Synthesis, characterization, and use of titanium oxide nanoparticles by pulsed Nd: YAG laser ablation for tick control on domestic animals

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RESEARCH ARTICLE | DE CE MB ER 0 4 20 23
Synthesis, characterization, and use of titanium oxide
nanoparticles by pulsed Nd: YAG laser ablation for tick
control on domestic animals 
Moatasem Al Salih ; Riyad E. Abed; Syakirah Samsudin
AIP Conf. Proc. 2834, 020013 (2023)
https://doi.org/10.1063/5.0161635
05 December 2023 16:00:04

Synthesis, Characterization, and Use of Titanium Oxide
Nanoparticles by Pulsed Nd: YAG Laser Ablation for Tick
Control on Domestic Animals
Moatasem AlSalih1,a( , Riyad E. Abed 2,3,b), Syakirah Samsudin1
1 Biology Dept. Faculty of Science and Mathematics, University Pendidikan Sultan Idris (UPSI), Tanjong Malim
Perak, Malaysia.
2 College of Pharmacy, National University of Science and Technology, Dhi Qar, Iraq.
3 The General Directorate of Education, Dhi Qar Iraq
a) Corresponding author moatasemalsalih@gmail.com
b ) riyad.edanabed@nust.edu.iq
Abstract. This study aims to synthesize acaricide in the form of a new eco-friendly model by using laser technology to prepare a
colloidal solution of titanium dioxide after detecting the presence of a high percentage of resistance to the traditional acaricides
used to treat Ticks and tick-borne diseases. The slope of mortality, lethal concentrations for 50 per centLC50), 90per cent
(LC90), and 9per cent(LC99) with 95per cent confidence interval (CI), and resistance ratios were determined using regression
graphs of probit mortality of larval ticks plotted against log values of increasing concentrations of amitraz (RR50, RR90, and
RR99). The mean RR50, RR90, and RR99 values varied from 0.83 to 1.52, 0.97 to 2.66, and 1.10 to 4.19, respectively,
demonstrating that the field isolates had varying rresistancesTiO2 NPs were created by ablation of a high purity titanium target
(99.99%) submerged in water with a 7ns Nd: YA G laser using atomic force microscopes, the surface morphology of the deposit
materials was investigated (AFM). According to AFM analysis, the average grain size of TiO2NPsinwater was 150 nm, and the
RMS roughness values for TiO2 thin film were (2.27 nm) according to AFM analysis. The solar photovoltaic qualities before and
after Titanium dioxide nanoparticles (NPs) were deposited, as well as the rate of mortalities and time, reveal novel correlations.
our outcomes The potential causes of resistance development are highlighted, as well as recommendations for future brown dog
tick control management programs.
Keywords: Titanium dioxide, Tick Control, Acaricide, Amitraz, lethal concentrations.
INTRODUCTION
Th ree-host tick Hyalomma anatolicum (Acari: Ixodidae) is widely dispersed in Iraq, infecting cattle, buffaloes,
sheep, and goats, as well as parasitic protozoans like Theileria annulata, T. buffet, and T. buffet1]. Tick infection
results in low-quality hides because t of bite marks generated by Hyalomma's longirostrate mouthparts, lower weight
growth,amilk supply productivity and increased calf mortality [2]. Ticks and tick-borne illness management costs
for Iraqi livestock are up to US$498.7 million per year [3-7]. H. anatolicum, the most widespread species in Iraq,
especially Thi-Qar, is reported among 10 Hyalomma species. [8]. An essential instrument for the worldwide control
of ticks were Organophosphates (OP), synthetic pyrethroids (SP), amitraz (Am), and macrocyclic lactones are
examples of chemical acaricides (MLS). However, its widespread and repetitive use has resulted in tick resistance,
toxicity, gastroenteritis, and environmental deterioration [9]. Adaptation to synthetic acaricides in Rhipicephalus
microplus, a single-host animal from Punjab, has been described in several articles, including OP. [10], SP [11], Am
[11], and ivermectin [12]. However, reports of acaricide resistance in H. anatolicum from Iraq are few. [13-15]
Because of the time limit of three-host tick exposure, resistance to acaricide is relatively low in comps compared to
host ticks [11]. In the larval biomass test devised by Stone and Haydock (1962) [16], larvae in a paper packet treated
2nd International Conference of Mathematics, Applied Sciences, Information and Communication Technology
AIP Conf. Proc. 2834, 020013-1–020013-10; https://doi.org/10.1063/5.0161635
Published by AIP Publishing. 978-0-7354-4715-8/$30.00
020013-1
05 December 2023 16:00:04

with technical acaricide were used. The Food and Agriculture Organization (FAO) has now adopted this approach
has the FAO Larval Packet Test to measure tick populations' vulnerability to acaricides (LPT). LPT is being used in
numerous countries to test ticks for OP, SP, and hydrocarbon resistance, but not for amitraz because no amount of
the drug response has been observed. [17]. Accordingly, Miller et al. of the United States Department of
Agriculture, Cattle Fever Tick Research Laboratory (USA) devised a modified LPT for assessing amitraz resistance
in R. microplus, which was then experimentally used by several authors. [18, 19] The investigation was started in
order to calculate the resistance level of amitrate in H. anatolicum ticks in Thi-Qar, Iraq locations using the modified
LPT.
