be indicative of reduction of oxygen for the circulation of blood to all part of system leading anaemia which can be injurious to the fish wellbeing.</p></sec><sec id="s6"><title>6. Conclusions</title><p>The histopathological changes observed in the skin, gills, kidney, heart and liver of the Clarias gariepinus in the present study indicate that the fish were responding to the direct effects of the contaminants. Such information confirms that histopathological alterations are good biomarkers for field assessment, in particular in tropical areas that are naturally subject to a multiplicity of environmental variations. As a result, this study showed histopathological changes in different organs of the catfish exposed to prolonged dose of diazinon, giving an affirmatory of diazinon having a detrimental effect on the cellular integrity of the organs. Therefore, in culturing, these could lead to poor performance, reduced productivity, compromised immune status and may eventually lead to death of the fish which at large can result in economic detune of the fish industry.</p><p>Therefore, strict and monitory control of the use of diazinon is hereby recommended to prevent harmful contact on feral and cultured fish.</p></sec><sec id="s7"><title>Conflicts of Interest</title><p>The authors declare no conflicts of interest regarding the publication of this paper.</p></sec><sec id="s8"><title>Cite this paper</title><p>Fagbohun, A.F., Ola-Davies, O.E., Emikpe, B.O., Obagbemiro, O. and Adeyemo, O.K. (2020) Chronic Exposure to No-Effect Concentration of Diazinon Induced Histological Lesions in Organs of Clarias gariepinus. Agricultural Sciences, 11, 627-637. https://doi.org/10.4236/as.2020.117040</p></sec></body><back><ref-list><title>References</title><ref id="scirp.101788-ref1"><label>1</label><mixed-citation publication-type="other" xlink:type="simple">Ahirwar, M.K., Shammi, Q.J., Chalko, S.R. and Lone, G.N. (2012) Effect of Malathion on Physio-Biological Aspects of Notopterus notopterus (Pallas). Asian Journal of Biological and Life sciences, 1, 159-163.</mixed-citation></ref><ref id="scirp.101788-ref2"><label>2</label><mixed-citation publication-type="other" xlink:type="simple">Ayoola, S.O. and Ajani, E.K. (2007) Histopathological Effects of Cypermethrinon Juvenile Nile Tilapia (Oreochromis niloticus) African Journal of Livestock Extension, 5, 1-13.</mixed-citation></ref><ref id="scirp.101788-ref3"><label>3</label><mixed-citation publication-type="other" xlink:type="simple">Ayoola S.O (2008) Histopathological Effects of Glyphosate on Juvenile African Catfish (Clarias gariepinus). Eurasian Journal of Agricultural and Environmental Science, 4, 362-367.</mixed-citation></ref><ref id="scirp.101788-ref4"><label>4</label><mixed-citation publication-type="other" xlink:type="simple">Robert and Hutson (1998) Metabolic Pathways of Agrochemical Parts 2; Insecticides and Fungicides. The Royal Society of Chemistry, Cambridge, 1475 p.</mixed-citation></ref><ref id="scirp.101788-ref5"><label>5</label><mixed-citation publication-type="other" xlink:type="simple">Ladipo, M.K., Doherty, V.F. and Oyebadejo, S.A. (2010) Acute Toxicity, Behavioural Changes and Histopathological Effect of Paraquat Dichloride on Tissues of Catfish (Clarias gariepinus). International Journal of Biology, 3, 67-74.https://doi.org/10.5539/ijb.v3n2p67</mixed-citation></ref><ref id="scirp.101788-ref6"><label>6</label><mixed-citation publication-type="other" xlink:type="simple">FAO (2003) Ecosystem Issues. OAR/National under Sea Research Programme. G.Mc. Fall. Aquaculture Newsletter-No. 29. http://www.fao.org/</mixed-citation></ref><ref id="scirp.101788-ref7"><label>7</label><mixed-citation publication-type="other" xlink:type="simple">Nelson, J.S. (2006) Fishes of the World. John Wiley &amp; Sons, Inc., Hoboken.