Anti-cockroach Activities of Biosynthesized Silver Nanoparticles using Petiveria alliacea Extracts

doi:10.21203/rs.3.rs-5253110/v1

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Anti-cockroach Activities of Biosynthesized Silver Nanoparticles using Petiveria alliacea Extracts

https://doi.org/10.21203/rs.3.rs-5253110/v1

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Background

This study was conducted with a view to biosynthesize silver nanoparticle (AgNPs) using the root (PaR) and leaf (PaL) extracts of Petiveria alliacea and evaluation of their activities against cockroach which is a major household insect pest.

Methods

The biosynthesized AgNPs were characterized by UV-Vis spectrophotometry, Fourier transform infrared (FTIR) spectroscopy and transmission electron microscopy (TEM). PaR-AgNPs and PaL-AgNPs were tested at three different concentrations (1, 5, 10% v/v) using two mode of applications (fumigant and contact toxicity) at different hours of exposure.

Results

The peak absorbance of the AgNPs occurred at the wavelengths of 426 and 442 nm for PaL-AgNPs and PaR-AgNPs respectively with the brownish colloidal solutions. FITR peaks indicated the presence of alkenes (= C-H-), esters (C = O), amides (N-H), and aliphatic amines (C-N) in the samples, while the TEM and EDX analyses showed the presence of silver in the AgNPs colloidal solution with the spherical nanoparticles ranging from 5.95–76.19 nm in size. Both AgNPs exhibited insecticidal action against adult cockroach, with the fumigant application exerting killing faster than the contact application. While 80–90% mortality of cockroach was observed through fumigant, the contact application resulted to 40–50% mortality after three hours of exposure at 10% v/v. These show that the effectiveness of the bio-fabricated AgNPs formulations depended on the concentrations, hours of exposure, and mode of application.

Conclusion

Therefore, silver nanoparticles based insecticides can be incorporated into the pest management program of household pests especially cockroach.

Cockroach

Petiveria alliacea

Silver nanoparticles

Biosynthesis

Nano-insecticide

Fumigant

Contact insecticide

• TEM and EDX analyses indicate the presence of silver in the nano-insecticide products

• Biosynthesized AgNPs exhibited insecticidal action against adult cockroach

• Toxicity effect of nano-insecticides depends on mode of application

• Nano-insecticides applied through fumigant killed faster than when applied through contact

• Effectiveness of biosynthesized AgNPs was dose-dependent

Cockroach is one of the major household insects throughout the world [1]. Specifically, Periplaneta americana (L.) is one of the six dominant cockroaches in Nigeria, especially South Western part. It is commonly found in residential buildings most especially in the kitchen and toilet, as well as in the restaurants, hotels and bakery [2]. Infestation of cockroaches has health implications in the transmission of diseases by contact with human foods [2, 3]. P. americana is a coprophagous vector, contaminating large quantity of consumable by leaving behind human faecal matter and parasitic cyst/eggs of helminths and protozoa [2]. According to Ruhma et al. [4], cockroach infestations can result in allergic reaction in some people and such reaction may include asthma and skin irritation [5].

In view of the aforementioned problems, the use of synthetic insecticides has been the most common strategy to curtail the infestation because of their quick action against cockroaches. The major classes of synthetic insecticides are organochlorine, organophosphate, carbamates and pyrethroid [6, 7]. These classes of insecticides are relatively stable in the environment thereby constituting environmental and health hazards [8, 9]. Existing studies have shown that some species of cockroaches have developed resistant to synthetic insecticides [10, 11]. Also, availability of these insecticides at a critical need has become a serious concern, coupled with their increasing costs [12, 13]. All these associated problems with synthetic insecticides bring to the limelight the idea of alternatives to synthetic insecticides; notably biopesticides and nanopesticides.

