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Therapeutic Potential of Plant Extracts Against Multidrug Resistance Poultry Bacteria

Nur-E-Alam
Department of Biotechnology and Genetic Engineering, Mawlana Bhashani Science and Technology University, Tangail -1902, Bangladesh

Md. Rayhan Ali
Department of Biotechnology and Genetic Engineering, Mawlana Bhashani Science and Technology University, Tangail -1902, Bangladesh

Md. Tarek Molla
Department of Biotechnology and Genetic Engineering, Mawlana Bhashani Science and Technology University, Tangail -1902, Bangladesh

Shahin Mahmud
Department of Biotechnology and Genetic Engineering, Mawlana Bhashani Science and Technology University, Tangail -1902, Bangladesh

Kaisar Ali Talukder
Department of Biotechnology and Genetic Engineering, Mawlana Bhashani Science and Technology University, Tangail -1902, Bangladesh

A. K. M. Mohiuddin
Department of Biotechnology and Genetic Engineering, Mawlana Bhashani Science and Technology University, Tangail -1902, Bangladesh

Abstract/ Executive Summary:

Plant extracts were evaluated on bacteria isolated from poultry farm for developing substitutive therapeutic agent of antibiotics. A diverse range of bacterial load observed both in total viable count (TVC) and in total coliform count (TCC) in 30 samples randomly collected from poultry feeds, drinking water and faeces. A total of six bacterial isolates e.g. Pseudomonas spp., Aeromonas spp., Citrobacter spp., Vibrio spp., Escherichia coli and Plesiomonas spp. were found in the samples cultured in MacConkey Agar medium. Fifteen antibiotics were studied against bacterial susceptibility. All the bacterial isolates exhibited multi -antibiotic resistance (MAR) with gross resistance to erythromycin and ampicillin. E. coli had the highest MAR (53.3%), and Vibrio spp. as well as Plesiomonas spp. both had the same MAR (46.7%). Methanolic extract of Terminalia chebula and Azadirachta indica showed significant zone of inhibition against all the tested bacteria. These finding s confirm the presence of multi drug resistant bacteria in poultry environment that reveals a possibility of cross-contamination to human and animals . The plant extracts could be developed into therapeutic drugs to rein antibiotic poultry resistant bacteria.

Keywords: Poultry, Antibiotic resistance, Plant extract, Cross-contamination, Safe poultry source

Location: Tangail district of Bangladesh

Start Date: 00-05-2019

End Date: 00-12-2019

Program Area: Animal Health and Management

Commodity: Poultry

Objectives:

The aim of this study was to show bacterial association in feeds, water, and chicken poultry faeces, their antibiotics susceptibility and utilization of some plant extracts to minimize the hazards and risk s related to bacterial contamination in poultry.

Methodology:

A total of 30 samples were collected from poultry practicing rural community in Tangail district of Bangladesh, of which 14 were poultry feed, 9 were water and 7 were faecal samples randomly collected from poultry stores and poultry farms between May and December, 2019. The feed samples were aseptically collected in sterile polyethylene bags while water and faecal materials were aseptically collected in sterile falcon tubes which were sealed and transported to the laboratory directly. A sterile warring blender was used to homogenize 1.0g of each feed and faecal sample into 10 ml of sterile distilled deionized water (Fawole and Oso 2001) resulting 1:10 dilution. Later on, serial dilutions up to 10^-6 for feed and faecal samples, and 10^-3 were prepared for water. For the determination of total viable count (TVC) and total coliform count (TCC), about 0.1ml of diluted samples were cultured in duplicate on nutrient agar and MacConkey agar media, respectively using pour plate method. These were incubated at 37ºC for 18hrs in an incubator. The results of TVC were expressed as the number of organism or colony -forming units per gram ( cfu/g) of feed and faeces samples and cfu/ml for water sample. The bacterial isolates were identified on the basis of morphological and biochemical tests such as Kligler iron agar (KIA) test, Motility -indole-urease (MIU) test and citrate utilization test (Holt et al. 1994). The susceptibility of the isolates against some common antibiotics and chemotherapeutics was tested using the Kirby -Bauer disk diffusion method on Mueller - Hinton agar (Hudzicki 2009, Watts et al. 2008). A total of 15 antibiotic discs (Oxoid, UK) such as ampicillin (10μg), azithromycin (15μg), ciprofloxacin (5μg), tetracycline (30μg), amoxicillin (10μg), nalidixic acid (30μg), and kanamycin (30μg), amikacin (30μg), chloramphenicol (30μg), erythromycin (15μg), gentamicin (10μg), levofloxacin (5μg), neomycin (30μg), norfloxacin (10μg), and tazobactam (110μg) were used. The interpretation of the antibiotic susceptibility was made according to CLSI (The Clinical and Laboratory Standards Institute 2014). Healthy and disease-free plant leaves of Azadirachta indica were collected directly from the plant and the dried ripe fruit of Terminalia chebula purchased from a local market of Tangail. The freshly collected leaves and dried ripe fruit s were washed with distilled water and dried in shade for two weeks and blended into powder using mortar. About 100g powder for both T. chebula ripe fruit and leaves of A. indica were extracted with 600ml of methanol (95% for T. chebula), and(30, 40 and 70% methanol for A. indica ) at 25°C for 48hrs (Sahreen et al . 2011). The crude extracts were filtered using Whatman No. 1 filter paper and evaporated by using a rotary evaporator. Antimicrobial activity of the plant extracts was tested using disc -diffusion method. Young culture of the test organisms in 2 ml sterile Mueller Hinton broth (MHB) were made from well isolated single colony obtained from 24hrs grown cultures. The test cultures were swabbed on the top of the solidified medium and allowed to dry. The disc containing plant extract (each disc contains 30 µl of plant extract in different methanolic concentration) were placed on the plate. The plates were then incubated at 37°C for 18hrs. The diameter of the zone of inhibition around each disc was measured in mm and the mean value was calculated (Hudzicki 2009). In this study, microbial were determined following standard formulae. Then the results were analyzed by SPSS ver . 20. Hierarchical analysis was used to estimate overall similarities of the bacterial resistance using their zones of inhibition. Statistical significance was set at a p<0.05. Microsoft Excel version 2016 was used to draw graphs wherever appropriate.

Source of Information: Plant Tissue Cult. & Biotech. 30(1): 119-130, 2020 (June)

Article Link (Online Journal): DOI: https://doi.org/10.3329/ptcb.v30i1.47797

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

These finding s confirm the presence of multi drug resistant bacteria in poultry environment that reveals a possibility of cross-contamination to human and animals. The plant extracts could be developed into therapeutic drugs to rein antibiotic poultry resistant bacteria.

Knowledge Management: Journal