</mixed-citation></ref><ref id="scirp.101788-ref8"><label>8</label><mixed-citation publication-type="other" xlink:type="simple">Sampath, K., Velamnial, S., Kennedy, I.J. and James, R. (1993) Haematological Changes and Their Recovery in Oreochromis mossambicus as a Function of Exposure Period and Sub-Lethal Levels of Ekalus. Acta Hydrobiologica, 35, 73-83.</mixed-citation></ref><ref id="scirp.101788-ref9"><label>9</label><mixed-citation publication-type="other" xlink:type="simple">Cong, N.V., Phuong, N.T. and Bayley, M. (2009) Effects of Repeated Exposure of Diazinon on Cholinesterase Activity and Growth in Snakehead Fish (Channa striatus). Ecotoxicology and Environmental Safety, 72, 699-703. https://doi.org/10.1016/j.ecoenv.2008.10.007</mixed-citation></ref><ref id="scirp.101788-ref10"><label>10</label><mixed-citation publication-type="other" xlink:type="simple">Dutta, H.M. and Meijer, H.J.M. (2003) Sublethal Effects of Diazinon on the Structure of the Testis of Bluegill, Lepomis macrochirus: A Microscopic Analysis. Environmental Pollution, 125, 355-360. https://doi.org/10.1016/S0269-7491(03)00123-4</mixed-citation></ref><ref id="scirp.101788-ref11"><label>11</label><mixed-citation publication-type="other" xlink:type="simple">Aydin, R. and K&amp;#246prücü, K. (2005) Acute Toxicity of Diazinon on the Common Carp (Cyprinus carpio L.) Embryos and Larvae. Pesticide Biochemistry and Physiology, 82, 220-225. https://doi.org/10.1016/j.pestbp.2005.03.001</mixed-citation></ref><ref id="scirp.101788-ref12"><label>12</label><mixed-citation publication-type="other" xlink:type="simple">Lecoeur, S., Videmann, B. and Mazallon, M. (2006) Effect of Organophosphate Pesticide Diazinon on Expression and Activity of Intestinal P-Glycoprotein. Toxicology Letters, 161, 200-209. https://doi.org/10.1016/j.toxlet.2005.09.003</mixed-citation></ref><ref id="scirp.101788-ref13"><label>13</label><mixed-citation publication-type="other" xlink:type="simple">Uner, N., Oru&amp;#231, E.&amp;#214., Sevgiler, Y., Sahin, N., Durmaz, H. and Usta, D. (2006) Effects of Diazinon on Acetylcholinesterase Activity and Lipid Peroxidation in the Brain of Oreochromis niloticus. Environmental Toxicology and Pharmacology, 21, 241-245. https://doi.org/10.1016/j.etap.2005.08.007</mixed-citation></ref><ref id="scirp.101788-ref14"><label>14</label><mixed-citation publication-type="other" xlink:type="simple">Girón-Pérez, M.I., Santerre, A., Gonzalez-Jaime, F., Casas-Solis, J., Hernández Coronado, M., Peregrina-Sandoval, J., Takemura, A. and Zaitseva, G. (2007) Immunotoxicity and Hepatic Function Evaluation in Nile Tilapia (Oreochromis niloticus). Exposed to Diazinon. Fish and Shellfish Immunology, 23, 760-769. https://doi.org/10.1016/j.fsi.2007.02.004</mixed-citation></ref><ref id="scirp.101788-ref15"><label>15</label><mixed-citation publication-type="other" xlink:type="simple">Adedeji, O.B., Adedeji, A.O., Adeyemo, O.K. and Agbede, S.A. (2008) Acute Toxicity of Diazinon to the African Catfish (Clarias gariepinus). African Journal of Biotechnology, 7, 651-654.