Journal of Hematology & Thromboembolic Diseases
Open Access
ISSN: 2329-8790
ISSN: 2329-8790
Research Article - (2022)
Alterations in the blood components of the fresh water fish Labeo rohita are studied after exposing them to both technical grade and 10% EC of bifenthrin a synthetic pyrethroid as toxicants for 10 days in 1/10th of LC50 values 96 hrs LC50 values as sub lethal and lethal of 4 days. The parameters that are studied include Erythrocytes (RBC), Leucocytes (WBC), Haemoglobin (Hb) Haemotocrit (Ht), Mean Corpscular Volume (MCV), Mean Corpuscular Haemoglobin (MCH) and Mean Corpscular Haemoglobin Concentration (MCHC). The first four are determined and the last three are calculated values. Except WBC, all others are decreased as quantitative alterations and will be the impediments for the normal functioning of all physiological processes and are the one of the biomarker study of the toxicity and will be the indices of toxicity and pollution load. They are the one of the reasons for causing the death of the fish. The results are going to be discussed with earlier studies and objective is to not to allow the pollution of such caused by pesticides and in the agriculture practices and such contaminated waters should not be used as it is going to be detrimental for the impairment of growth for the fish which are cultured and the present studied one is also such that.
Bifenthrin; Technical grade 10% EC; Haematological studies; RBC; WBC; Hb; Hct (PCV); MCV; MCH and MCHC; Labeo rohita
Blood, is the nutritive, excretory, immunity media apart from the respiratory gases to carry for the living organisms especially more so in vertebrate group with a closed circulatory system. Metabolism of such heterotrophic organism including the fish depends on the quantitative components of its constituents of the mesoderm derivative which is differentiated into plasma and formed elements (Erythrocytes, Leucocytes and Platelets). In normal condition, fish health is dependent mainly on the circulatory fluid as it helps in proper metabolism carrying oxygen to different tissues/cells and also only defense mechanism which it provides with attacking by parasites or antigens or even in toxic stress [1].
The highly developed, vertebrates due to blood of its ecological adaptation are divided into exo and endoderm’s and also poikilotherms and homeotherms [2]. The fish candidate species for testing in the laboratory to offer any indices of the pollution due to the toxicant, pollutants, the pesticides as of one major category Fish are poikilothermic with a single circulation of blood of the various heart perceive any subtle changes in the waters with a sensory system. Any such situation of the, which external environment will be reflected as an effect in the blood, there by any components of its parameters quantitatively change had an impact on the overall survival of the organism/fish [3].
Bifenthrin, the toxicant selected for the study is resistant in freshwaters because it can have the process of break down neither can undergo the process of hydrolysis or photolysis or it can have metabolic nor in quick time [4]. It was a persistent one and also its bio concentration is also high because it is lipophilic and magnify in the food chain as per the same report as above. Hence, the best management practices to protect water and fish are 0.1 microgram/L which is the concentration that is permissible as a protector to the aquatic life and is considered as the ‘benchmark’ [5]. Hence, there is a justification in selecting the synthetic pyrethroid of type I, non-cyanogroup member the bifenthrin against a candidate species Labeo rohita one of the major carp and also a capture and culture fishery component as a part is selected as test species [6].
The studies of the haematological nature of the effects caused by the synthetic pyrethroids, the biomarker studies were extensively discussed and promoted by the review article by and also by [7]. They mentioned some of earlier reports exclusively for the pyrethroids including the present tested toxicant for the different species and also even in the review articles by; and also by which had a mention of the haematological alterations of the quantitative compounds of studies that can be the indices of pollution of the pesticides [8].
Hence, an attempt is made to study the variations of the quantitative nature in the blood components in the fish Labeo rohita both in lethal and sub lethal concentration of the bifenthrin a synthetic pyrethroid of type I, of its technical grade as well as its commercial formulation 10% EC, in the laboratory by following standard protocols [9].
Collection and maintenance of test organism
The freshwater fish Labeo rohita size and weight selected as per the toxicity experiments (3-5 cm length and 3-5 gms weight). Healthy and active fish were obtained from local fish farm of Nandivelugu, Guntur (AP), and India [10]. The fish were acclimatized to the laboratory conditions in large plastic water tanks for three weeks at a room temperature of 28 ± 1°C. Water was renewed every day with 12-12 h dark and light cycle. During the period of acclimatization, the fish were fed (ad libitum) with groundnut oil cake and rice bran. The feeding was stopped one day prior to the acute toxicity tests and also the hematological studies [11]. As per the recommendations of the precautions laid by the committee on toxicity tests to aquatic organism were followed [12]. The acclimated fish were selected for hematological evaluation. If the mortality, if any exceeded 5% in any batch of fish during acclimatization, the entire batch of that fish were discarded.