Biopesticides are defined as biological pesticides which can be made from plants, microorganisms and animals [14, 15]. They are also described as natural chemicals from the natural sources which are used as alternatives to synthetic chemicals in the control of pests [16, 17]. Biopesticides are environmental-friendly, can be easily produced with low cost, and are effective at killing target pests [18–20]. Nano-pesticide is categorized as biopesticides due to its characteristic nature with Biopesticides [14]. Nanoparticles in the formulations of Nano-insecticides can act as adjuvants thereby improve the efficacy of insecticides (Manna et al. 2023). Nano-insecticides formulation can be a mixed formulation of metal and other various materials that have been established against a given pest [21, 22].

P. alliacea L. belongs to the family of Phytolaccaceae which are found abundantly around South and Central America tropics, Africa and Caribbean [23]. Several chemical compounds have been isolated from P. alliacea which include: astilbin, myricitrin, engeletin (flavonoids), triterpenes, daucosterol and lignoceric acid [23]. Meanwhile, dibenzyltrisuilphoid and dichloromethane are part of the chemical compounds which have pharmacological and insecticidal effects [23–26]. However, there is dearth of information on the production and evaluation of nano-insecticides from P. alliacea and this is the first reported case of Pa-AgNPs in the control of cockroach.

2.1 Materials

Leaf and root extracts of P. alliacea, silver nitrate, filter paper, insect box, syringe, cotton wool, buckets, transparent containers, paper tape, biscuits, pineapple and insect rearing wooden cage.

2.2 Test Insect Collection

The cockroaches (P. americana) were collected from slums, dumping grounds and several households in Ogbomoso city in Oyo State, South-western Nigeria, between April, 2021 and July, 2021. They were introduced into insect wooden cage boxes (20 cm x 15 cm x 5 cm) and rags of different sizes were placed into each box to help mimic the natural environment of the cockroach. Feed consisted of biscuits and pineapple juices were evenly mixed together and these were given as feed. Each wooden cages consisted 25 adult unsexed cockroaches and they were allowed to acclimatize to their new environment for a month. Healthy cockroaches were picked from the insect box irrespective of the gender and used for the study.

2.3 Biosynthesis and Characterization of Silver Nanoparticles

Leaves and roots of P. alliacea were harvested and air-dried under laboratory conditions for 4 several weeks. The dried plant parts were blended separately using electronic grinding machine (Silver Crest 50 watts) into fine particles.

Silver nanoparticles: Nano P. alliacea leaf (PaL-AgNPs) and Nano P. alliacea root (PaR-AgNPs) used as nano-insecticides in this study were synthesized using both leaf and root extracts of P. alliacea following the modified methods of Lateef et al. [27]. Extracts were prepared by heating 5 g of the powdered sample (leaf or root of P. alliacea) dissolved in 100 ml of distilled water at 60 ^oC for 1 h, then filtered through Whatman No.1 filter paper and the filtrate was subsequently centrifuged at 4000 rpm for 30 min to obtain the respective extract. The biosynthesis were then carried out by reacting about 2 ml of aqueous extract with 40 ml of 1 mM silver nitrate (AgNO₃) solution for the synthesis of AgNPs at ambient temperature (30 ± 2 ^oC) under bright light condition. The biosynthesis was monitored by observing the change of colour of the solution.

Moreover, characterization of the biosynthesized AgNPs was carried out by measuring the absorbance spectrum using UV–Vis spectrophotometer. Fourier transform infrared (FTIR) spectroscopy analysis was done to establish the biomolecules responsible for the synthesis, capping, and stabilization of AgNPs. As described by Lateef et al. (2017), the AgNPs were further observed on the transmission electron microscope (TEM 1400, JEOL, USA) to determine the size and morphology couple with the selected area electron diffraction (SAED).\

2.4 Bioassay

2.4.1 Contact toxicity bioassay

This experiment consisted of eight units made up of four healthy adult unsexed cockroaches picked from the insect wooden cage box. It commenced in the morning at 8 am. From the stock solution, 1, 5 and 10% v/v were prepared by diluting the equivalent volume of the stock solution with 100 ml of water and 5 ml of each tested concentration was applied to the studied insect. A calibrated syringe was used to measure 5 ml of the prepared concentrations (1, 5 and 10% v/v) of PaL-AgNPs and PaR-AgNPs formulations needed for the study. Each experimental unit was well labeled with the corresponding treatment names viz PaL-AgNPs, PaR-AgNPs, synthetic insecticide (lambdacyhalothrin) and control (water). Diluted lambdacyhalothrin (1 ml of lambdacyhalothrin into 2000 ml of water) was applied as positive control. The solution drawn with the medical syringe (5 ml) was sprayed directly on the cockroaches in each designated experimental unit. Each treatment was replicated thrice and mortality was recorded at hourly interval for 20 h. Each of the treatments was replicated thrice.