NANOPARTICLES
TiO2 has been researched for a long time. Due to its extensive usage in photo catalysis, optical materials, dye-
sensitive solar cells, and battery sectors [20-24], titanium dioxide is a fascinating family of inorganic solids in a
broad variety of common and high-tech applications. There are three kinds of crystalline TiO2: rutile, anatase, and
brookite, with rutile serving as the thermodynamic stabilizer [25]. liquid (LAL) laser ablation, may lead to
Ethereum conditions and the production of novel nanostructures, pulsed laser removal of metal targets has received
a lot of interest in recent years. [26-30] Composition that is liquid The radiative expansion of the plasma and its
interaction with surrounding media, as well as the locally super high-temperature plasma (approximately 6 000 kb)
and high pressure ((approximately 1 Gpa) the solid-liquid interface, are all produced instantly when a pulsed laser
beam with sufficient energy irradiates into a transparent liquid on a metal target and is quick to tap one pulse about
the same beat. The entire procedure takes 1 second. As a result, laser removal of metal targets in liquid settings
migrespecificationain particular nanomaterials hard to acquire using traditional methods. [31−34].
LAL effectively synthesized rutile TiO2 nanoparticles of a titanium plate in one step at room temperature in distilled
deionized water, according to this report. We performed laser ablation of Titanium Target in water and stored
powder on a solution to generate a highly biological photocatalytic colloidal solution that leads to a concentration
range that is later evaluated for semi-lethal concentration. The surface topology structural characterization of the
deposits was studied using atomic force microscopes (AFM).
EXPERIMENT
Laser ablation of a high-purity titanium target immersed in water at room temperature produced titanium oxide NPs.
A schematic overview of an experimental system configuration is shown in Figure 1. Titanium marks r placed on the
bottom of a 2ml liquid glass sketched was etched laser with wavelength of 1064nm, 7ns, and a repetition frequency
of 1Hz is used to irradiate the Titanium target. To ablate the laser energy of Titanium, a total of 80mJ/pulse was
used for 5 minutes. The laser beam was focused on the titanium target using a 50mm focusing lens. The ablated Ti
NPs were examined using an atomic force microscopy (AFM) model (SPMAA 3000 Angstrom Advanced Inc.
USA).
FIGURE 1. Setup for the experiment B), Colloidal Titanium dioxide NPs freshly produced in water (white-colored).
a
b
020013-2
05 December 2023 16:00:04

MATERIALS AND METHODS
The location, geography, and climate of the research area Thi-Qar Governorate is in southern Iraq, north of Basrah
Governorate, and has internal boundaries with Missan, Muthanna, Qadissiya, and Wassit Governorates. The state of
thi qar is located in southwest Iraq, between latitudes 12.30°N and 12.32°N and longitudes 45. °E. It covers 50,362
square kilometers and is situated between 180 and 300 meters above sea level. The state's average rainfall ranges
from 340 to 512 millimeters, with 120 millimeters in the north and 112 millimeters in the south. Water bodies cover
900 square kilometers, with a length of 120 meters and a width of 110 meters.