The water that was used for experimentation is similar to the one used for toxicity determination and the physical and chemical characteristics are [13].
Turbidity-8 silica units. Electrical conductivity at 28°C-816 Micro ohms/cm, pH at 28°C-8.1. Alkalinity Phenolphthalein=Nil, Alkalinity; Methyl orange-472, Total Hardness (as CACO3)-232, Carbonate Hardness (as CaCO3)-232, Non-Carbonate Hardness (as CaCO3)-Nil. Calcium Hardness (as CaCO3)-52, Magnesium Hardness-40, Nitrite Nitrogen (as N)-Nil, Sulphate (as SO42)-Trace Chloride (as Cl)-40; Fluoride (as F)-1.8, Iron (as Fe)-Ni, Dissolved Oxygen-8-10 ppm, Temperature-28 ± 2°C [14].
The permethrin technical grade is supplied by M/s. Kalyani Industries (Agrochemical suppliers 1202/1204, 12th floor, Bwing, Kailash Business Park, Ghotpur Power, India) and 10% EC is purchased in the local market.
The fish, fifty numbers each were exposed to technical grade and 10% EC in lethal and sub lethal concentrations for 4 days and the days respectively of 96 hrs concentration and the values are 0.22 μg/L and 0.022 μg/L (1/10th of LC50 value) and 0.11 μg/ L and 0.011 μg/L respectively for technical grade and 10% EC, as per recommendations of APHA (1998, 2005 and 2012 and OECD, 2019). A control of the fish without an exposure to the toxicants is also maintained a control. At the end of the exposure to fish are subjected to analysis of the blood constituents for observation of any haematological changes as alterations [15].
The Sampling of blood
Fish were euthanized by an overdose of MS-222 and then weighed and measured. Blood was sampled by caudal severance from the disease free test fish during the early hours of the day and stabilized with 50 IU sodium heparin (anticoagulant)/ml blood [16].
Haematological examination
The haematological variables analyzed by the standard procedures are RBC count, Haemoglobin (Hb), White Blood Cells Count (WBC), Haematocrit (Ht), Mean Corpuscular Volume (MCV), Mean Corpuscular Haemoglobin (MCH) and Mean Corpuscular Haemoglobin Concentration (MCHC) [17].
The RBC was determined by with a Nenbauer Crystalline Country Chouscher as described. The white blood cells count was determined by the method described. The Haemoglobin was estimated by cyanomethanoglobin method as described. The Mean Corpuscular Volume (MCV) was calculated by the following formula and expressed as fembliter [18].
MCV= Haematocrit % x 10/RBC count
The Mean Corpuscular Hemoglobin (MCH) was calculated by the following formula and expressed in pictogram (Pg) MCH=Hemoglobin (8./dL0 x 10/RBC count).
The Mean Corpuscular Hemoglobin Concentration (MCHC) was obtained by the following formula and express in terms of gram percent (8%).
The results after exposing the fish Labeo rohita (96 h) and sub lethal (1/10 of LC50 value for 10 days) concentrations bifenthrin is appreciable. The blood parameters estimated are presented graphically as (Figures 1 and 2) for technical grade and 10% EC in lethal concentrations and sub lethal and percent changes of hematological alterations in Labeo rohita on exposure to both sub lethal and 10% EC. Except in WBC all other parameters decreased both in lethal and sub lethal concentrations of technical grade as well as 10%. The impact of the commercial formulation in the alteration of parameters is significant and increase in WBC also [19].
Figure 1: Haematological changes in the parameters of blood of the fish Labeo rohita exposed to sub lethal and lethal concentrations of both Technical grade Bifenthrin as well as 10% EC.
Figure 2: Haematological changes of percentages in the parameters of blood of the fish Labeo rohita exposed in to sub lethal and lethal concentrations of both Technical grade Bifenthrin as well as 10% EC.
A decrease of RBC 19% (18.9%) in technical sub lethal and 23% (23.24%) concentration of technical grade when compared with 10% EC of sub lethal 7% and 11% in lethal showed, the decrement of the oxygen carrying capacity and fish is deprived of it.