</mixed-citation></ref><ref id="scirp.101788-ref16"><label>16</label><mixed-citation publication-type="other" xlink:type="simple">Bakhshwan, S., Hamed, H., Marzouk, M. and Hanna, M. (2009) Some Investigations on the Clinical and Biochemical Alterations Associated with Dizinon Toxicity in Clarias gariepinus. Egyptian. Journal of Aquatic Biology and Fisheries, 13, 173-197. https://doi.org/10.21608/ejabf.2009.2039</mixed-citation></ref><ref id="scirp.101788-ref17"><label>17</label><mixed-citation publication-type="other" xlink:type="simple">Inyang, I.R., Daka, E.R. and Ogamba, E.N. (2010) Effects of Sub-Lethal Concentrations of Diazinon on Total Protein and Transaminase Activities in Clarias gariepinus. Current Research Journal of Biological Sciences, 2, 390-395.</mixed-citation></ref><ref id="scirp.101788-ref18"><label>18</label><mixed-citation publication-type="other" xlink:type="simple">Soyingbe, A.A., Ogunyanwo, O.O., Hammed, T.B. and Adesope, A.O. (2012) Effects of Sublethal Concentrations of Diazinon on Total Protein in Tilapia Fish (Oreochromis niloticus). IOSR Journal of Environmental Science, Toxicology and Food Technology, 1, 22-25. https://doi.org/10.9790/2402-0112225</mixed-citation></ref><ref id="scirp.101788-ref19"><label>19</label><mixed-citation publication-type="other" xlink:type="simple">Adebayo, I.A., Akin-Obasola, B.J. and Bajulaye, O.M. (2013) Toxicological Effect of Diazinon on African Catfish (Clarias anguillaris). IOSR Journal of Environmental Science, Toxicology and Food Technology, 3, 64-71. https://doi.org/10.9790/2402-0316471</mixed-citation></ref><ref id="scirp.101788-ref20"><label>20</label><mixed-citation publication-type="other" xlink:type="simple">Ola-Davies, O.E., Fagbohun, A.F., Emikpe, B.O. and Adeyemo, O.K. (2015) Diazinon-Induced Clastogenity and Pathological Changes in Ovaries and Testes of Clarias gariepinus. Agricultural Science, 6, 146-151. https://doi.org/10.4236/as.2015.61012</mixed-citation></ref><ref id="scirp.101788-ref21"><label>21</label><mixed-citation publication-type="other" xlink:type="simple">Al-Otaibia, A.M., Al-Balawia, H.F.A., Ahmada, Z. and Sulimana. E.M. (2018) Toxicity Bioassay and Sub-Lethal Effects of Diazinon on Blood Profile and Histology of Liver, Gills and Kidney of Catfish, Clarias gariepinus. Brazilian Journal of Biology, 79. https://doi.org/10.1590/1519-6984.185408</mixed-citation></ref><ref id="scirp.101788-ref22"><label>22</label><mixed-citation publication-type="other" xlink:type="simple">Bancroft, J.D., Stevens, A. and Turner, D.R. (1996) Theory and Practice of Histological Techniques. 4th Edition, Churchill Livingstone, New York.</mixed-citation></ref><ref id="scirp.101788-ref23"><label>23</label><mixed-citation publication-type="other" xlink:type="simple">Belpaema, K., Delbeke, K., Zhu, L., Kirsch-Volders, M. (1996) Cytogenic Studies of PCB77 on Brown Trout (Salmo trutta fario) Using the Micronucleus Test and the Alkaline Comet Assay. Mutagenesis, 11, 485-492.https://doi.org/10.1093/mutage/11.5.485</mixed-citation></ref><ref id="scirp.101788-ref24"><label>24</label><mixed-citation publication-type="other" xlink:type="simple">Braunbeck, T., Boettcher, M., Hollert, H., Kosmehi, T., Lammer, E., Leist, E., Rudolf, M. and Seitz, N. (2005) Towards an Alternative for the Acute Fish LC (50) Test in Chemical Assessment: The Fish Embryo Toxicity Test Goes Multi-Species—An Update. Alternatives to Animal Experimentation, 22, 87-102.