Collection Of Ticks
Female ticks that were completely engorged and had fallen were gathered from dairy sheds in al-Nasiriya, al-Rifai,
al-Suq, al-Chibayish, and Al-Shatra in Thi-Qar state's southern regions. Despite the experimental recurring use of
amitraz for tick management throughout the year, tick infestation was identified as a serious concern in dairy cows
in several areas. In cattle shelters, every conceivable tick hiding spot was extensively investigated (cracks and
crevices, loose bricks, and other detritus on the ground). Ticks were collected in plastic vials and covered with a
muslin cloth to enable air and moisture exchange before being transferred to the Entomology Laboratory,
Department of Veterinary Parasitology, Veterinary and Animal Sciences Institute, Thi-qar. The
susceptibility/resistance status in field ticks was determined using the colony of acaricides susceptible reference
IVRI-II line of H. anatolicum.
The Ticks' Preparation
Using a microscope, the ticks were identified as Hyalomma anatolicum. Sen and Fletcher's identification key (1962).
The implanted female ticks were washed in water, drained, and ovipated on 5-cm plastic Petri plates in a moisture
cabinet at 28°C and 85.5% RH. Incubation of the eggs took place under the above-mentioned hatching
circumstances after they had been collected, mixed well, and weighed (20 days). A step-by-step guide to the process
is provided here. An experiment with live larvae aged 14 to 21 days was conducted.
Bioassay
The amended LPT was performed in accordance with Miller et alprocedures. .'s (2002). For the production of
various working concentrations of amitraz, To accomplish serial dilution in two parts, amitraz (Taktic® 12.5 percent
EC, MSD Animal Health, India) and olive oil (OO), Sigma–Aldrich, were diluted to a two-part extent (Sigma-
Aldrich, St. Louis, MO, United States). There were three replicates for each dose with a negative was recorded in
controller (diluents only). The 8 bis 9 cm nylon TChE After receiving 0.6 mL of each dilution attenuators, it was
allowed to evaporate for 2 hours The fabric is a Type 2320 from Cerex Advanced Fabrics in Pensacola, Florida.
After drying, the experimental panels were folded in half and clipped together using metal clips to keep them in
place. Deli No. 8532 (Round Clips) of the Deli Group Co., Ltd. in Ninghai County, Zhejiang Province, China. On
the sides, there is a packet was placed. Each packet was filled with around 100 larvae, and a third clip was placed on
the top. The larvae packages were kept in 24 hours at a rate of 28 ± 1oC, and 85 ± 5% RH. The packages were then
opened and the number of living larvae and dead-rate larvae tallied. Larvae, who moved but didn't move their legs,
had been considered dead.
Statistical Analysis
For probit analysis, Polo-Plus was utilized (Lutilisedftware 2004). The Polo-log-probit Plus model produces a
regression line that illustrates the association between the proportion of larvae that die and the acaricide dosage used
The chi-square test was used to analyze the deviation in the proportionality of log-probit extracted features
(Robertson and Preisler 1992 With a 95 percent confidence interval (CI), the toxic concentrations at 50 per
centLC50), 90 percent (LC90), and 95 paper centLC99) were computed. The variance and covariance of each slope
and intercept were used to calculate resistance ratios (RRs).
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05 December 2023 16:00:04

RESULTS AND DISCUSSION
The color of the solution, which indicates the synthesis of Titanium Oxide Colloid Nanoparticles after laser ablation
of the Titanium target is changed from colorless to candid white as seen shown in Figure (1, b). Figure (2) depicts
3D Two-dimensional and three-dimensional using atomic force microscopy known as (AFM). Titanium oxide NP
images burning in 1m scanned water with a scanning range of 1m per 1m. Its average diameter and height of grain is
about 81.04 nm. For TiO2 thin film, and rm ruggedness values are (2,27 nm). AFM images show that particles with
various sizes have diverse morphologies. Fig. (3, a) illustrates that JSc V is a short circuit current density as an
illuminating energy function before and after the deposition of titanium oxide nanoparticles, at low luminous power
levels, we observed a linearity behavior with a power rise. However, with high lighting power, Jsc has a saturated
behavior that explains, after titanium oxide NP swore 40.2 was deposited, the highest (Jsc) was 40.2. (mA cm-2).
FIGURE (2) AFM pictures of TiO2 NPs that have been ablated in water. A) two dimensions, b) three dimensions of atomic force
The Voce open circuit voltage is a lighting power function before and after the sedimentation process for titanium
particlesic less.N Ps, as shown in Fig. (3, b), and the Voc rise was 630 when the lighting capacity was raised (mV).
