Generic selectors
Exact matches only
Search in title
Search in content
Post Type Selectors
Filter by Categories
Case Report
Case Report, Medical Science and Practice
Case Report, Neurosciences and Behavioral Sciences
Commentaries, Neurosciences and Behavioural Sciences
Drug Evaluation and Clinical Trials, Original Research Article
Editorial
Epidemiology and Public Health, Original Research Article
Ethnomedicine and Phytomedicines, Original Research Article
Ethnomedicine and Phytomedicines, Review Article
Medical Science and Practice, Case Report
Medical Science and Practice, Original Research Article
Minireview Pharmacology and Toxicology
Minireview, Pharmacy Profession and Advocacy
Miniview, Pharmaceutical Education
Original Research Article, Drug Evaluation and Clinical Trials
Original Research Article, Drug Research and Development
Original Research Article, Ethnomedicine and Phytomedicines
Original Research Article, Neurology
Original Research Article, Pharmaceutical Education
Original Research Article, Pharmaceutics and Drug Delivery
Original Research Article, Pharmacotherapy/Pharmaceutical Care
Original Research Article, Pharmacy Practice
Pharmaceutical Technology and Manufacturing, Original Research Article
Pharmacology and Toxicology, Original Research Article
Pharmacotherapy/Pharmaceutical Care, Case Report
Pharmacotherapy/Pharmaceutical Care, Original Research Article
Pharmacy Practice, Original Research Article
Pharmacy Profession and Advocacy, Original Research Article
Review Article
Review Article, Medical Devices
Review Article, Perspective
Review Article, Pharmacotherapy/Pharmaceutical Care
Review Article, Pharmacy Practice
Generic selectors
Exact matches only
Search in title
Search in content
Post Type Selectors
Filter by Categories
Case Report
Case Report, Medical Science and Practice
Case Report, Neurosciences and Behavioral Sciences
Commentaries, Neurosciences and Behavioural Sciences
Drug Evaluation and Clinical Trials, Original Research Article
Editorial
Epidemiology and Public Health, Original Research Article
Ethnomedicine and Phytomedicines, Original Research Article
Ethnomedicine and Phytomedicines, Review Article
Medical Science and Practice, Case Report
Medical Science and Practice, Original Research Article
Minireview Pharmacology and Toxicology
Minireview, Pharmacy Profession and Advocacy
Miniview, Pharmaceutical Education
Original Research Article, Drug Evaluation and Clinical Trials
Original Research Article, Drug Research and Development
Original Research Article, Ethnomedicine and Phytomedicines
Original Research Article, Neurology
Original Research Article, Pharmaceutical Education
Original Research Article, Pharmaceutics and Drug Delivery
Original Research Article, Pharmacotherapy/Pharmaceutical Care
Original Research Article, Pharmacy Practice
Pharmaceutical Technology and Manufacturing, Original Research Article
Pharmacology and Toxicology, Original Research Article
Pharmacotherapy/Pharmaceutical Care, Case Report
Pharmacotherapy/Pharmaceutical Care, Original Research Article
Pharmacy Practice, Original Research Article
Pharmacy Profession and Advocacy, Original Research Article
Review Article
Review Article, Medical Devices
Review Article, Perspective
Review Article, Pharmacotherapy/Pharmaceutical Care
Review Article, Pharmacy Practice
View/Download PDF

Translate this page into:

Original Research Article
Drug Research and Development
2024
:3;
5
doi:
10.25259/AJPPS_2024_005

Antibacterial potentials of extracts from Gryllotalpa gryllotalpa, Pentodon algerinum grubs, and Gypsonoma euphraticana larva frass

Department of Biology, University of Mosul, Iraq.

*Corresponding author: Atallah Fahad Mekhlif, PhD Department of Biology, University of Mosul, Mosul Iraq. prof.atallah@uomosul.edu.iq

Licence
This is an open-access article distributed under the terms of the Creative Commons Attribution-Non Commercial-Share Alike 4.0 License, which allows others to remix, transform, and build upon the work non-commercially, as long as the author is credited and the new creations are licensed under the identical terms.

How to cite this article: Bader MF, Mekhlif AF. Antibacterial potentials of extracts from Gryllotalpa gryllotalpa, Pentodon algerinum grubs, and Gypsonoma euphraticana larva frass. Am J Pharmacother Pharm Sci 2024:5.

Abstract

Objectives:

The overuse and abuse of antibiotics have accelerated antibiotic resistance, and to solve this problem, it has been found that many insect species have potential antimicrobial properties against a wide range of resistant pathogens. Our study tests the antibacterial activity of microbial defensive compounds included in body extract of insects inhabiting contaminated environments and frass of phytophagous insects.

