Agricultural Research Management Information System

Home
Research Summary
Search
About Us
- ARMIS
- Brochure
Contact Us
Report
User Request
Data Input
Help
- Operation Manual
  - PDF
  - Video
- Program Area & Commodity
We have reached 37600 number of research entries at this moment.

- Logout

Research Detail

Home
Research
Detail

Thin Layer Chromatographic Detection of Enrofloxacin Antibiotic Residues in Poultry Tissues

Md Mominul Islam Bhuiyan
Department of Pharmacology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Md Shafiqul Islam
Department of Pharmacology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Md Rakibul Hasan
Department of Pharmacology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Kazi Rafiqul Islam
Department of Pharmacology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Abstract/ Executive Summary:

Antibiotic residues remain in the edible portions of meat animals that have been treated with antibiotics. The aim of this study was to detect enrofloxacin residue after discriminate and indiscriminate administration and investigate the effect of enrofloxacin in the growth of poultry. 18 broilers DOC (Cobb-500) were collected & reared for up to 31 days. On day 16, they were randomly divided into 3 groups, namely Group–A (Control group), Group- B (Discriminate group), and Group-C (Indiscriminate group). Each group contains 6 birds. The discriminate and indiscriminate groups were treated with antibiotics, enrofloxacin. In Group-B withdrawal period was followed and treatment was stopped before 7 days of sacrifice. On the other hand, the withdrawal period was not maintained in indiscriminate groups and antibiotic treatment was continued until the day of sacrifice. Bodyweight was recorded daily in the morning. On 31^st day mean body weight was highest in Group-C (1901.17 ± 15.22gm) and the lowest body weight was in Group-A (1453.33 ± 26.39gm). The differences among mean weight gain were statistically significant (P<0.005) in both discriminate & indiscriminate groups compared to the control group. Test results found in TLC showed that indiscriminate antibiotic group (Group-B) 50% liver samples, 33.33% kidney and 16.67% fat samples were enrofloxacin positive. No sample of thigh muscle, breast muscle, and spleen was positive. In the indiscriminate antibiotic group (group-C) all the samples were positive in the case of liver, kidney, fat, and spleen samples. Only 33.33% and 16.67 % samples were positive in the case of fat and thigh muscle samples. All the samples of the control group (Group-A) were negative. Overall, the present study documented the widespread abuse of enrofloxacin, and failure to implement the recommended withdrawal period will undeniably lead to deposition of residues in broiler tissues.

Keywords: Antibiotic residue; Enrofloxacin; Broiler; TLC

Location: Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Start Date:

End Date:

Program Area: Animal Health and Management

Commodity: Poultry

Objectives:

The aim of this study was to detect enrofloxacin residue after discriminate and indiscriminate administration and investigate the effect of enrofloxacin in growth of poultry.

Methodology:

The experimental broilers were used ethically and at the end of the experiment sacrificed humanely following the ethical and welfare guidelines set by the Animal Welfare and Experimental Ethics Committee of Bangladesh Agricultural University [approval 5 number: AWEEC/ BAU/2021(09)]. 18 apparently healthy day-old “Cobb-500” broiler chicks were purchased from CP Hatchery Ltd, Valuka, Mymensingh. . On the 16^th days of age, chicks were randomly divided into three groups (Group A, B & C). Each group contains 6 birds. The birds of Group-A, B, and C were kept in different cages. Group A was kept as untreated control & received non-medicated water. Groups B & C were administered with enrofloxacin at recommended therapeutic dose @10 mg/Kg, through drinking water. After 7 days, at the age of day 23; antibiotic supply was stopped in group B and the withdrawal period was maintained. In group C the antibiotic supply was continued until the day of sacrifice. Birds received their freshly prepared daily medication in the morning hour of each day. The concentration of enrofloxacin in the water to give the required dose per kilogram of body weight was calculated by determining the water consumption and body weight of each bird on the day of medication. At the end of the experiment six birds from each group were sacrificed ethically. Liver, kidney, breast muscle, thigh muscle, fat, and spleen samples were collected. Immediately after the collection of samples, these were washed individually several times in physiological saline to remove clotted blood and debris. All samples were marked separately and preserved at -20º C in polythene zipper bags for their extraction and analysis. The samples were stored in the deep freeze at −20°C until further advanced procedures were performed. Samples were grinded with a mortar & pastel properly. These samples were taken into properly cleaned and sterilized petridishes with proper care. From this 4g of sample was taken into the beaker with the help of electric balance and spatula. The homogenization was done with the addition of 10 ml phosphate buffer (pH-7.2). After proper mixing, protein contents of these samples were precipitated with the addition of 2 ml trichloroacetic acid (30%) maintaining sufficient care and attention. Then these samples were taken into properly cleaned and sterilized centrifuge tubes for centrifugation. The centrifugation was performed @60000 rpm for 20 min with the help of an automatically time-regulated centrifuge machine. Then the supernatant was collected in a new tube. The supernatant was extracted with an equal volume of diethyl ether to perform de-fatation. Then the mixture was kept for 10 min to become into a separate layer. Then these mixtures were separated from each other, and the upper oily layer was discarded but only the bottom layer was collected. This extraction of supernatant was repeated twice with diethyl ether. Then, the extracts were collected into a screw cap vial with proper care and kept in a refrigerator for further advanced analysis. The total procedure was performed as the reference cited by Poppelka et al., 2005. TLC plate (MN-Germany), TLC tank, and UV detection box (UV light: F18W-Germany) were used. TLC was performed according to Tajick and Shohreh, 2006; Islam et al., 2021; Ali et al., 2020; Das et al., 2020 with some required adjustments. TLC plate was cut into appropriate size (4x5 cm) from 20x20 cm. A straight line was drawn across the plate approximately 2 cm from the bottom by a pencil. Another straight line was drawn across the plate below 1 cm from the upper edge of the plate. Desired spots marking were marked on the bottom line where analytes were dropped. Spots were applied to the plate using thin capillary glass pipettes. A volume of 50 μl was used for spotting. The plate was placed in a TLC tank (contained mobile phase; Butanol: distilled water: acetic acid = 60:20:20) and covered by a lid and it was left until the mobile phase reached the upper line. Spots were visualized in a UV detection box at 256 nm. Spots marking were done by pencil for calculation of retention factor (Rf). These measurements are the distance traveled by the solvent, and the distance traveled by individual sample spots. Same Rf value of standard and sample considered similar compound. Experimental data were introduced and stored in Microsoft Excel-2010 and results were analyzed.

Source of Information: Asian Australas. J. Food Saf. Secur. 2021, 5 (1), 11-18

Article Link (Online Journal): doi: 10.3329/aajfss.v5i1.55013

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Fluoroquinolones constitute an expanding group of synthetic antibiotics, widely used in the treatment of infections in both human and veterinary medicine. A number of these drugs have been licensed to be administered in broiler chickens for the prophylaxis and treatment of respiratory, renal and digestive infections in different regions of the world. Fluoroquinolones are one of the few classes of antimicrobial agents with activity against the full range of pathogens involved in broiler chickens, such as Campylobacter jejuni, Salmonella, Shigella, or Escherichia coli, is commonly used. Currently, the widespread use of these antimicrobials such as enrofloxacin in the poultry industry has become a matter of concern because it has led to the emergence of resistance in Salmonella serovars, Campylobacter spp. and E. coli . This situation raises public health concerns regarding the reduction in the clinical efficacy of fluoroquinolones in human medicine. In addition, the use of fluoroquinolones in food-producing animals may leave drug residues in foods. These residues represent a risk to public health, including stimulation of bacterial resistance, alterations on intestinal microflora, and hypersensitivity reactions. Extra label use of these drugs or unintentional contamination of feed for poultry (cross-contamination during premix manufacture or during feed transport) may be the source of violative drug residues in meat for human consumption. Therefore, the depletion of these drugs in meat and other edible tissues should be assessed. Considering the above-mentioned issues and the fact that published information regarding enrofloxacin depletion in poultry meat and offal is scarce the present study was designed. Administration of enrofloxacin was started at 16 days of age and continued for 31^st days of age. Mean body weight was highest for an indiscriminate group. The mean differences among the three groups were statistically significant (<0.05). The TLC was performed for the identification of antibiotic residues. The analysis revealed that all the samples were positive in an indiscriminate group with an exception of fat tissue (66.7%). There was no positive sample in the control group. The results indicate the presence of residues, whereas the usage of this contaminated meat causes resistance in consumers and seems to be a public health threat. Thus, there is a need to educate the farmers about the ill effects of residual drugs on human health and withdrawal time in poultry birds. National authorities should also adopt more judicious approaches to ensure prudent use of antibiotics in food animals.

Knowledge Management: Journal