The photovoltaic performance is displayed.
FIGURE (3), The diagram here shows the current density Jsc in the illumination simulation before and after sedimentation, as
figure B shows the open-circuit voltage Vocas and the photo simulation function after and before the sedimentation process for
titanium oxide particles.
a
b
a
b
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05 December 2023 16:00:04

Now we have obtained the open-circuit voltage (Voc) of 630mV) which is millivolt and also the amount of electric
current in the impeding circuit which is (current density (Jsc) of 40.2 mA/cm2) and the packing factor (FF=0.7) as in
the figure (4. a, b) where the energy can be calculated from the cell before and after the deposition of titanium
particles. It is most likely that the packing factor is high is the excessive shunt impedance and the short circuit
current density may be due to the effect of photons that are saturated in the depths of the widest part of the silicon
Figures (4 a, b) and (4 c, d) show the output power which is the power generated by the cell under simulation
(AM1) versus the resistive voltage based on the loading impedance before, After the deposition of titanium oxide
particles, it was observed that after the deposition of titanium oxide particles, the efficiency of nanoscale titanium
increased by13.8%. (NPs). After titanium dioxide nanoparticles are deposited, the productivity of the nanoparticles
highly improves.
FIGURE (4) Graph of Titanium oxide NPs photo degradations presentation a) shows before sedimentation NP. b) shows after
sedimentation The output power of Titanium oxide NPs c) before and d) after deposition.
All isolates were shown to be concentration dependent on death, but in controller clusters of numerous samples of
tricks isolates showing impartial to diluents fillers, larval mortality ranged from 0.34 percent to 0.86 percent. Table 1
depicts several isolates of H. anatolicum larvae in a slanted format. data, LC5, LC90, and LC99 (95 % CI), RR50, RR90
and RR99 (95 % CI). The average values ranged between 0.83 and 1.52, 0.97 and 2.66 for the amitraz RR5, RR90 and
RR99 correspondingly. The study demonstrates that among the fieldwork isolated strains, the controllable resistance
station is varied. Four field isolates (Al-Nasiriya, Al-Rifai , Al-Suq , al-Chibayish) demonstrated low resistance
levels based on RR90 and RR99 values, whereas the iso-late Ferozpur represents oversensitive to amitraz. The
d
020013-5
05 December 2023 16:00:04

deterioration schemes of tick larvae's death-rate in contradiction of amitraz dose log are displayed in Figure 1. In Al-
Nasiriya and Al-Rifai isolates, lower mortality slope values indicated heterogee Across the field isolates, there were
several populations with higher RR90 and RR99 values (Figure 2). The sensitive reference regression lines and the
five H. anatolicum field populations were found to have no parallelism or equality (p0.05).
TABLE 1. Hyalomma anatolicum environmental strains were shown to be resistant to the enhanced larval packet test with fatal
dosages of 50 percent, 90 percent, and 95 percent, respectively.
Isolate
N
Slope ± SE
Chi-square
DF
H
LC50 (95%
CI)
LC90 (95% CI)
LC99 (95% CI)
RR50 (95%
CI)
RR90 (95%
CI)
RR99 (95%
CI)
Al-
Nasiriya
133
2.091 ± 0.08
211.0
19
11.1
43.77
179.51
567.26
0.88
1.65
2.75
(32.8–56.8)
(126.2–309.7)
(325.0–1438.0)
(0.80–0.97)
(1.42–1.91)
(2.15–3.51)
Al-Rifai
109
3.133 ± 0.13
60.38
19
3.18
41.17
105.60
227.59
0.83
0.97
1.10
(36.4–46.3)
(89.5–130.4)
(176.5–320.6)
(0.76–0.91)
(0.85–1.11)
(0.89–1.36)
Al-Suq
166
2.190 ± 0.11
68.51
19
3.61
75.15
289.18
867.51
1.52
2.66
4.19
(62.1–90.0)
(220.5–424.8)
(560.4–1664.4)
(1.36–1.69)
(2.22–3.18)
(3.12–5.65)
al-
Chibayish
163
3.093 ± 0.17
178.02
19
9.37
51.56
133.83
291.27
1.04
1.23
1.41
(38.5–66.1)
(98.6–231.7)
(181.7–753.4)
(0.95–1.15)
(1.06–1.42)
(1.11–1.79)
Al-Shatra
208
2.599 ± 0.13
165.43
19
8.71
61.68
191.95
484.32
1.25
1.76
2.34
(47.4–78.3)
(139.1–330.1)
(291.5–1224.8)
(1.13–1.37)
(1.51–2.06)
(1.82–3.02)
Susceptibl
e
197
3.748 ± 0.18
169.92
19
8.94
49.4
108.75
206.62
-
-
-
(40.2–58.9)
(87.4–155.5)
(146.7–385.7)
N: - samples number; SD: - standard division; DF:- freedom degree RR:- resistance ratio, H:- In the case of a proper
test, heterogeneity affects the Chi-square quality; LC = lethal dose in parts per million (ppm); Confidence interval
(CI) is an abbreviated for Confidence Interval.