</mixed-citation></ref><ref id="scirp.101788-ref25"><label>25</label><mixed-citation publication-type="book" xlink:type="simple">Muller, W.E.G., Ed. (1994) Use of Aquatic Invertebrates as Tools for Monitoring of Environmental Hazards. Gustav Fischer Verlag, Stuttgart, 201-211.</mixed-citation></ref><ref id="scirp.101788-ref26"><label>26</label><mixed-citation publication-type="other" xlink:type="simple">Spitsbergen, J.M. and Kent, M.L. (2003) The State of the Art of the Zebrafish Model for Toxicology and Toxicologic Pathology Research—Advantages and Current Limitations. Toxicologic Pathology, 31, 62-87. https://doi.org/10.1080/01926230390174959</mixed-citation></ref><ref id="scirp.101788-ref27"><label>27</label><mixed-citation publication-type="other" xlink:type="simple">Osman, A.G.M., Wuertz, S., Mekkawy, I.A., Exner, H.J. and Kirschbaum, F. (2007) Lead Induced Malformations in Embryos of the African Catfish Clarias gariepinus (Burchell, 1822). Environmental Toxicology, 22, 375-389.https://doi.org/10.1002/tox.20272</mixed-citation></ref><ref id="scirp.101788-ref28"><label>28</label><mixed-citation publication-type="other" xlink:type="simple">Bucke, D., Vethaak, D., Lang, T. and Mellergaard, S. (1996) Common Diseases and Parasites of Fish in the North Atlantic: Training Guide for Identification. International Council for the Exploration of the Sea Techniques in Marine Environmental Sciences, Copenhagen.</mixed-citation></ref><ref id="scirp.101788-ref29"><label>29</label><mixed-citation publication-type="book" xlink:type="simple">Poleksic, V. and Mitrovic-Tutundzic. V. (1994) Fish Gills as a Monitor of Sublethal and Chronic Effects of Pollution. In: Müller, R. and Lloyd, R., Eds., Sublethal and Chronic Effects of Pollutants on Freshwater Fish, Fishing News Books, Oxford, 339-352.</mixed-citation></ref><ref id="scirp.101788-ref30"><label>30</label><mixed-citation publication-type="other" xlink:type="simple">Bucher, F. and Hofer, R. (1990) Effect of Domestic Wastewater on Serum Enzyme Activities of Brown Trout (Salmo truta). Comparative Biochemistry and Physiology, 97, 381-385. https://doi.org/10.1016/0742-8413(90)90158-6</mixed-citation></ref><ref id="scirp.101788-ref31"><label>31</label><mixed-citation publication-type="other" xlink:type="simple">ICES (International Council for the Exploration of the Sea) (1997) Report of Special Meeting on the Use of Liver Pathology of Flatfish For Monitoring Biological Effects of Contaminants. International Council for the Exploration of the Sea, Copenhagen.</mixed-citation></ref><ref id="scirp.101788-ref32"><label>32</label><mixed-citation publication-type="other" xlink:type="simple">Oliveira Ribeiro, C.A., Pelletier, E., Pfeiffer, W.C. and Rouleau, C. (2000) Comparative Uptake, Bioaccumulation, and Gill Damages of Inorganic Mercury in Tropical and Nordic Freshwater Fish. Environmental Research, 83, 286-292.https://doi.org/10.1006/enrs.2000.4056</mixed-citation></ref><ref id="scirp.101788-ref33"><label>33</label><mixed-citation publication-type="other" xlink:type="simple">Cerqueira, C.C.C. and Fernandes. M.N. (2002) Gill Tissue Recovery after Cooper Exposure and Blood Parameter Responses in the Tropical Fish Prochilodus scrofa. Ecotoxicology and Environmental Safety, 52, 83-91.https://doi.org/10.1006/eesa.2002.2164</mixed-citation></ref><ref id="scirp.101788-ref34"><label>34</label><mixed-citation publication-type="other" xlink:type="simple">Martinez, C.B.R., Nagae, M.Y., Zaia, C.T.B.V. and Zaia, D.A.M. (2004) Morphological and Physiological Acute Effects of Lead in