Similarly the WBC count showed for technical grade sub lethal as 30.7% and 37.12% in lethal whereas for 10% EC of sub lethal as 3.4% and 8.71% in lethal as increase which can be presumed of that much increase an unspecific immunity due to the toxic stress to the technical grade toxic stress [20].
Similarly, the hemoglobin decreased in sub lethal by 35.62%and in lethal 43.75% whereas for 10% it is 6.25% and 11.25%and that much decrease of hemoglobin on oxygen carrying capacity and ultimately it has a bearing on Hematocrit values that too are changed by 26.17% and 19.61% for technical grade in lethal and sub lethal respectively, whereas for 10% EC it was 2.82% and 9.01% in sub lethal and lethal respectively.
The calculated values of MCV decreased lethal technical >sub lethal technical >lethal 10% EC >sub lethal >10% EC. Similarly MCH values decreased as similar of the above and MCHC too followed the same pattern.
Such appreciable changes in the blood parameters of the fish due to the toxic stress made a point clear that the fish in lethal concentration (acute), suffer the death whereas in sub lethal concentration (chronic) suffer in the long run too.
With group I and II synthetic pyrethroids
Sana Ullah, et al. in the fish Ctenopharyngodon idella using the bifenthrin as toxicant reported the changes in the blood components. Erythrolysis or inhibition of RBC production that was responsible for decrease in RBC count reflected on hemoglobin and also on hematocrit values. For the increase of WBC, lymphopoietin was responsible due to the toxic stress which is similar even to the present study.
Suchiang in his review article, specifically mentioned about the hematological aspects of changes of the pesticides in catfishes only. The reasons for decrement of RBC was due to exposure of the pesticides, in cat fishes Clarias gariepinus was due to direct ‘feedback’ mechanism which was also due to structural damage to RBC membrane, hemolysis or might be due to the impairment of its synthesis. The kidney is the hemopoietic tissue and the blood cells reservoir. Any decrement of the cells of it was due to dysfunction of the entire hemopoietic system of the fish. The increase of WBC due to an increased antibody production to cope the stress, which in a nutshell put it as disruption of the hematopoiesis and a decrease in the immunity of non-specific nature.
Saha and Saha in the fish Clarias batracus in the fish Heteropneustes fossils (Bloch). In the fish Oreochromis mossambicus too reported that the toxic stress manifested alterations as effects via the behavioral changes and the said parameters that showed variations. Even in the fish Oncorhynchus mykiss (rainbow trout) the bifenthrin the present tested toxicant had an impact on the fish.
Aysel opined that the pesticides at the concentration of the chronic levels due to oxidative stress, several biochemical alterations and in the hematological variations in different fishes.
Siebel, the fish blood gives much information about its physiological state of it. The contamination of the external medium is inevitable at any time and was perceived through the blood examination. Any such deviation of its quantitative internal components speaks about its ill-health or due to the effect of chemical stress.
Faraq, et al. in the fish Oreochromis niloticus due to bifenthrin intoxication resulted some neuronal behavioural and physiological alterations in which hematological changes are one of them.
Bano, et al. due to the effect of bifenthrin and chlorpyrifos in the fish Labeo rohita due to the toxic action reported that the erythrocytes were effected which resulted a decrement of them quantitatively which ultimately lead to impairment of the whole physiological respiratory process due to the reduction in the oxygen carrying capacity.
Deshmukh, et al. while investigating the polluted waters of river Godavari (near Paithan, Maharashtra state) in the fishes, Catla catla and Labeo rohita reported on the hematological changes due to contamination of the waters including the pesticides and of such act the blood of the fish showed variations which in the environment at the sub lethal levels in which such changes are quite possible.
Satyanarayana, et al. while reporting on the permethrin intoxication in the fish Ctenopharyngodon idella, the blood of the grass carp had changes/alterations. The results of the toxicant which also belongs to the same group of the present toxicant bifenthrin were similar (All parameters decreased except WBC), in both technical grade as well as 25% EC formulation exposures. Kidney damage by which blood regeneration is effected and due to decrement of the RBC and Histopathologically lesions of the gills had an impact on the oxygen carrying capacity and there was a decrement of the hemoglobin. The present tested fish Labeo rohita along with other two major carps and also the grass carp are cultured in polyculture practices and when such situation of the report who visualized the contaminated waters should not be used as a source for such aquacultures ventures.