The excessive humidity, ambient temperatures, and the care of sensitive intercrossbred animals in the area make it
difficult to improve milk production in this location all year (1-4).
Therefore, the frequent use of commercial brands for well-known insecticides with natural and chemical effects is
the most common approach in this field, which is the main entrance to re-arranging a successful strategy to combat
harmful insects, including ticks, of its extensive and indiscriminate use. This has led to increased resistance to those
pesticides and acaricides [12-19.] Despite all this, few cases of resistance to ticks have been recorded, as in the
genus H. anatolicum [7-10,35].
Investigation of the effect of acaricide was prepared as early detection of acaricide resistance in the tick parasite,
which was considered crucial in toxoid the subsequent selection of resistant ticks by using the same active species to
reduce the genetic detection that increases the acaricide resistance capacity and antioxidant secretions in pesticide
physiology, adult age test, age test of Larvae, larval pack test, and larval test are four biological tests that are carried
out in the laboratory to detect true acaricide resistance of ticks to pesticides where according to (Stone and Haydock
1962 ) test (LPT ) was the most common. Filter paper envelopes soaked in oil 100 larvae aged 14–21 days were
injected with a laboratory quality acaricide and kept for 24–48 hours to determine fatality. The test (LPT) was
adapted for early detection of the effect of insecticide resistance, especially amitraz, by replacing filter papers with
nylon sheaths as a basic substrate for the test [15]. The results of this study went with the conclusion reached by
Robertson and Preisler (1992) [36] that the heterogeneity of the recorded data along with the values of LC50 and
LC99 standards of the lethal concentration used, which is where the high results indicate the presence several of
individuals who carry the resistant allele. The current study results show that isolates of ticks with high relative risk
will become more homogeneous but will go towards more heterogeneity than before. Moreover, as a result of testing
in response to acaricide exposure, a heterozygous population will develop for the genes that are resistant to detection
and expression. Genetically, As the frequency of resistant genotypes increases, the gradient will decrease and the
line will shift to the right, indicating a positive correlation. So according to Robertson and Preisler (1992), It has
been found which line represents the representative line (see Figure 1). Amitraz resistance has been demonstrated in
many tick species using the (LPT) test. Many studies have been undertaken worldwide to determine the prevalence
020013-6
05 December 2023 16:00:04

of amitraz resistance in R. microplus. [6-12]. In a recent Ugandan research [19], R. decoloratus and R.
appendiculatus were amitraz resistant. Resistance to amitraz based on AIT has been found in R. microplus from
India. ([17-22]. The present experiments establish special pesticides for parasites such as multi-familial ticks and
hosts, whereas wanted amitraz using a concentration of the least expensive models in the test (LPT assay). and H.
anatolicum species were collected from the same ecological region and worked with other insecticides [11,35]
TABLE 2. Demonstrated the fatal dosage of LC50, LC90, and LC95 and survival and confrontation proportions of 0–36 hours'
larval exposures of numerous Hyalomma anatolicum isolate from the isolates assessed by the improved larval clusters assessment
after titanium exposure.