2.4.2 Fumigant Toxicity bioassay

The transparent containers (8 units) used for each experimental unit were weighed with a sensitive scale and 0.1 g of cotton wool was placed into each of the six containers used for this experiment. Then, 5 ml of the tested concentrations (1, 5 and 10% v/v) of Nano P. alliacea solution was applied to the cotton wool (8) in each designated container separately. Water was applied as negative control to the studied insect, while diluted lambdacyhalothrin as previously stated was applied as positive control. The four active adult unsexed cockroaches were then introduced into each assigned containers and mortality was recorded every one hour for 20 h. The experiment was replicated thrice for each treatment.

2.5 Data collection and statistical analysis

The mortality of the studied insect was determined and later subjected to percentage mortality using Abbot’s formular (1925):

%Mortality = \(\:\frac{CA-TA}{CA}\) × 100

Where, TA is the number of dead treated cockroach; CA is the number of introduced cockroach

While the percentage mortality was corrected using Abbot’s formular (1925):

Pr = \(\:\frac{Pa-Pc}{100-Pc}\) × 100

Where, Pr is the percentage of efficacy of the insecticide, Pa is the percentage mortality of the treated insect, and Pc is the percentage mortality of the untreated insect.

Percentage mortalities were later subjected to analysis of variance (ANOVA) using statistical software analysis (2005) version 2.0. Means were separated using least significant difference (LSD) at 5% probability.

3.1 Biosynthesis and Characterization of AgNPs

The leaf and root extract of P. alliacea biosynthesized AgNPs (Nano P. alliacea leaf (PaL-AgNPs) and Nano P. alliacea root (PaR-AgNPs) in 10 min under ambient conditions of temperature (30 ± 2°C) with typical dark brown colour for PaL-AgNPs and light brown colour for PaR-AgNPs (Fig. 1) which deepened over time and stabilized eventually after 25 min. The brownish colored biosynthesized AgNPs have been extensively reported [27–29]. Moreover, maximum absorbance occurred at the wavelengths of 426.32 and 442.11 nm for PaL-AgNPs and PaR-AgNPs respectively (Fig. 2). Absorbance at these wavelengths is due to the unique surface plasmon resonance of the biosynthesized AgNPs and the values here reported are within the range of 391–460 nm previously established in related existing literatures for AgNPs [28, 30].

The FTIR absorption spectra revealed distinct and prominent bands at 3030, 2121, 2013, 1938, 1745, 1575, 1284, and 643 cm^− 1 for PaL-AgNPs, which are observed to be close to 3029, 2108, 1933, 1576, and 1222 cm^− 1 obtained for PaR-AgNPs (Fig. 3). Also, other closely related minor peaks obtained were shown at 3551, 2516, 1347, 1104, 1028 and 813 cm^− 1 for PaL-AgNPs and 3536, 2509, 1741, 1361, 976 and 720 cm^− 1 for PaR-AgNPs. These peaks can confirm that alkenes (= C-H-), esters (C = O), amides (N-H), and aliphatic amines (C-N) were responsible for the synthesis, capping and stabilization of the AgNPs. Earlier, Lateef et al. [31] reported that phenolics and proteins were responsible for the bio-fabrication of AgNPs by the leaf extract of Petiveria alliacea.

The results from TEM analysis (Fig. 4) clearly showed that the particles of both PaL-AgNPs and PaR-AgNPs were spherical with the presence of silver. While PaL-AgNPs ranged in size from 5.95–28.91 nm, the PaR-AgNPs were 11.89–76.19 nm. These observations concur with Lateef et al. (2018) that reported the spherical AgNPs biosynthesized by the leaf of P. alliacea.