the Neotropical Fish Prochilodus lineatus. Brazilian Journal of Biology, 64, 797-807.https://doi.org/10.1590/S1519-69842004000500009</mixed-citation></ref><ref id="scirp.101788-ref35"><label>35</label><mixed-citation publication-type="other" xlink:type="simple">Rosety-RodríGuez, M., Ordo&amp;#241ez, F.J., Rosety, M., Rosety, J.M., Ribelles, A. and Carrasco, C. (2002) Morpho-Histochemical Changes in the Gills of Turbot, Scophthalmus maximus L., Induced by Sodium Dodecyl Sulfate. Ecotoxicology and Environmental Safety, 51, 223-228. https://doi.org/10.1006/eesa.2001.2148</mixed-citation></ref><ref id="scirp.101788-ref36"><label>36</label><mixed-citation publication-type="other" xlink:type="simple">Fanta, E., Rios, F.S., Rom&amp;#227o, S., Vianna, A.C.C. and Freiberger, S. (2003) Histopathology of the Fish Corydoras paleatus Contaminated with Sublethal Levels of Organophosphorus in Water and Food. Ecotoxicology and Environmental Safety, 54, 119-130. https://doi.org/10.1016/S0147-6513(02)00044-1</mixed-citation></ref><ref id="scirp.101788-ref37"><label>37</label><mixed-citation publication-type="other" xlink:type="simple">Ezemonye, L. and Ogbomida, T.E. (2010) Histopathological Effects of Gammalin 20 on African Catfish (Clarias gariepinus). Applied Environmental and Soil Science, 2010, Article ID: 138019. https://doi.org/10.1155/2010/138019</mixed-citation></ref><ref id="scirp.101788-ref38"><label>38</label><mixed-citation publication-type="other" xlink:type="simple">Ayoola, S.O. and Ajani, E.K. (2008) Histopathological Effects of Cypermethrin on Juvenile African Catfish (Clarias gariepinus). World Journal of Biological Research, 1, 1-14.</mixed-citation></ref><ref id="scirp.101788-ref39"><label>39</label><mixed-citation publication-type="other" xlink:type="simple">Ikele, C.B., Mgbenka, B.O. and Oluah, N.S. (2011) Histopathological Effects of Diethyl Phthalate on Clarias gariepinus Juveniles. Animal Research International, 8, 1431-1438.</mixed-citation></ref><ref id="scirp.101788-ref40"><label>40</label><mixed-citation publication-type="other" xlink:type="simple">Van Der Oost, R., Beber, J. and Vermeulen, N.P.E. (2003) Fish Bioaccumulation and Biomarkers in Environmental Risk Assessment: A Review. Environmental Toxicology and Pharmacology, 13, 57-149.https://doi.org/10.1016/S1382-6689(02)00126-6</mixed-citation></ref><ref id="scirp.101788-ref41"><label>41</label><mixed-citation publication-type="other" xlink:type="simple">Rahman, M.Z., Hossain, Z., Mellah, M.F.R. and Ahmed, G.U. (2002) Effect of Diazinon 60EC on Anabus testudinus, Channa punctatus and Barbades gomonotus. Naga. The ICLARM Quarterly, 25, 8-11.</mixed-citation></ref><ref id="scirp.101788-ref42"><label>42</label><mixed-citation publication-type="other" xlink:type="simple">Biagianti-Risbourg, S. and Bastide, J. (1995) Hepatic Perturbation Induced by an Herbicide (Atrazine) in Juvenile Grey Mullet Liza ramada (Mugilidae, Teleotei): An Ultrastructural Study. Aquatic Toxicology, 31, 217-229. https://doi.org/10.1016/0166-445X(94)00065-X</mixed-citation></ref><ref id="scirp.101788-ref43"><label>43</label><mixed-citation publication-type="other" xlink:type="simple">Velmurugan, B., Selvanayagam, M., Cengiz, E.I. and Unlu, E. (2007) The Effect of Fenvalerate on Different Tissues of Freshwater Fish Cirrhinus mrigala. Journal of Environmental Science and Health, 42, 157-163.https://doi.org/10.1080/03601230601123292</mixed-citation></ref></ref-list></back></article>