Materials and Methods:

Through sequential extraction method by acidic methanol, chloroform, and hexane solvents, insect body extract of Gryllotalpa gryllotalpa, grubs of Pentodon algerinum besides feces of Gypsonoma euphraticana larvae were tested against Gram-positives Bacillus cereus, Bacillus coagulans, Staphylococcus aureus, and Salmonella typhi, Escherichia coli, and Klebsiella pneumoniae. The antibiotics ceftriaxone (CRO) and ampicillin (AM) were used as standard drugs. The antibacterial growth inhibition was estimated by well diffusion methods.

Results:

High significant antibacterial activity against the tested bacteria by acidic methanol then chloroform extracts, while hexane extract of all the three insect species only produced significant growth inhibition of S. aureus. In addition, growth inhibition 20.0 mm or more was induced by: MeOH extracts of G. gryllotalpa and P. algerinum for S. typhi and E. coli, besides chloroform G. gryllotalpa extract for S. typhi. The tested bacteria S. aureus, S. typhi, and K. pneumoniae were AM-resistant, while E. coli was both AM and CRO-resistant.

Conclusion:

Acidic meOH and chloroform body extract of G. gryllotalpa and P. algerinum and larvae G. euphraticana feces extract possess bioactive compounds with promising antibacterial properties, for overcoming antibiotic resistance.

Keywords

Insect extracts
antibacterial
antibiotic resistance
Gryllotalpa gryllotalpa
Gypsonoma euphraticana
Pentodon algerinum

INTRODUCTION

The beneficial uses of most insects relate to honey and edible insects as food, silk for clothing, pollinator insects for plant pollination, and few traditional medicinal applications, but little is known about developing potential drugs from insect bodies depending on their innate immunity properties as reservoirs of antimicrobial agents. The overuse of antibiotics since the last decade of the 20th century has led to the emergence of antibiotic resistance.[1] Moreover, many pathogenic bacteria acquire resistance to more than one antibiotic, which so referred to as multidrug resistance, some of them are even resistant to any known antibiotics and so named pan-drug resistance.[2,3] Now, drug resistance is one of the 10 problems that threaten the world,[4,5] with annual proportional increasing resistance of the fatal pathogenic species to present antibiotics.[6] Today, drug resistance encourages searching for new alternative resources. One of these resources deals with the insect world, which aims to separate active antibacterial ingredients as templates for a new generation of the drug industry. Most studies in this field were first emphasized as survey study on the insect body extracts,[7-9] or bacterial inhibition by parts of the insect.[10-13] In more advanced studies, peptides with low molecular weights had been identified, and their growth inhibition activity was tested against a wide spectrum of Gram-negative and Gram-positive pathogenic bacteria. Therefore, many active metabolic compounds were separated and identified, with promising bacteria growth inhibition.[14,15] Moreover, many of the present drug-resistant bacteria are sensitive to insect antimicrobial peptides,[16-19] or epicuticular content lipids of the exoskeleton,[20-22] with promising results. Despite the huge diversity of the insect taxa, there has been slow progress in insect therapeutics, for instance, melittin from bees and alloferon from blow flies.[23,24]

In the light of the adaptation hypothesis, insects in polluted habitats have evolved high antimicrobial defense mechanisms. On this scope, the insect body extracts of the imago mole cricket, Gryllotalpa gryllotalpa, scarab beetle Pentodon algerinum grubs, and feces the leaf silk-webbing Gypsonoma euphraticana inhabited the host plant Populus euphratica were tested on the growth inhibition in vitro the pathogenic bacteria; Bacillus cereus, Bacillus coagulans, Staphylococcus aureus, Salmonella typhi, Escherichia coli, and Klebsiella pneumoniae.

MATERIALS AND METHODS

Insects

The tested insects were reared from their native environment in Mosul province/Iraq (36° 22'35 43° 08'32" E). Mole cricket G. gryllotalpa was collected manually from the house garden infested with the pest around a light source in the rainy season. Specimens of the scarab grubs, P. algerinum (about 30 mm long) were picked up from the earthen cells in a depth of about 30 centimeters the last spring. Feces were removed from the Populus Euphratica leaves housing the G. euphraticana.

Bacteria isolates

The human pathogenic bacteria had been used as references for evaluating in vitro antibacterial activity of the insect extracts. The Gram-positives are B. cereus, B. coagulans, and S. aureus, while S. typhi, E. coli, and Klebsiella pneumoniae are Gram-negatives. Bacteria isolates were identified and brought from the Microbiology laboratory/Department of Biology/College of Education for Pure Sciences/Mosul University/Iraq.

Culture media

The culture growth media, Muller–Hinton agar from NEOGEN Culture Media (foodstafety.neogen.com) had been purchased.

Extraction solvents

The insect body extracts were prepared using the following polar solvents with descending polarity indices values; water (10.2), dimethyl sulfoxide DMSO (7.2), acetic acid (6.0), methanol (5.1), chloroform (4.1), and hexane (0.1).