3.2 Fumigant toxicity effects of insecticides on adult cockroaches

As presented in Table 1, the applied treatments exhibited significant control of adult cockroaches. Lambdachyalothrin killed 100% of cockroaches after 1 h of exposure, while PaL-AgNPs applied at 10% v/v killed 33.3% of cockroaches and 50% of the cockroaches were killed by PaR-AgNPs 10% v/v. After 2 h of exposure, 50 and 66.7% mortality was observed in the container treated with PaR-Ag and PaL-AgNPs respectively applied at 10%, while no mortality was observed in the container treated with Pa-AgNPs applied at 1% and 5% v/v. However, applied PaR-AgNPs and PaL-AgNPs at 10% v/v resulted in 100% mortality after 3 h of application meanwhile, PaR-AgNPs at 5% killed higher number of the studied insect at 3–6 h (41.7, 66.7 and 91.7 respectively) than PaL-AgNPs applied at 5%v/v at the same hours of exposure.

After 4 h of application, PaR-AgNPs at 1% caused higher significant mortality (25%) than PaL-AgNPs applied at the same rate. There was an increase in the mortality of cockroaches treated with PaL-AgNPs at 5% v/v after 16 h and 100% mortality of the cockroaches was recorded at 16 h by PaR-AgNPs and PaR-AgNPs at 5% v/v.

Table 1

Effects of Insecticides on Cockroaches (Fumigant Toxicity)
		Hours of exposure
Treatments	Rate	1	2	3	4	6	16	20
Lambdacyhalothrin		100.0^a	100.0^a	100.0^a	100.0^a	100.0^a	100.0^a	100.0^a
Control		0.0^d	0.0^c	0.0^c	0.0^d	0.0^d	0.0^c	0.0^c
PaL-AgNPs	1	0.0^d	0.0^c	0.0^c	0.0^d	8.3^cd	16.7^c	33.3^c
	5	0.0^d	0.0^c	33.3^b	41.7^c	75.0^b	100.0^a	100.0^a
	10	33.3^c	66.7^b	100.0^a	100.0^a	100.0^a	100.0^a	100.0^a
PaR-AgNPs	1	0.0^d	0.0^c	8.3^c	25.0^cd	25.0^c	58.3^b	58.3^b
	5	0.0^d	0.0^c	41.7^b	66.7^b	91.7^ab	100.0^a	100.0^a
	10	50.0^b	50.0^b	100.0^a	100.0^a	100.0^a	100.0^a	100.0^a
Rates	1	25.0^b	27.1^b	27.1^c	31.3^c	35.4^b	41.7^b	48.0^b
	5	18.8^c	25.0^b	43.8^b	52.1^b	66.7^a	75.0^a	75.0^a
	10	45.8^a	54.2^a	75.0^a	75.0^a	75.0^a	75.0^a	75.0^a

Means with the same superscript(s) are significantly the same at 5% probability

3.3 Contact toxicity effects of insecticides on adult cockroaches

Table 2 shows the effect of contact toxicity of biosynthesized AgNPs on adult cockroaches. Application of PaR-AgNPs and PaL-AgNPs at 10% had the same significant effects on the cockroaches after 1 h of exposure meanwhile no mortality was recorded on the cockroaches treated with PaR-AgNPs and PaL-AgNPs at 1% and 5% v/v within 1 h of application. After 2 h of exposure, 93.3% of cockroaches were dead in lambdacyhalothrin treated containers while 50% of mortality was observed on the cockroaches treated with PaR-AgNPs and PaL-AgNPs at 10% v/v and 8.3% of cockroaches were found dead on the container treated PaL-AgNPs at 5% v/v but no mortality on the cage treated with PaR-AgNPs at 5%. 100% mortality was recorded in the lambdacyhalothrin treated containers while PaR-AgNPs and PaL-AgNPs applied at 10% killed 75% cockroaches after 3 hours meanwhile 100% mortality was achieved after 4 hours of exposure by the tested biosynthesized AgNPs at 10%. Throughout the experimental trials, none of the treated cockroaches by the PaR-AgNPs and PaL-AgNPs 1% v/v was found dead.

