Toxicological Studies on Boric Acid, Imidacloprid And
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TOXICOLOGICAL STUDIES ON BORIC ACID, IMIDACLOPRID AND FIPRONIL AND THEIR BINARY MIXTURES AS INSECTICIDES ON GERMAN COCKROACH Blattellagermanica (L.) (DICTYOPTERA: BLATTELLIDAE) By FATMA SHERIF AHMED B.Sc. Agric. Sci. (Pesticides), Fac. Agric., Cairo Univ., 2007 THESIS Submitted in Partial Fulfillment of the Requirements for the Degree of MASTER OF SCIENCE In Agricultural Sciences (Pesticides) Department of Economic Entomology and Pesticides Faculty of Agriculture Cairo University EGYPT 2015 ١ INTRODUCTION The German cockroaches, Blattellagermanica (L.), (Dictyoptera, Blattellidae) are the most common indoor pests, especially in multiple-family housing and the most significant pest in many parts of the world (Goddard, 2003). German cockroaches prefer warm, wet locations with high humidity such as kitchens, bathrooms and laundry areas. These conditions are available in several places as homes, apartments, restaurants, supermarkets, hospitals and other buildings where food are stored. Cockroaches are not only corrupt food but also transfer pathogens such as Salmonella, Shigella, Escherichia coli, Staphylococcus aureus and Bacillus cereus (Baumholtz et al., 1997 and Tachbeleet al., 2006). Medically important parasites such as bacteria, fungi and molds, protozoans, viruses were isolated from external and internal surface of cockroach (Brenner, 1995). Cockroaches can also transfer both gram-positive and negative bacteria (El-Sherbini and El- Sherbini, 2011). A large number of neurotoxic and non-neurotoxic insecticides were used for German cockroach control, as this pest has a considerable ability to develop resistance to a variety of chemical insecticides(Cochran, 1989 and 1995a; Scott et al., 1990; Rust and Reierson, 1991; Rust et al., 1993; Holbrook et al., 1999; Espinosa-Islas et al., 2002 and Rahayuet al., 2012). The toxicity of some new neurotoxic insecticides is either very limited (i.e., neonicotinoids) or not well established (i.e.,phenylpyrazole) against German cockroach in Egypt. Boric acid (H3BO3) has been used as an insecticide since 1948 (See et al., 2010). It was the standard chemical used for the control of some pests until the chlorinated hydrocarbon pesticides become available (Lal and Srivastava, 1950). Moreover, it is not classified as carcinogen or skin irritant (Langford et al., 1954). One advantage of using boric acid is that, it destroys the cellular lining of the foregut. This effect induces death to the insects by starvation and ٢ the cockroaches have difficult to show any resistance to boric acid (Cochran, 1995b). A phenylpyrazole insecticide fipronil discovered by Bayer Crop Science in 1987, and introduced in 1993 as a form of solid (e.g., insect bait), liquid spray, or as a granular product (Anonymous, 2000), and registered in the U.S. in 1996 (Ware, 2000 and Jeong Lee et al., 2010). It controls cockroaches, mosquitoes, ticks, and fleas at both larval and adult stages (Chantonet al., 2002). Cole et al. (1993) found that fipronil interferes with the δ-aminobutyric acid (GABA) channel, where it disrupts normal nerve influx transmission (e.g., passage of chloride ions) this causes excessive neural excitation, paralysis, and finally insect death (Gant et al., 1998). Fipronil gel baits have been widely used to control German cockroach in America since several years due to german cockroach resistance to several insecticides and fipronil gel bait consider more safe in application than other treatment methods (Ahmed and Suliyat, 2011). A neonicotinoid insecticide, imidacloprid belonging to the chloronicotinyl compounds. It was first proposed in 1991. It exerts toxic effects after oral ingestion; it is also suitable for use in bait formulations (Nasirian, 2007). The main factor of chloronicotinyl being favorable is their low affinity to vertebrate compared to insect nicotinic receptors. They have high selectivity; have a quick knock down effect and long residual activity. Worldwide, it is considered to be one of the insecticides used in the largest volume (Cox, 2001). Nasirian (2008) reported that fipronil and imidacloprid gel baits completely killed the German cockroaches under laboratory conditions in ingested bait method. Poisoned baits, in the last two decades, have largely displaced other formulations for controlling of German cockroaches and other pests (Anaclerio and Molinari, 2012). The poisoned baits are much less translocatable, so it is less hazardous than the spray formulations (Gore and Schal, 2004). So that the tested insecticides used as a bait to be safer and environmentally friendly than ٣ insecticide sprays. The toxicity of fipronil baits has been estimated from mortality rates (Scott, 1991 and Ross, 1993) and consumption rate (Wang et al., 2006). The three tested compounds (boric acid, imidacloprid and fipronil) were selected on the basis of their high efficacy against German cockroach. Although fipronil hasn't been used in Egypt against German cockroach yet, it widely used in all developed countries as a highly effective tool for controlling the German cockroach especially in bait form. The present study evaluated the efficacy of fipronil, imidacloprid and boric acid on the nymphal instars and adult males of German cockroach. On the other hand, German cockroach didn't appear to be resistant to boric acid compound although this compound is used against this insect for ages. Also, the application of neonicotinoid insecticide imidacloprid in bait form is easy to apply in out-of- way places and is stable under a wide range of conditions. The objectives of current study are: 1. Determination of the oral toxicity of boric acid, imidacloprid and fipronil as poisoned baits against different stages of B. germanica (L.). 