Bacteria isolation

Each of the bacteria species was inoculated on a new nutrient agar plate by loop full bacteria and then incubated for 24 h. at 37°C to obtain an active cultivar. The prepared plates were used either for experimental testing or kept at 4°C as stock inoculums for subsequent experiments.

Insect crude extracts

The mole crickets and scarab grubs were killed by lowering their temperatures in the refrigerator, then in the oven dried at 35°C. 100 g of dried insects and 25 g of larval feces were grounded by an electric mill, sequential separation of active constituents through a 3-stage solvent elution method which modified after.[7,25] The first step includes extraction by acidic methanol (90% meOH + 9% H2O + 1% CH3COOH) solvent, then the filtrate dried, and the precipitate secondly eluted by chloroform, and within the last (third) stage of the elution by hexane solvent. The three obtained dried extracts for each insect material were preserved at 4°C. For experimentation, the dried extract dissolved in DMSO, and the applied concentration for all the experimental treatments was 250 mg/mL.

Antibacterial susceptibility assay

Antibacterial activity was evaluated by the well diffusion method. The inhibition zones were recorded in millimeters (mm) using a ruler. Briefly, Muller–Hinton agar (MHA) plates were inoculated with the activated model bacteria isolates under aseptic conditions, and the wells (diameter = 8 mm) were filled by the test samples and incubated at 37 ºC for 24 h. Together, discs of standard drugs Ceftriaxone (CRO) and Ampicillin (AM) were fixed in MHA plates. The diameter of the clear growth to inhibition zones was measured. Inhibition rank was categorized according to Mohtar et al.[26] as follows: ≥8 mm (good), 6–7 mm (moderate), 4–5 mm (weak), and 2–3 mm (very weak).

Data analysis

All treatments were repeated in three replicates. The data was tabulated as means ± standard deviation. Mean differentiations at P ≤ 0.5 were conducted; using a one-way Analysis of Variance Duncan’s multiple range test.[27]

RESULTS

Antibacterial effect of the insect extracts

The present study deals with the antibacterial ability of the dry body ingredients of insects inhabiting polluted environments by means of growth inhibition zones of pathogenic bacteria. The antibacterial activity of body extracts of G. gryllotalpa, grubs of P. algerinum, and grounded feces of the leaves webbing moth, G. euphraticana, is shown in Tables 1-3. These extracts were prepared by sequential elution by gradual polarity indices of the applied solvents. The determined growth inhibition zone depended on the source of the extract and bacterium species.

Table 1: Antimicrobial activity of body extracts of mole cricket, Gryllotalpa gryllotalpa against pathogenic bacteria by inhibition of clear zone parameter.
Bacteria species Sequential solvents used
in extraction
Acidic meOH Chloroform Hexane
Bacillus cereus 19.0±0.0b 21.5±0.5a 16.0±0.0c
Bacillus coagulans 13.5±0.5a 12.0±1.0b 11.5±0.5b
Staphylococcus aureus 15.5±0.5b 16.8±0.8ab 18.0±1.0a
Salmonellatyphi 18.0±1.0b 21.7±0.8a 0.0±0.0c
Escherichia coli 20.5±0.5a 18.0±1.0c 0.0±0.0c
Klebsiella pneumoniae 19.0±1.0b 25.3±1.0a 0.0±0.0c

Horizontal means±standard deviations with different letters are significantly different at P≤0.05 (Duncan’s test)

Table 2: Growth inhibition zones (mm) of marker bacteria caused by fecal extract of moth larvae Gypsonoma euphraticana.
Bacteria species Sequential solvents used in extraction
Acidic meOH Chloroform Hexane
Bacillus cereus 16.2±0.3a 13.7±0.3b 10.7±0.6c
Bacillus coagulans 10.0±0.0b 12.2±2.5a 0.0±0.0c
Staphylococcus aureus 15.7±0.6b 15.20.3c 17.0±0.0a
Salmonella typhi 14.5±0.5a 10.5±0.5b 0.0±0.0c
Escherichia coli 11.7±0.6a 10.2±0.3b 0.0±0.0c
Klebsiella pneumoniae 15.5±0.5a 10.8±0.3b 9.8±0.3c

Horizontal means±standard deviations with different letters are significantly different at P≤0.05 (Duncan’s test)

Table 3: Antimicrobial activity of body extracts of white grub larvae, Pentodon algerinum extract represented by growth clear zones.
Bacteria species Sequential solvents used in extraction
Acidic meOH Chloroform Hexane
Bacillus cereus 15.2±0.8a 10.8±0.3b 14.3±0.9a
Bacillus coagulans 13.7±0.6a 10.2±0.3b 10.8±0.3b
Staphylococcus aureus 11.7±0.6c 14.0±1.0b 17.7±0.6a
Salmonella typhi 21.8±0.8a 10.0±1.0c 13.7±0.6b
Escherichia coli 20.0±0.0a 7.0±0.0b 0.0±0.0c