Isolate
concentration
Number of
Subjects
Observed
Responses
Expected
Responses
Residual
Probability
Al-Nasiriya
1.699
136
22
26.152
-4.562
.192
Al-Rifai
2.000
111
45
38.074
7.298
.343
Al-Suq
2.301
123
63
64.489
-1.814
.524
al-
Chibayish
2.477
112
75
70.612
4.713
.630
Al-Shatra
2.602
120
79
84.073
-5.203
.701
The anti-parasite activity of synthesized composite nanoparticles was tested against five different isolates of Ticks
(al-Nasiriya, al-Rifai, al-Suq, al-Chibayish, and Al-Shatra). Two methods were used in the experiment. The first
method was prepared using aqueous solutions, whose concentrations vary according to the different powers of the
laser. We use between 80 and 200 ml per joule. Then it was kneaded with nanoparticles with the effect of laser
pulses, or what is known as laser ablation. The best results came out of this experiment through nano-recruitment,
which showed the highest level and the best effect as an anti-tick for ticks and metabolic activity, as well as the
physiological impact on the different vital activities through exposure to varying concentrations of nanocomposites
that were attended by the first technique, which is aqueous solutions, which leads to the rapid penetration of body
tissues according to known physical laws such as diffusion and osmosis, as well as the effect of the osmotic pressure
of nano-solutions by increasing the concentration and overlapping of dissolved particles, as the images of techniques
that determine the dimensions of these compounds are clarified by evidence and documents. In addition to all of the
above, the effect of aqueous solutions and the amount of solute in them is added to the presence of oxidative stress
and the voltage generated by the energy gap of the electron, which is the basic principle on which the hypothesis of
catalyzing the inert titanium compound into an effective combination for the ability photodegradation the enzymes
and defense compounds of the parasites is based, which is a developed line of defense in these microorganisms. This
principle focuses on removing or reducing the electron orbit between the valence and the conductor bands. were
examined using a brilliant diffusion technique, which demonstrates that the best dosage of Nanopowder is
400µg.mL-1 Those with the most muscular antiparasite effectiveness is improved and improved. Finally, the activity
of composite nanoparticles of 400g.mL-1 concentration doped with nanoparticles is investigated. The rate of
modality, according to the figure, shows 79 from 120 percent 65. Percent with expected responses of 84.073 and the
probability of 0.701 when using 400 m/ml of titanium dioxide exposure for 12 hours.
Chi-Square
pdf
Sig.
LC50
LC75
LC95
RR2
5.148
3
.161a
182.637
498.933
2118.136
0.945
020013-7
05 December 2023 16:00:04

FIGURE 5. illustrates the efficacy of the nano Acaricide based on where the statistical analysis reveals the activity of the
nanocomposite.e
The above figure 5) shows the effectiveness of the Nano pesticide depending on where the statistical evaluation
shows that the activity of the nanocomposite showed an inhibitory activity for tick growth in a precise ratio between
the observer and the expected. The equation showed that the movement of the Nano acaricide increased at a fixed
rate (y = 3.53 + 1.56x*. In addition to the economic importance of the new compound being cheap, easy to use,
costly to prepare, and environmentally friendly. CONCLUSION
Researchers have created, characterized, and tested titanium oxide nanoparticles for application in tick control in
domestic animals (a new type of acaricide). Pulsed Nd: YAG laser ablation was used as an unique approach to
accomplish this. Another way to this problem is: Pulsed YAG laser ablation of titanium dioxide nanoparticles was
employed in this experiment. The antioxidant system was photodegraded and the oxidative responses were estimated
using these nanoparticles. A novel species photocatalytic visible-light activity of co-doped TiO2 with a wide range
of pulses was investigated. Atomic force microscopes were used to examine the deposit materials' surface
morphology (AFM). Using AFM, it was found that the TiO2NPs in water had an average grain size of 150 nm and
an RMS roughness value of 2.27 nm. The AFM was used for both of these measurements. Before and after titanium
dioxide nanoparticles (NPs) were applied to the solar photovoltaic characteristics, the bioassay of tick mortality rates
and time against five separate isolates of ticks indicated unique associations. as a result of This article sheds insight
on some of the factors that may have contributed to the rise of the resistance. To investigate the anti-parasitic
properties of manufactured nanoparticles, Finney used five different tick strains. Log-probit linear regression was
used to test the validity of the assessment (1971). TiO2 as an acaricide was studied using the equation y = 3.53 +
1.56x*, which showed that the growth of the Nano acaricide was constant. Besides being affordable, user-friendly,
environmentally benign, and costly to produce, the new material has economic importance.
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