2. Evaluation of the palatability and the toxicity of fipronil incorporated into different food baits on the 6th instar nymphs of German cockroach B. germanica (L.). 3. Comparing the toxicity of imidacloprid as topical application and stomach poisoned bait against the different stages of the tested insect. 4. Estimation of the lethal times (LT50) of the two tested compounds on the different stages. 5. Evaluation of the effect of binary mixtures of the three tested insecticides on 6th instar nymphs. ٤ MATERIALS AND METHODS Tested insect (Rearing and Manipulation the Insect): German cockroach, BlattellaGermanica (L.) Family: Blattellidae Order: Dictyoptera Cockroaches are insects with three stages: eggs, nymphs, and adults, males and females. About one week after mating, the female cockroach deposits eggs into a protective covering called the oetheca or egg case which protects the eggs from drying. The egg case looks like a tiny, brown, purse-shaped capsule, it is about 8 mm long, 3 mm high, and 2 mm wide (Tanaka, 1976). The eggs hatch into a small but fully-developed nymphs that grow through a series of molts, shedding its old cuticle (exoskeleton) at each molt. The range of head capsule widths in the lst to 6th instars nymphs are, respectively: 0.74-0.85 mm; 0.88-1.05 mm; 1.02-1.35 mm; 1.20-1.62 mm; 1.40-1.86 mm; and 1.84-2.25 mm (Murray, 1967). Nymphal survival percentages from egg hatch to the adult stage was estimated at 85- 87% (Willis et al., 1958 and Archboldet al., 1987). The nymphs of cockroaches emerge as fully-winged adults after the final sixth molt. Newly emerged adults appear white, but become fully pigmented within 24 hours (Ross and Mullins, 1995). As the nymphs develop faster, they gain more body mass and the adults are heavier, the females produce more fertile oethecae (Gemenoet al., 2001). German cockroaches are semi-social insects; all their stages and ages are found at the same place. Colony of German cockroach BlattellaGermanica (L.) used for the present study was obtained from Aburwash, Giza Governorate, reared for two years and the all bioassay tests were carried out in the Department of Economic Entomology and Pesticides, Faculty of Agriculture, Cairo University, Egypt. The average number of (eggs per capsule, capsules per female), period between egg case formation to hatching, number of molts, ٥ nymphal stage (in days), life cycle and average longevity of adults (in days) are given in the following. Cockroaches insects were reared in plastic containers (1 liter capacity) and maintained at 27 ± 1°C, 60±5% relative humidity in an incubator at a photoperiod of 12:12 dark-light. A wet piece of cotton was placed in each jar as a water source for the cockroaches with some of modifications as described by (Nalyanya and Schal, 2001; Nasirianet al., 2006a and 2011; Sulaimanet al., 2007 and Kilani-Morakchiet al., 2009). The cockroaches for rearing were fed on a source of protein and a sugar solution coated on a piece of cotton, and a cardboard or a plastic tubes as a shelter (Nasirianet al., 2006a,c). The present investigations were carried out on the 2nd, 6th instar nymphs (two days old) and adult males (2-3weeks old). Only adult males, not adult females, were used in the experiments because their weight and physiology are more uniform than adult female (Appelet al., 1983). The females were also needed for further reproduction. Insecticides tested: Three commercial formulations were used in the toxicological study. These compounds are representing boric acid (inorganic compound), imidacloprid(neonicotinoid insecticide group) and fipronil (phenylpyrazole group). The common name, trade name and properties of the tested compounds (boric acid, imidacloprid and fipronil) are given in Table 2. a. Boric acid insecticide: Boric acid (H3BO3) was used as an insecticide for control of cockroaches, termites, fire ants, fleas, silverfish, and many other insects(See et al., 2010). It is considered to be safe to use in household kitchens to control cockroaches and ants.The dry powder is abrasive to the insect and may also have some toxic effects on the nervous system of insects (Habeset al., 2005). It acts as a stomach poison (affecting the insect’s metabolism) and as a contact poison. ٦ b. Imidacloprid insecticide: Imidacloprid is an insecticide belonging to the neonicotinoid class of compounds. It was first proposed in 1991 and registered for use by the United States Environmental Protection Agency (U.S.