Horizontal means±standard deviations with different letters are significantly different at P≤0.05 (Duncan’s test)

For G. gryllotalpa extract, Table 1 exhibits growth inhibition of all the testing Gram-positive bacteria (B. cereus, B. coagulans, and S. aureus) by the three applied polar solvents ranging from 21.5 mm (for B. cereus) to 12.0 mm (for chloroform extract). While, only acidic methanol and chloroform inhibited the growth of the treated Gram-negative bacteria; S. typhi, E. coli, and K. pneumoniae, with higher clear zones of 25.3 mm for K. pneumoniae at chloroform extract and lower growth inhibition zone 18.0 mm for S. typhi and E. coli at acidic methanol and chloroform extracts, respectively.

The fecal extract of moth larvae, G. euphraticana inhibiting all Gram-positive bacteria except hexane extract for B. coagulans, is evoked in Table 2. On the other hand, only K. pneumoniae from Gram-positive bacteria were inhibited by hexane extract with 9.8 mm.

The grub beetle, P. algerinum extract with all three polar solvents, inhibited growth of the Gram-positives which ranged between 17.7 mm for S. aureus by hexane and 10.2 mm for B. coagulans with chloroform extract [Table 3].

Growth inhibit ability at each solvent extract bacteria

Each of the Tables 1-3 revealed how long the growth inhibition zones obtained by the extracts of G. gryllotalpa, G. euphraticana, larva feces and grubs P. algerinum, which were separately prepared by the following solvents; acidic methanol (mixed solvents), chloroform, and hexane.

For acidic methanol extract: The clear zones between 18.0 and 20.5 mm are shown in Table 4 demonstrated by the action of G. gryllotalpa against S. aureus, B. cereus, K. pneumoniae, and E. coli. Besides, feces extract gave a diameter clear zone ranging from 14.5 to 16.2 mm for the bacteria S. typhi, S. aureus, K. pneumoniae, and B. cereus, respectively. The extract of the grub beetle P. algerinum inhibited the growth of S. aureus, K. pneumoniae, B. coagulans, and B. cereus, while 20.0 and 21.8 mm for E. coli and S. typhi, respectively.

Table 4: Antibacterial inhibition by acidic meOH body extracts G. gryllotalpa and fecal extract of the moth G. euphraticana and Scarab grub P. algerinum against the marker bacteria.
Insect extract The growth inhibition zone (mm) of the bacteria
B. cereus B. coagulans S. aureus S. typhi E. coli K. pneumoniae
G.gryllotalpa 19.0±0.0bA 13.5±0.5dC 15.5±0.5cB 18.0±1.0bC 20.5±0.5aA 19.0±1.0bB
G.euphraticana 16.2±0.3aB 10.0±0.0dD 15.7±0.6aB 14.5±0.5bD 11.7±0.6cB 15.5±0.5aC
P.algerinum 14.7±1.5cB 13.7±0.6cdC 11.7±0.6eC 21.8±0.8aB 20.0±0.0bA 12.7±0.6deD
CRO (ve+) 110.0±0.5cC 15.3±1.5bB 17.2±0.8bA 24.3±1.2aA 0.0±0.0dC 26.0±0.0aA
AM (ve+) 14.7±0.6bB 22.3±0.6aA 0.0±0.0cD 0.0±0.0cE 0.0±0.0cC 0.0±0.0cE

Horizontal means±standard deviations with different (small) letters are significantly different at P≤0.05 (Duncan’s test). Means that vertical different (capital) letters are significantly different at P≤0.05 (Duncan’s test). G. gryllotalpa: Gryllotalpa gryllotalpa, P. algerinum: Pentodon algerinum, G. euphraticana: Gypsonoma euphraticana, B. cereus: Bacillus cereus, B. coagulans: Bacillus coagulans, S. aureus: Staphylococcus aureus, S. typhi: Salmonella typhi, E. coli: Escherichia coli, K. pneumonia: Klebsiella pneumoniae. CRO: Ceftriaxone, AM: Ampicillin

The diameters of growth inhibition zones of the cultured plates treated with extracts of the second phase chloroform are shown in Table 5. For mole G. gryllotalpa extract, the growth inhibition zone is mostly between 12.0 and 18.0 mm, except for B. cereus and K. pneumoniae 21.5 and 25.3 mm, respectively. However, P. algerinum grub extract was less effective with a range of 7.0–14 mm for all the experimental bacteria.