Table 2

Effects of Insecticides on Cockroaches (Contact Toxicity)
		Hours of Exposure
Treatments	Rate	1	2	3	4	6	16	20
Lambdacyhalothrin		80.0^a	93.3^a	100.0^a	100.0^a	100.0^a	100.0^a	100.0^a
Control		0.0^c	0.0^d	0.0^e	0.0^d	0.0	0.0	0.0
PaL-AgNPs	1	0.0^c	0.0^d	0.0^e	0.0^d	0.0^d	0.0^d	0.0^d
	5	0.0^c	8.3^c	16.7^d	16.7^c	16.7^c	16.7^c	33.3^c
	10	25.0^b	50.0^b	75.0^b	100.0^a	100.0^a	100.0^a	100.0^a
PaR-AgNPs	1	0.0^c	0.0^d	0.0^e	0.0^d	0.0^d	0.0^d	0.0^d
	5	0.0^c	0.0^d	50.0^c	58.3^b	58.3^b	58.3^b	58.3^b
	10	25.0^b	50.0^b	75.0^b	100.0^a	100.0^a	100.0^a	100.0^a
Rates	1	32.5^a	48.3^a	62.5^a	75.0^a	75.0^a	75.0^a	75.0^a
	5	20.0^b	27.5^b	43.8^b	43.8^b	43.8^b	43.8^b	47.9^b
	10	20.0^b	23.3^b	25.0^c	25.0^c	25.0^c	25.0^c	25.0^c

Means with the same superscript(s) are significantly the same at 5% probability

Control of household insect pests with synthetic insecticides is not really encouraging due to their environmental negative effects and high cost of procurement. Use of nanoparticles in the control of agricultural and household pests has been widely advocated due to their environmental friendly nature and their high entomotoxic effects [31, 32]. The root and leaf extracts of P. alliacea bio-fabricated nearly spherical AgNPs, having size range of 5.95–76.91 nm within 10 minutes of reaction. Earlier, Lateef et al. [31] reported the use of leaf aqueous extract of P. alliacea for the synthesis of AgNPs with antimicrobial, anticoagulant and antioxidant activities. However, there is no report on the use of the root extract of the plant for the synthesis of AgNPs until now. Also, this is the first report of the evaluation of biogenic AgNPs synthesized by P. alliacea as insecticides against cockroaches. Therefore, Pa-AgNPs were tested on common household insect (cockroach) with the two modes of application (contact and fumigant) at three concentrations (1, 5 and 10% v/v).

The results revealed that biosynthesized AgNPs exhibited insecticidal action against the studied insect, which clearly suggested that the tested Pa-AgNPs had insecticidal properties. This observation aligns with Yeguerman et al. [33] that reported the significant effects of biosynthesized polymeric nanoparticles from peppermint, Palmarosa geranium, lavender and rosemary on German cockroach. However, the extracts of P. alliacea used in the formulation of insecticides had been reported in some studies. For instance, the oil extracted from P. alliacea reportedly killed adult mosquitos within 2 h of application [34]. Similarly, P. alliacea and some other plant extracts had been tested against field insect pests of okra and cowpea [35, 36]. However, the existing studies have shown that the effectiveness of P. alliacea used in the formulation of nano-insecticides can be attributed to the organic compound called dibenzyltrisulfide [37, 38]. Insecticidal effect of this Pa-AgNPs was made possible due to the ability of nanoparticles to bind and adsorb other chemical compounds with ease and circulate them in insect body systems [39, 40]. In addition to this, Azeez et al. [41] and Lateef et al. [30] reported that AgNPs had high penetration potential of the particles of cellular metabolism which might cause deformation of DNA and inhibits the normal functioning of enzymes of the target organisms. Also, nanoparticles have been reported to have had effects on insects’ antioxidant and inhibited enzymes which eventually results to their death [42].

