*3.1.3 Interaction of Chlorpyrifos and Cypermethrin with bovine hemoglobin and bovine serum albumin*

Titration experiments showed that the fluorescence intensity of the BSA gradually decreased while the fluorescence intensity of the reaction system containing BHb3 increased gradually due to interaction with cypermethrin. The maximum emission wavelength was constant at around 340 nm. That is, there was no red or blue shift.

<sup>3</sup> Bovine hemoglobin

Finally, Chlorpyrifos and Cypermethrin were able to bind BSA and Bovin Hb, and both pesticides bind to Albumine much more potent than that hemoglobin [25].

#### *3.1.4 Interaction of Paraquat with bovine hemoglobin*

The reactivity of the heme center with the superoxide anion formed by paraquat is judged by the decrease in the Soret band, and all four heme groups associated with hemoglobin are damaged and eventually destroyed by the superoxide anion formed by the PC.

UV/Vis absorption and synchronous fluorescence spectroscopy revealed that the environmental structure of these Trp (tryptophan) residues was altered and that the results showed that the presence of one class of binding sites on BHb, hydrophobicity, and electrostatic interactions play an essential role in the stabilization of the complex [26].

#### *3.1.5 Interaction of Imidacloprid with hemoglobin*

Ding et al. [27] investigated the binding of Imidacloprid with hemoglobin. They showed that Imidacloprid quenched hemoglobin's intrinsic fluorescence via the static quenching process. The values of enthalpy (ΔH = −14.58 kJ mol−1) and entropy (ΔS = 32.83 J mol−1 K−1) of the reaction indicate that hydrophobic interactions and hydrogen bonding are the dominant intermolecular forces in stabilizing the Imidacloride-Hb complex.

### **4. Insecticide resistance**

There are two mechanisms for insecticide resistance: behavioral and physiological. In behavioral resistance, the insect's reaction reduces or prevents exposure that can lead to death. Otherwise, there are different types of modification mechanisms in physiological resistance like decreasing cuticular penetration and target site sensitivity or increasing metabolic detoxification [28]. To explain more, a common feature of insecticide metabolic resistance is the overexpression of detoxification genes at the transcriptional level, leading to high levels of protein and enzymatic activity. Therefore, this detoxification and resistance development level [29]. One of the notable examples of physiological resistance is malaria, which still exists in some African countries like Tanzania, while chemical insecticides are used against them. The straightforward reason is that target-site insensitivity (knockdown resistance' target-site mutations) in malaria vectors, lower penetration, or an enhanced detoxification activity [30].