Table 5: Antibacterial activity of Chloroform body extracts G. gryllotalpa and fecal extract of the moth G. euphraticana and Scarab grub P. algerinum against the marker bacteria.
Insect extract Growth inhibition zone (mm) of the bacteria
B. cereus B. coagulase S. aureus S. typhi E. coli K. pneumoniae
G.gryllotalpa 21.5±0.5bA 12.0±0.0dC 16.8±0.8cA 21.7±0.6bD 18.0±1.0cA 25.3±1.0aB
G.euphraticana 13.7±0.5bC 12.2±2.5ccD 15.2±0.3aB 10.5±0.5cdC 10.2±0.3dB 10.8±0.3cdB
P.algerinum 10.8±0.3cD 10.2±0.3cdD 14.0±0.0aC 10.0±1.0dC 7.0±0.0eC 12.0±0.0bB
CRO (ve+) 11.0±0.5cD 15.3±1.5bB 17.2±0.8bA 24.3±1.2aA 0.0±0.0dD 260±2.0aA
AM (ve+) 14.7±0.6bB 22.3±0.6aA 0.0±0.0cD 0.0±0.0cB 0.0±0.0cD 0.0±0.0cC

Horizontal means±standard deviations with different (small) letters are significantly different at P≤0.05 (Duncan’s test). Means with vertical different (capital) letters are significantly different at P≤0.05 (Duncan’s test). G. gryllotalpa: Gryllotalpa gryllotalpa, P. algerinum: Pentodon algerinum, G. euphraticana: Gypsonoma euphraticana, B. cereus: Bacillus cereus, B. coagulans: Bacillus coagulans, S. aureus: Staphylococcus aureus, S. typhi: Salmonella typhi, E. coli: Escherichia coli, K. pneumonia: Klebsiella pneumoniae. CRO: Ceftriaxone, AM: Ampicillin

The antibacterial sensitivity variation between the marker bacteria treatment with the third (last) elution phase by hexane is illustrated in Table 6. Except for the bacteria, B. cereus, B. coagulans, and S. aureus were inhibited by extract G. gryllotalpa 16.0, 15.0, and 18.0 mm, respectively. Only, the bacteria B. cereus and S. aureus were affected by moth G. euphraticana larval frass with zones of inhibition 10.7 and 17.0 mm. It was found that E. coli resistant to grub P. algerinum hexane extract, and growth inhibition zones were determined (8.7, 13.8) for Gram-negative K. pneumoniae, and S. typhi, and 10.8, 14.3, and 17.7 mm for B. coagulans, B. cereus, and S. aureus, respectively.

Table 6: Antibacterial inhibition by Hexane extracts G. gryllotalpa and fecal extract of the moth G. euphraticana and Scarab grub P. algerinum against the pathogenic bacteria.
Insect extract Growth inhibition zone (mm) of the bacteria
B. cereus B. coagulans S. aureus S.typhi E. coli K. pneumoniae
G. gryllotalpa 16.0±0.0bA 11.5±0.5C 18.0±1.0aA 0.0±0.0cC 0.0±0.0dA 0.0±0.0dC
G. euphraticana 10.7±0.6bC 0.0±0.0cD 17.0±0.0aA 0.0±0.0cC 0.0±0.0cA 0.0±0.0cC
P. algerinum 14.3±1.0bB 10.8±0.3cC 17.7±0.6aA 13.8±0.6bB 0.0±0.0eA 8.7±0.6dB
CRO (ve+) 11.0±0.5cC 15.7±0.8bB 17.2±0.8bA 24.3±1.2aA 0.0±0.0dA 26.0±2.0aA
AM (ve+) 14.7±0.6bB 22.3±0.6aA 0.0±0.0cB 0.0±0.0cC 0.0±0.0cA 0.0±0.0cC

Horizontal means±standard deviation with different (small) letters is significantly different at P≤0.05 (Duncan’s test). Means with vertical different (capital) letters are significantly different at P≤0.05 (Duncan’s test). G. gryllotalpa: Gryllotalpa gryllotalpa, P. algerinum: Pentodon algerinum, G. euphraticana: Gypsonoma euphraticana, B. cereus: Bacillus cereus, B. coagulans: Bacillus coagulans, S. aureus: Staphylococcus aureus, S. typhi: Salmonella typhi, E. coli: Escherichia coli, K. pneumonia: Klebsiella pneumoniae. CRO: Ceftriaxone, AM: Ampicillin

Inhibition comparison between standard drugs and insect extracts

CRO caused antibacterial action (24.3, 26.0 mm) at treatment of the bacteria S. typhi and K. pneumoniae, and 11.0, 15.3, and 17.2 mm for B. cereus, B. coagulans, and S. aureus, respectively, but E. coli was not affected. The zones of inhibition by amoxicillin were restricted (14.7, 22.3 mm) with only B. cereus and B. coagulans, whereas the latters (S. aureus, S. typhi, and K. pneumoniae) were completely not responsive to the applied standard drugs [Table 4].

