Fenoxycarb, an Insect Growth Regulator (IGR)
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Transactions on Ecology and the Environment vol 8, © 1996 WIT Press, www.witpress.com, ISSN 1743-3541 Fenoxycarb, an insect growth regulator (IGR), induces anemia in mammals A.A. Bazzaz Department of Environmental and Evolutionary Biology, University of Liverpool, Liverpool L69 3BX, UK Abstract Fenoxycarb, an Insect growth regulator (IGR), has been investigated to determine its toxic effect on mammalian life by analyzing blood samples as an indicator. One hundred and eight male mice were used in this study (24 control and 84 experimental animals). The mice were sacrificed by head dislocation, and blood samples were collected into heparinised tubes for haematological tests. Haematological changes in blood parameters have been measured in mice given fenoxycarb in water at three different dose (1, 5 and 10 ppm) for 4 weeks. Fenoxycarb primarily induced a mild to moderate anemia as early as the first week, at all three doses by causing a significant (p<0.05-0.0001) decline in the levels of haemoglobin (Hb%), packed cell volume (PCV%) leucocytes (WBC) and erythrocytes (RBC) population. It also caused a steady decline in both macrophages and neutrophils. However, changes in other leucocyte counts were inconsistent. It has been concluded that fenoxycarb, at the doses used in these experiments, is not directly toxic, but long term exposure could lead to hypoxia stress and an increased risk of infection. Introduction It has been found that insect growth regulators (IGRs) are less dangerous than pesticides to non-target animals and to humansi. The hazards of the pesticides and IGRs to aquatic organisms have well been documented2,3. Recently, a new compound with juvenile-hormone [JH] activity, ethyl (2-(4-phenoxy-phenoxy) ethyl} carbamate [fenoxycarb], was reported to exhibit a strong juvenile-hormone activity against a variety of insects^-io. it is a non-terpenoid, non-neurotoxic carbamate and has been shown to interact with juvenile hormone esterase activityn. The activity of IGRs is reported to be via different modes of Transactions on Ecology and the Environment vol 8, © 1996 WIT Press, www.witpress.com, ISSN 1743-3541 374 Air Pollution Monitoring, Simulation and Control action, and various methods have been employed to control the growth of insects and arthropods. Moreover, the IGRs have greater field stability and, against some insects, higher potency and control potential than the earlier terpenoidis.ia. However, when these IGRs run off to adjoining aquatic ecosystem, they cause some hazards to aquatic animals as well as to non-target animals by accidental contact with surface water of industrial and agricultural areas^. One of the principal advantages of using IGRs instead of pesticides in insect control is their very low mammalian toxicityio. They are not necessarily immediately toxic but eventually cause abnormalities that impair survival or reproduction of insects and arthropods! 5-17. To the best of my knowledge, nothing is known about the effect of fenoxycarb on the non-target animals which might drink contaminated shallow ground water. The present study has been designed to investigate the haematological effects of fenoxycarb to laboratory animals (mice) as an example of non-target mammals. Materials and Methods Preparation of Fenoxycarb Doses: One hundred and eight male albino Swiss mice (Mus musculus) weighing (25±2 gm) were used in this investigation. Their age ranged from 3 weeks old at the beginning of the experiment up to 2 months old at the end of week four. They were maintained under controlled environmental conditions of 25±2°C. Woodshavings were used as bedding material. Normal laboratory chow and tap water (except for treated animals) were available ad lib and were replenished three times a week. Fenoxycarb was obtained as a gift from Maag Agrochemicals as a pure solution, and was stored at 4 C. Three concentrations: 1, 5 and 10 ppm were emulsified with tap water and were prepared freshly three times a week and used as replacement for drinking water for the experimental animals. Haematological Tests: At the end of each week, twenty one experimental animals (7 per group) in addition to six (control mice) were scarified by cervical dislocation (without anesthesia). Blood samples were collected from the cervical artery into heparinised tubes for haematological tests. Levels of haemoglobin in (g/dl), packed cell volume (PCV%, haematocrit) and differential counting of leucocytes (absolute numbers was performed according to method described by BatraiG. Counting of leucocytes and erythrocytes was done using haemocytometer. Blood smears were prepared on slides and stained with Lieshman's stain. The blood cell counting performed using Olympus light microscope (100x objective oil lens). Biostatistical analysis were carried out using student T-test. Transactions on Ecology and the Environment vol 8, © 1996 WIT Press, www.witpress.com, ISSN 1743-3541 Air Pollution Monitoring, Simulation and Control 375 J2 <D I Control/W1 Experl./W1 Cont./W2 Experi./W2 Control/W3 Experl/W3 CO Control/W4 0) Experl./W4 O) 0) 2 0) Q. Hb% (1ppm) Hb% (Sppm) Hb% (10ppm) Figure 1: Dose and time-dependent decrease in Hb%. 