**4.3 Mixing of blood of animals and birds with ZnO solution**

From recent studies it is found that most nanoparticles (NPs) show an adverse or toxic effect on blood cells. Studies show that administration of ZnO nanoparticles to whole blood samples and blood serums of animal and bird cause damage to the blood cells and tissues [7–9]. Researchers have investigated that microscopic ultrastructural changes occur in mice when ZnO nanoparticles is incorporated in its body [10].

Zinc oxide solution so prepared is best for use upto one month from the date of preparation but can be used for more number of days if it is kept in colder

**Figure 5.** *Left side: Blood sample of cattle: right side: ZnO mixed with blood sample.*

### **Figure 6.**

*Left side: Blood serum of cattle: right side: ZnO mixed with blood serum.*

temperature. As the solution so prepared is highly acidic (pH = 4.6), so after mixing it with sample of whole blood and serum respectively, it has been observed that colour of the blood and serum changes immediately, it changes from red to pink and colour of serum becomes milky which indicates that haemogolobin of that blood sample decreases as shown in **Figures 5** and **6**.

As pH of blood is 7.2, sodium hydroxide solution is again added drop by drop using micropipette of range 2-20 microlitre to the Zinc oxide solution to make pH value of that solution equal to or greater than the pH value of blood [11]. pH is recorded after every dropwise addition of sodium hydroxide solution to zinc chloride solution which is found as 7.5. After this the solution having pH greater than 7.5 is added to 1ml of blood of cattle. 0.5ml of Zinc oxide solution is added to the same proportion of blood kept for one night. No haemolysis is observed immediately. Again 0.5ml of Zinc oxide solution is added to the same proportion of blood serum of the same cattle and is kept for one night. No haemolysis is observed. But in the next morning again haemolysis is observed in both blood and blood serum.

Due to the occurance of haemolysis, pH of ZnO solution has been made exactly equal to the pH value of blood sample (i.e.7.2). 20μl of NaOH solution is added to 70μl prepared ZnO solution dropwise and after addition it has been observed that pH of ZnO solution becomes 7.2. Again 20μl of the ZnO solution (pH=7.2) is added to1 ml of blood of cattle as well to blood serum and immediately after addition as no haemolysis is observed as shown in **Figures 7** and **8** respectively, so, that sample has been used for manual test of blood to observe the parameters whether any changes have occurred.

And according to the blood test report of bovine given by the Central Instruments Laboratory, College of Veterinary Science, Khanapara, it has been found that RBC(red blood cells) counts shrinks and WBC(white blood cells) are vanished.

Again a new set of experiment is performed with the same ZnO solution. Initially only four parameters of Bovine has been tested. After that, ZnO solution (pH=7.2) of 2μl is added to the 1.5ml of the same blood serum of bovine and is

*Device Diagnosing Health of Bovine DOI: http://dx.doi.org/10.5772/intechopen.99286*

**Figure 7.** *Left side: Blood sample of cattle: Right side: 20μl ZnO mixed with blood sample.*

**Figure 8.** *Left side: Blood serum of cattle: Right side: 20μl ZnO mixed with blood serum.*

**Figure 9.** *Clinical chemistry Analyser (Benesphera).*

tested using Clinical Chemistry Analyzer (Model C-61, Make Benesphera) in the Clinical Laboratory, Khanapara as shown in the **Figure 9**.