After testing with acidic meOH [Table 4] (with perpendicular columns); B. cereus was more inhibited (19.0, 16 2, and 15.0 mm) at G. gryllotalpa, G. euphraticana, and P. algerinum and then amoxicillin standard drugs (11.0 and 14.6 mm), respectively. The tested standard drugs were more effective than all the tested extracts. For S. aureus, their growth was inhibited with 17.2 mm by CRO (standard drug), whereas for G. gryllotalpa, G. euphraticana, and P. algerinum ranged between 16.5 and 11.6 mm, respectively. S. typhi was inhibited by CRO (24.3 mm) and the extracts were between 21.8 and 14.5 mm. Sensitivity of E. coli to the extracts was about 20.0 mm for G. gryllotalpa and P. algerinum and resistant to the standard drugs. K. pneumoniae was only inhibited by CRO (26.0 mm) and less with a range of 19.0–12.6 mm for the applied extracts.

Growth inhibition by chloroform extracts; zone diameters of B. ceseus with G. gryllotalpa and G. euphraticana extracts 21.5 and 13.7 mm, and less than (11.0, 14.7) for CRO and AM (+ve). In the case of B. coaculans, growth inhibition was 22.3 and 15.3 mm for the antibiotics(+ve) AM and CRO, and between 10.2 and 11.0 mm for the tested extracts. It was found only CRO inhibits the growth of S. aureus with near results for G. gryllotalpa and G. euphraticana extracts. The S. typhi is resistant to AM but sensitive (24.3 mm) to CRO 21.3 for G. gryllotalpa and 10.5 mm for both G. euphraticana, and P. algerinum extracts. E. coli and K. pneumoniae were resistant to the tested standard drugs except the second ones 26.0 mm with CRO, whereas growth inhibition by G. gryllotalpa, G. euphraticana, and P. algerinum (18.0, 10.0, and 7.0 mm) and (25.3, 10.8, and 12.0 mm) for E. coli and K. pneumoniae, respectively.

In comparison antibacterial treatment with hexane insect extracts with (standard drugs) CRO and AX: B. cereus inhibition (16.0 mm) with G. gryllotalpa more than that of the other two extracts, besides the antibiotics (11.0, 14.7 mm) CRO and AM. But (+ve) CRO and AM were more effective than tested insect extracts for B. coaculans. CRO had nearly the same antibacterial activity (17.2) with the applied G. gryllotalpa, G. euphraticana, and P. algerinum extracts against S. aureus. Only CRO had growth inhibition (24.3, 26.0 mm) to S. typhi and K. pneumoniae. However, E. coli is resistant to all antibiotics and insect extracts [Table 6].

DISCUSSION

Insects like other invertebrates have only innate immune system, therefore, have highly developed immune systems. Theoretically, because of their feeding habit and habitat like some other studied insects.[7,14,28,29] subterranean insects such as G. gryllotalpa, P. algerinum larvae, and webbing G. euphraticana larvae are in direct exposure to the pathogenic microbial agents. According to this hypothesis, our study gives promising results of potentially significant antibacterial properties. Due to overuse and abuse present antibiotics were led to overcoming annual antibiotic resistance to pathogenic and opportunistic bacteria. Insect body extracts and purified constituents from insect body parts were proven as one of the future antibiotics, and they took continuous interest by many alternative natural product researchers.[17,30,31] In the present study, the measured growth inhibition zone of any tested marked bacteria was related to the tested bacterium, source of the insect body extract, and polarity of the solvent used in extraction. Therefore, according to Mohtar et al.[26] susceptibility rank of the antibacterial agents, acidic meOH G. gryllotalpa extract had more significant activity (19.0 mm) for both B. coagulans and K. pneumoniae and 20.5 mm for E. coli [Table 1], while 5 of the 6 marked bacteria treated by G. euphraticana and P. algerinum were more significantly caused growth inhibition in relation to chloroform and hexane extracts, which ranged between good to moderate inhibition [Tables 1-3]. It was found qualitative and quantitative inhibition by chloroform after acidic methanol extracts through the sequential method so that only G. gryllotalpa extract caused growth inhibition between 21.5 and 25.3 mm for B. cereus, S. typhi and K. pneumoniae, and G. euphraticana and P. algerinum extracts were less than 15.2 mm for all the tested bacteria. On the other hand, the largest growth inhibition by hexane extract was 18.0 mm at S. aureus by G. gryllotalpa extract.

It is illustrated in Tables 1-3 that G. gryllotalpa extracted by all the three sequential polar solvents had more significant growth inhibition B. cereus than the standard drugs. It was found nearly the same effect of G. gryllotalpa extracted by all the solvents and CRO on S. aureus which is completely resistant to AM. Besides, equal moderate effect of all the applied extracts with hexane and CRO, and complete resistance to AM. S. typhi was inhibited by all the extracts, but less significant than CRO and resistant (0.0 mm) to AM. All the extracts had growth inhibition to E. coli, while at the same time had not responded to CRO and AM. K. pneumoniae was resistant to AM but sensitive (26.0 mm) to CRO which was better than the extracts, so all the extracts had significant inhibition in relation to AM.