50 Control/W1 40 - Experl./W1 Cont./W2 - 30 H Experi./W2 o a. Control/W3 •5 20- Experl/W3 0) o> «#COw Control/W4 10 - Q) Exper!./W4 2 <D Q. PCV% (1ppm) PCV% (Sppm) PCV% (10ppm) Figure 2: Dose & time-dependent decrease in PCV%. Transactions on Ecology and the Environment vol 8, © 1996 WIT Press, www.witpress.com, ISSN 1743-3541 376 Air Pollution Monitoring, Simulation and Control 0) o> • Control/W1 C3 Exper!./W1 • Cont./W2 0 Experi./W2 o 10- o H Control/W3 Q Experl/W3 H Control/W4 H Experl./W4 0 CO CD 0 * WBC% (1p|WBC% (5pWBC% (10ppm) Figure 3: Decrease in WBC mean numbers of treated vs control animals • ControlAVI Q Experl.W1 H Cont./W2 El Experi./W2 • Control/W3 Ei Experl/W3 • Control/W4 H Experl./W4 RBC (1ppm)RBC (SppmRBC (10ppm) Figure 4: Dose and time dependent decrease in number of RBC. Transactions on Ecology and the Environment vol 8, © 1996 WIT Press, www.witpress.com, ISSN 1743-3541 Air Pollution Monitoring, Simulation and Control 377 Tablel: Blood parameters of Mice given 1 ppm fenoxycarb in the drinking water. (Hb%) percentage of Haemoglobin; (PCV%) percentage of Packed Cell Volume; (WBC) number of leucocytes; (RBC) number of erythrocyte; (SD) standard deviation; (p) Student T-test. Animals Mean Hb% Mean PCV% Mean WBC & Mean RBC & Weeks(W1-W4) ISO ISO & ±SD & ±SD (x1 03/u/) (x1 QG/u/) Control (n=6) 13.55±0.696 44.50±2.22 9.0510.333 9.5010.327 Treated (n=7) 13.38+0.042 42.00±2.097 7.8010.335 9.6410.320 P< NS NS 0.005 NS Control (n=6) 13.7010.541 43.80±3.16 9.1010.333 9.7010.34 Treated (n=7) 11.18±1.772 37.35±0.599 7.8110.362 9.6410.373 0.05 0.005 0.0005 0.005 P< Control (n=6) 11.20±0.42 39.2011.24 10.012.72 9.8910.343 Treated (n=7) 9.85±0.407 34.6310.953 8.1210.137 8.2010.907 FK 0.05 0.005 0.005 0.013 Control (n=6) 13.82±1.12 44.6710.55 8.3013.82 9.3110.17 Treated (n=7) 9.6±0.314 33.9111.297 7.5211.34 7.2210.323 0.0005 0.005 0.005 P< 0.005 Table 2: Blood parameters of Mice given 5 ppm fenoxycarb in the drinking water. (Hb%) percentage of Haemoglobin; (PCV%) percentage of Packed Cell Volume; (WBC) number of leucocytes; (RBC) number of erythrocyte; (SD) standard deviation; (D) Student T-test. Animals Mean Hb% Mean PCV% Mean WBC & Mean RBC & Weeks(W1-W4) ISO 1SD &1SD & ISO (x1 03/t//) (X106/CI/) Control (n=6) 13.5216.96 44.5012.22 9.0510.333 9.5010.327 Treated (n=7) 12.7010.357 40.8011.326 7.5010.368 8.6610.338 P< NS 0.01 0.0005 0.004 Control (n=6) 13.7310.541 43.8013.16 9.1010.333 9.7010.34 Treated (n=7) 11.1510.55 36.3511.32 7.7510.83 8.2810.31 0.05 0.05 0.0005 0.005 P< Control (n=6) 11.2010.42 39.2011.24 10.012.72 9.8910.343 Treated (n=7) 9.6010.365 36.9311.289 7.4710.37 7.5210.157 0.005 P< 0.05 0.005 0.005 Control (n=6) 13.8211.12 44.6710.55 8.3013.82 9.3310.17 Treated (n=7) 9.4010.19 32.0210.821 7.8710.188 6.4210.219 0.005 0.005 0.005 0.0005 P< Transactions on Ecology and the Environment vol 8, © 1996 WIT Press, www.witpress.com, ISSN 1743-3541 378 Air Pollution Monitoring, Simulation and Control Table 3: Blood parameters of Mice given 10 ppm fenoxycarb in the drinking water. (Hb%) percentage of Haemoglobin; (PCV%) percentage of Packed Cell Volume; (WBC) number of leucocytes; (RBC) number of erythrocyte; (SD) standard deviation; (p) Student T-test. Animals Mean Hb% Mean PCV% Mean WBC & Mean RBC & Weeks(W1-W4) ISO ISO &±SD & ±SD (X1Q3/W/) (X106/U/) Control (n=6) 13.52±6.96 44.50±2.22 9.0510.333 9.5010.327 Treated (n=7) 11.30±0.681 36.67±3.15 7.1710.17 8.1810.14 0.0005 0.001 0.0005 0.0005 P< Control (n=6) 13.73±0.541 43.80±3.16 9.1010.333 9.7010.34 Treated (n=7) 9.38±0.552 32.78+1.297 7.3010.227 7.3010.33 P< 0.0005 0.0005 0.005 0.005 Control (n=6) 11.20±0.42 39.2011.24 10.012.72 9.8910.343 Treated (n=7) 9.50±0.224 32.65±1.732 7.3010.153 7.0810.397 0.05 0.05 0.005 0.05 P< Control (n=6) 13.82±1.12 44.67±0.55 8.3313.82 9.3310.17 Treated (n=7) 9.40±0.224 31.4011.36 7.7210.414 6.2710.345 P< 0.0005 0.005 0.005 0.0005 Table 4: Differential leucocyte counting (100u/) of Mice given 1 ppm fenoxycarb in the drinking water.