Comparatively, there was no significant difference in the efficacy of P. alliacea leaf and root in the formulations of nano-insecticides which indicates that any of this plant part can be used in the formulation. Within one hour of application, just only 25% of the studied cockroaches were killed through contact, while 50% mortality was observed through fumigant exposure at highest concentrations. Similarly, 100% mortality was recorded within 3 h of exposure through fumigant action. Meanwhile, it took 4 h to achieve 100% mortality through contact action. This is an indication that the route of exposure of Pa-AgNPs is one of the major determinants in the control of cockroaches. However, delayed insecticidal effects observed through contact toxicity might have been linked to the inability of nano-insecticide formulations to penetrate cuticle of the cockroaches easily and this suggests that Pa-AgNPs killed faster through respiratory system. This observation substantiates Abdel-Raheem and Eldafrawy [43] that reported that silver nanoparticles tested on the cockroaches through fumigant killed faster than the contact toxicity. Earlier study has proved that essential oil nanoparticles exhibited insecticidal action through contact against German cockroach [44]. This implies that the route of exposure is one of the factors influenced the toxicity of nano-insecticides formulations.

Data collected also suggested that the effectiveness of the tested biosynthesized AgNPs depended on the concentrations applied irrespective of mode of application. For instance, 75% mortality of cockroach was recorded at 10% v/v, while 43.8–52.1% mortality was achieved at 5% v/v. Meanwhile, 25-31.3% of the tested cockroach was killed at 1% after 3 h of application. This present study concurs with the earlier observation by Abdel-Raheem and Eldafrawy [43] which reported that effectiveness of silver nanoparticles against cockroaches was dose-dependent.

Although synthetic insecticide (lambdacyhalothrin) killed faster than biosynthesized AgNPs formulations, this is an indication that the tested nano-insecticides had delayed effects on the studied insect and this action is one of the major characteristic features of botanical insecticides. However, biosynthesized nanoparticles have been described as environmental-friendly due to their less toxicity to humans and environment [31, 32]. Thus, the delayed effects of biosynthesized AgNPs against cockroaches cannot be an absolute obstacle to their use as an alternative to synthetic insecticides putting into the consideration, the environmental positive effects of nano-insecticides over synthetic insecticides.

Based on these findings, the use of P. alliacea mediated silver nanoparticles was found to be effective in the control of cockroaches. Also, either the leaf or root of P. alliacea can be used in the formulation of nano-insecticides. These nano-insecticide formulations act as contact and fumigant insecticides, though fumigant toxicity killed faster than contact toxicity. Application at highest concentration of 100% proved to be more effective than the other tested concentrations. The use of this AgNPs against household insects will be another means of protecting our environment against toxic chemicals and reduction in the over dependent on synthetic insecticides will be achieved. The formulation from P. alliacea can be a welcome development in the management of household pests especially cockroaches. Therefore, the result obtained is an eye opener due to the fact that biosynthesized AgNPs using P. alliacea can be developed and commercialized as industrial products in the nearest future.

Ethics approval and consent to participate: Not applicable

Consent for publication: Not applicable

Availability of data and material: All data generated or analysed during this study are included in this published article [and its supplementary information files].

Competing interests: The authors declare that they have no competing interests.

Funding: This research did not receive any funding.

Acknowledgements: Not applicable.

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No competing interests reported.

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Anti-cockroach Activities of Biosynthesized Silver Nanoparticles using Petiveria alliacea Extracts

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Abstract

Background

Methods

Results

Conclusion

Figures

Highlights

1 Background

2 Methods

2.1 Materials

2.2 Test Insect Collection

2.3 Biosynthesis and Characterization of Silver Nanoparticles

2.4 Bioassay

2.4.1 Contact toxicity bioassay

2.4.2 Fumigant Toxicity bioassay

2.5 Data collection and statistical analysis

3 Results

3.1 Biosynthesis and Characterization of AgNPs

3.2 Fumigant toxicity effects of insecticides on adult cockroaches

3.3 Contact toxicity effects of insecticides on adult cockroaches

4 Discussion

5 Conclusion

Declarations

References

Additional Declarations

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