In their review article gave a vivid aspects of the makers due to the pesticides toxic action to some of the different types of fish. The alterations of the blood such study Ullah Sana (2019b) reported on the changes in the fish Hypophthalmichus maltrix (Silver carp) of due to the deltamethrin type II with cyan group. Even Kartas (2016) in the fish Salmo trutta fario, RBC increased and decrement in other cells due to the same toxicant.
Exposed to the pesticides Lamba cyhalothrin a synthetic parathyroid of group I and domethoate an organophosphate in the fish Barbonymus gonionotus (Silver barb). They reported all the parameters decreased except MCHC (increased), in the blood of the fish exposed at two different concentrations of both toxicants and exposure time was only 96 hrs. The results are contradictory of the present study and even through the reasons are not mentioned in the report except of comparing the results of the earlier studies.
Virinalis (red swamp cray fish) and also to other cray fishes and all of them had an impact on the haematology. Except the oxygen carrying pigment (copper) and having closed system of blood circulations the shrimps that belong to the arthropods are similar to the fishes (poikilothermic/exothermic).
In the fish Heteropneustes fossils when exposed to waters contaminated with pesticides in Muzaffnagar (M.P.) had an impact on the blood parameters, which serve as indices of the pollution load especially with pesticides. Such contaminated waters should not be used as source for the culture due to the synergistic effects the variations that were reported are different for the different pesticides individually. The concentrations were at both lethal and sub lethal and were field/environmental study.
Reported on lambda cyhalothrin on the hematological alterations in the same fish which was selected for the present study, both in lethal as well as sub lethal concentrations of 24,48,72 and 96 hrs and 10, 20 and 40 days respectively. Only three parameters were studied RBC, WBC and Hg and the results that were obtained as of the present study. The toxicant they tested and the present studied one both belong to the same group of the synthetic parathyroid (type/group I non-cyano one). Stress factor had an impact and the reduction in Hb that resulted a decrement in oxygen carrying capacity. The stimulation of the immune system was responsible for enhancement of WBC. The same explanations can also be agreed upon as the explanation for the present study also.
Reported on the fish Labeo rohita due to the exposure difenoconazole (asynthetic pyrethroid) and thiamethoxam (neonicotinoids/new class) of pesticides. The results that are reported in the study are quite contradictory because they had a decrement even in WBC. Even though the reasons were not mentioned in the study however compared to the earlier reports of the study.
In the fish Heteropneustes fossils and Chandna punctate while assessing the toxicity due to the exposure of the lambda cyhalothrin (the group/type I) synthetic pyrethroid similar to the present tested toxicant. The toxicant had several effects including the blood changes in its internal components.
Due to the exposure of the synthetic parathyroid polo (difenthurion–a formulated product of Bifenthrin of the present toxicant) in the fish Labeo rohita (the present tested fish) had an impact on the hematological variations due to the toxic stress. All the parameters decreased including WBC that is different of the present study. The structural damage of the RBC, of hemolysis and hypoxia conditions were the aspects they mentioned as the valid reasons for decrement of RBC and immune response made an impact on WBC. The same might be the same even in the present study.
Due to the pesticide mixture exposure in the fish, Catla catla, Cirrhinus mrigala, Labeo rohita, in which the bifenthrin, (the present studied toxicant) chlorpyrifos (organophosphate) and Endosulfan (organochlorine) had an impact on the enzyme superoxide dismutase in the liver which had a bearing on the metabolism and hematological variations.
Due to deltamethrin in the fish Salmo trutta, in the fish Cirrhinus mrigala in the fish Channa punctatus had the similar results of the present study. But their study pertains only with EC, the commercial formulation whereas the present study pertains to both technical grade as well as 10% EC.
In the fish Salmo trutta fario reported that RBC increased and WBC decreased due to deltamethrin a contrary result of the present study.
In the fish Cyprinus carpio (Common carp) due to cypermethrin toxicity. Considering it as a biomarker study, erythrocytes count, Hemoglobin and the hematocrit values decreased and WBC increased apart from MCHC. Except the MCHC result, the present studied toxicant showed similar results. The stress factor that resulted the health status of the fish disturbed and the very cause of the organism the sustenance which is in jeopardy. Reported the same result in the fish Catla catla due to 10% EC of cypermethrin. The same stress factor was the causative effect that finally led to the death of the fish.