CONCLUSION

In the present study, the good antibacterial activity of whole body extract of the insects that inhabiting polluted niches with pathogenic bacteria and other microbes, so the insect body reflex was represented by the production of antibiotic constituents. Therefore, a wide spectrum of Gram-positive and Gram-negative bacteria exhibited good sensitivity to body extract from subterranean G. gryllotalpa and grubs of P. algerinum and frass pellets of the confined living G. euphraticana larvae. Most of the extracts, especially acidic methanol have better activity than the (CRO and AM antibiotics) standard drugs.

Acknowledgment

Our gratitude goes to Mosul University authorities, the President of the university Prof. Dr. Kossay Alahmady for his scientific support, and the Head of the Biology Department; Prof. Dr. Mohmaad S. Faisal, for continued support.

Ethical approval

Institutional Review Board approval is not required.

Declaration of patient consent

Patient’s consent is not required as there are no patients in this study.

Conflicts of interest

There are no conflicts of interest.

Use of artificial intelligence (AI)-assisted technology for manuscript preparation

The authors confirm that there was no use of artificial intelligence (AI)-assisted technology for assisting in the writing or editing of the manuscript and no images were manipulated using AI.

Financial support and sponsorship

None.

References

  1. , , , et al. Evolution and emergence of antibiotic resistance in given ecosystems: Possible strategies for addressing the challenge of antibiotic resistance. Antibiotics. 2023;12:28. doi: 10.3390/antibiotics12010028
    [CrossRef] [PubMed] [Google Scholar]
  2. , , , et al. Multidrug-resistant, extensively drug-resistant and pan drug-resistant bacteria: An international expert proposal for interim standard definitions for acquired resistance. Clin Microbiol Infect. 2012;18:268-281. doi: 10.1111/j.1469-0691.2011.03570.x
    [CrossRef] [PubMed] [Google Scholar]
  3. , . Introduction: Antibiotic resistance. Chem Rev. 2005;105:391-394. doi: 10.1021/cr030100y
    [CrossRef] [PubMed] [Google Scholar]
  4. . Factsheet: Antimicrobial resistance. . Geneva: WHO; Available from: https://www.who.int/news-room/fact-sheets/detail/antimicrobial-resistance [Last accessed on 2023 Aug 03]
    [Google Scholar]
  5. . Global action plan on antimicrobial resistance. . Geneva: World Health Organization; Available from: http://apps.who.int/iris/bitstream/10665/193736/1/9789241509763_eng.pdf?ua=1 [Last accessed on 2023 Aug 03]
    [Google Scholar]
  6. . Global priority list of antibiotic-resistant bacteria, to guide research, discovery, and development of new antibiotics. . Geneva: WHO; Available from: https://remed.org/wp-content/uploads/2017/3 [Last accessed on 2023 Aug 03]
    [Google Scholar]
  7. , , , et al. Antimicrobial activity of 11 insects extracts against multi drug resistant (MDR) strains of bacteria and fungus. IOP Conf Ser Earth Environ Sci. 2019;252:022132. doi: 10.1088/1755-1315/252/2/022132
    [CrossRef] [Google Scholar]
  8. , , , et al. Crude extract of maggots: Antibacterial effects against Escherichia coli, underlying mechanisms, separation and purification. World J Gastroenterol. 2012;21:1510-1517. doi: 10.3748/wjg.v21.i5.1510
    [CrossRef] [PubMed] [Google Scholar]
  9. , , . Cockroaches, locusts, and envenoming arthropods: a promising source of antimicrobials. Iran J Basic Med Sci. 2018;21:873-877. doi: 10.22038/IJBMS.2018.30442.7339
    [CrossRef] [Google Scholar]
  10. , . Partial purification and characterization of antimicrobial peptide from the hemolymph of cockroach Periplaneta americana. J Appl Biol Biotechnol. 2020;8:6-11. doi: 10.7324/JABB.2020.80202
    [CrossRef] [Google Scholar]
  11. , , , et al. Evaluation of the antimicrobial activity of purified Spodoptera littoralis hemolymph against some pathogenic bacteria. Afr J Biol Sci. 2021;17:221-231. doi: 10.21608/AJBS.2021.195329
    [CrossRef] [Google Scholar]
  12. , . Yellow wasp Polistes flavus venom protein, its purification, solubilization and antimicrobial activity. Biomed J Sci Tech Res. 2017;1:154-158. doi: 10.26717/BJSTR.2017.01.000141
    [CrossRef] [Google Scholar]
  13. , , , et al. The antibacterial effect of American cockroach hemolymph on the nosocomial pathogenic bacteria. Avicenna J Clin Microb Infect. 2015;2:e23017. doi: 10.17795/ajcmi-23017
    [CrossRef] [Google Scholar]
  14. , , , et al. Separation of bioactive compounds from Haemolymph of scarab beetle Scarabaeus sacer (Coleoptera: Scarabaeidae) by GC-MS and determination of its antimicrobial activity. Int J Appl Biol. 2021;5:98-116. doi: 10.20956/ijab.v5i2.18539
    [CrossRef] [Google Scholar]
  15. , , , et al. Insect derived Laurica acid a promising alternative strategy to antibiotics in the antimicrobial resistance scenario. Front Microbiol. 2021;12:620798. doi: 10.3389/fmicb.2021.620798
    [CrossRef] [PubMed] [Google Scholar]
  16. , , , et al. New insect host defense peptides (HDP) from dung beetle (Coleoptera: Scarabaeidae) transcriptomes. J Insect Sci. 2021;21:12. doi: 10.1093/jisesa/ieab054
    [CrossRef] [PubMed] [Google Scholar]
  17. , . Antimicrobial peptides (AMPs): A promising class of antimicrobial compounds. J Appl Microbiol. 2022;132:1573-1596. doi: 10.1111/jam.15314
    [CrossRef] [PubMed] [Google Scholar]
  18. , , . Insect antimicrobial peptides, a mini review. Toxins. 2018;10:461. doi: 10.3390/toxins10110461
    [CrossRef] [PubMed] [Google Scholar]
  19. , , , et al. Insect antimicrobial peptides show potentiating functional interactions against Gram-negative bacteria. Proc Biol Sci. 2015;282:20150293. doi: 10.1098/rspb.2015.0293
    [CrossRef] [PubMed] [Google Scholar]
  20. , , . Characterization of antimicrobial peptides from local forest dwelling ants: In-vitro screening for antimicrobial activity. Eur J Biol Biotechnol. 2021;2:20-25. doi: 10.24018/ejbio.2021.2.1.138
    [CrossRef] [Google Scholar]
  21. , , , et al. Free fatty acids in the cuticular and internal lipids of Calliphora vomitoria and their atimicrobial activity. J Insect Physiol. 2013;59:416-429. doi: 10.1016/j.jinsphys.2013.02.001
    [CrossRef] [PubMed] [Google Scholar]
  22. , , , et al. Antimicrobial activity of untypical lipid compounds in the cuticular and internal lipids of four fly species. J Appl Microbiol. 2013;116:269-287. doi: 10.1111/jam.12370
    [CrossRef] [PubMed] [Google Scholar]
  23. , , . New alloferon analogues: Synthesis and antiviral properties. Chem Biol Drug Des. 2013;18:302-309. doi: 10.1111/cbdd.12020
    [CrossRef] [PubMed] [Google Scholar]
  24. , , , et al. Enhanced binding to and killing of hepatocellular carcinoma cells in vitro by melittin when linked with an oval targeting peptide screened from phage display. J Pept Sci. 2013;9:639-650. doi: 10.1002/psc.2542
    [CrossRef] [PubMed] [Google Scholar]
  25. , . Assessment an antibacterial activity of crud bodies, Ailolopus thaiossinus (Orthoptera) and Polistes watti larvae (Hymenoptera) by extracted cold and boiled solvents. J Entomol Zool Stud. 2022;10:62-67. doi: 10.22271/j.ento.2022.v10.i3a.9012
    [CrossRef] [Google Scholar]
  26. , , . Screening of novel acidified solvents for maximal antimicrobial peptide extraction from Zophobas morio Fabricius. Adv Environ Biol. 2014;8:803-809.
    [Google Scholar]
  27. . Multiple range and multiple F test. Biometric. 1955;11:1-42. doi: 10.2307/3001478
    [CrossRef] [Google Scholar]
  28. . In vitro screening the antibacterial activity of four whole body eusocial insects extracted by polar solvents. Intl J Mol Biol Biochem. 2021;3:19-24. doi: 10.33545/26646501.2021.v3.i1a.21
    [CrossRef] [Google Scholar]
  29. , . Antibacterial activity of secretion/excretion blow fly, Calliphora vomitoria (Diptera: Calliphoridae) third instar larvae in vitro. J Entomol Zool Stud. 2021;9:14-19.
    [Google Scholar]
  30. , , . Insect antimicrobial peptides: Advancements, enhancements and new challenges. Antibiotics. 2023;12:952. doi: 10.3390/antibiotics12060952
    [CrossRef] [PubMed] [Google Scholar]
  31. , , , et al. Antimicrobial peptides derived from insects offer a novel therapeutic option to combat biofilm: A review. Front Microbiol. 2021;12:661195. doi: 10.3389/fmicb.2021.661195
    [CrossRef] [PubMed] [Google Scholar]
Show Sections