In the fish Labeo rohita (the present studied one) exposed to cypermethrin reported that RBC, Hb and hematocrit, MCH/ MCHC also decreased whereas WBC increased which is similar of the present study of the toxicant bifenthrin. The reasons of the decrement of the erythrocytes include inhibition of synthesis, erythropoiesis and also haem synthesis. The osmoregulatory dysfunction (plasma-formed elements-RBCWBC- platelets) and well as the destruction of the hametopoietic organs are the reasons. The increase of WBC was due to general immune response and defense response and above reasons reiterates the present studied fish for the variations in the blood.
Reported on the bioallethrin another synthetic parathyroid of type I exposed to the fish Channa punctata that due to sub lethal concentrations of the toxic stress the fish blood erythrocytes got damaged in their DNA (Fish erythrocytes are nucleated not enucleated as of mammals) as a result the erythrocytes count decreased.
In the fish Catla catla exposed to cypermethrin (10% and 30% of LC50 values of 0.22 ppm) had an impact on haematological changes. The erythrocytes were damaged by the toxicant which in their view secondary but primarily the tissues of erythropoietin nature was damaged as a result decrement of blood components. The same may be true even in the present study.
Reported in the fish Channa marulus exposed to ¼ of the LC50 value (not 1/10th of LC50 value) of sub lethal concentration the results as of the present study are reported except MCHC.
In the fish Cyprinus carpio, due to cypermethrin toxicity the similar results of the present study are reported.
The effect of deltamethrin in the blood of the fish Cirrhinus mrigala exposing both in lethal and sublethal concentrations and the same results as of above except MCHC of the present study. For the decrease of the RBC, erythrolysis and erythrocytopania were the main reasons of the for it which might be the same even in the present study.
In their review article of the synthetic pyrethroids mentioned some of the different studies and reasons of the alterations were the same which later was mentioned by Sana Ullah, et al., Saha, et al. and Velisek et al.
When the fish Channa punctatus exposed to synthetic pyrethroids plant origin (RUTIN, TARAXEROL APIGENIN and FURADON (Carbamate) that resulted hematological changes similar to the present study. They opined that the stressors (toxicants) evoke non-specific responses in fish and as a result to cope with all the prevailing disturbances to maintain homeostatic state the changes occurred as effects. The same is true even in the present study.
According to due to cypermethrin and deltamethrin where in the study they reported that later was more toxic than former. Leukocytosis and leucopenia were the resultant process and erythropoiesis were the process that is involved in the toxic stress.
Reported in the fish Channa punctatus due to deltamethrin toxicity wherein they studied only two parameters RBC and WBC. The fish reacted in quick time of the stress in order to eliminate the toxicant which in detoxification that had an impact on the blood. Erythropoiesis was inhibited and to have a defense mechanism of homeostasis an aspect of it the WBC increased.
In the same above fish due to the same toxicant of the present studied one reported that the same result of the present study. Hematopoiesis was the reason they explained for the reduction of the RBC and leucocytothermia as for increase of WBC and the same may be true even in the present study.
The poikilothermic blood of the fish if it is altered due to toxic stress had a bearing on the survival even though the concentrations are not lethal. In such sub lethal concentration acting as slow poison, making the fish to suffer as of hematological changes if so, RBC when decreased, oxygen carrying capacity is reduced, metabolism is impaired and growth will be curtailed. WBC changes of increase definitely are immunological response. Both erythrocytopenia and leukocytosis are dangerous for the organism’s wellbeing and that is not good for the species to be cultured. The commercial formulations too significantly altered the blood parameters which have to be noted.
Citation: Rao GS, Rao GS, Rao TS, Rao NG (2022) Haematological Alterations Induced By Bifenthrin (A Synthetic Pyrethroid) Technical Grade And 10% EC in the Fresh Water Fish Labeo Rohita (Hamilton, 1822). J Hematol Thrombo Dis. S3:001.
Received: 02-May-2022, Manuscript No. JHTD-22-17229; Editor assigned: 05-May-2022, Pre QC No. JHTD-22-17229(PQ); Reviewed: 19-May-2022, QC No. JHTD-22-17229; Revised: 26-May-2022, Manuscript No. JHTD-22-17229(R); Published: 02-Jun-2022 , DOI: 10.35248/2329-8790.22.10.001
Copyright: © 2022 Rao GS, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.