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Residual Effect of Amoxicillin in Broiler

Mst. Shumiya Khatun
Department of Pharmacology, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

Dolan Das
Livestock Extension Officer
Department of Pharmacology, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh; LDDP, DLS, Natore, Bangladesh.

Mst. Sharmin Akter
Department of Pharmacology, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

Md. Ashraf Zaman Faruk
Department of Anatomy and Histology, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Sobrata Das
Department of Physiology, Sylhet Agricultural University, Sylhet-3100, Bangladesh.

Md. Rizwanul Islam Tuhin
Faculty of Veterinary and Animal Science, Hajee Mohammad Danesh Science and Technology University, Dinajpur, Bangladesh.

Md. Shafiqul Islam
Department of Pharmacology, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

Abstract/ Executive Summary:

This study was designed with the aim to determine the impact of residual antibiotics on poultry muscle, kidney, spleen, fat & liver and the effect of discriminate and indiscriminate use of antibiotics on hematological constituents in broiler chickens. The chickens were properly reared and on the day of 16, chicks were randomly divided into three groups namely control group (group A), discriminate antibiotic group (group B), indiscriminate antibiotic group (group c). Each group consists of six chicks. In the case of a discriminate group, the chicks were treated with amoxicillin antibiotic followed by maintaining a withdrawal period one week before the sacrifice of the birds. On the other hand, an indiscriminate group of chicks was continued without maintaining any withdrawal period and birds were sacrificed for sampling. TLC method was used for screening detection of amoxicillin residues in poultry tissues and hematological parameters were investigated with appropriate methods such as hemoglobin determination, packed cell volume, and total erythrocyte count. In the case of the control group (group A): No positive sample was found. In the case of the discriminate group (group B): the residues of amoxicillin were 100% in livers, 66.67% in kidneys, 0% in thigh muscles, and 0% in breast muscles, 33.34% in fat, and 33.34% in the spleen. In the case of the indiscriminate group (group c): the residues of amoxicillin were 100% in livers, 100% in kidneys, 16.67% in thigh muscles, and 16.67% in breast muscles, 50% in fat, and 50% in the spleen. Among the poultry tissues, the liver and kidney had the highest level of antibiotic residues in comparison to other samples. It was observed that indiscriminate use of antibiotics represents the antibiotic residues in different edible tissues. Evidence suggests that more judicious use of antimicrobials in food animals will reduce the selection of resistant bacteria and help to preserve these valuable drugs for both human and veterinary medicine. National authorities should adopt a proactive approach that promotes programs aimed at reducing the need for antimicrobials in food animals and ensuring their prudent use. Further investigation is required for the quantitative determination of antibiotic residues in a food product.

Keywords: Antibiotic residues, Amoxicillin, Broiler, Thin layer chromatography

Location: Department of Pharmacology, Bangladesh Agricultural University (BAU), Mymensingh.

Start Date:

End Date:

Program Area: Animal Health and Management

Commodity: Broiler

Objectives:

To detect amoxicillin antibiotic residues in broiler liver, kidney, spleen, fat, breast muscle and thigh muscle by thin-layer chromatography (TLC) test, to assess the impact of antibiotics used as growth promoters on the haematological parameters of broiler chickens and to know the withdrawal period of the drug.

Methodology:

The experiment was conducted in the Department of Pharmacology, Bangladesh Agricultural University (BAU), Mymensingh. The birds were reared in isolated poultry shed, Department of Pharmacology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh. First, the shed was thoroughly washed by sweeping and washing with tap water. The room was disinfected with a phenolic disinfectant (phenyl) and allowed to dry, leaving the room, unused with the electric fan and the bulb switched on. Eighteen apparently healthy day-old broiler chicks were purchased from CP Hatchery Ltd. The chicks were already vaccinated with Marek’s disease vaccine in the hatchery. All chicks were brooded in the same brooder up to 7 days of age under strict management. Vitamin preparation & Glucose was administered to the birds daily mixing with water. BCRDV (Newcastle L-36) & Gumboro vaccine was given to all chicks in the eye (one drop) at 7 and 12 days respectively. Broiler starter mash and broiler grower pellet were obtained from Quality Feeds Ltd. The birds were given ad libitum feed twice daily. Clean & cold water was provided for 24 hours. Amoxicillin BP 30% was obtained as a solution from Navana pharmaceuticals Limited. Chicks were randomly divided on the 16 days of age into three groups. Group A (6 birds) was kept as an untreated control group that received non-medicated water. Group B (discriminate group contain 6 birds) were administered with Amoxicillin at recommended therapeutic dose@ 1gm/5L through drinking water for seven consecutive days from 16^th to 22^th days of age. Group C (indiscriminate group contain 6 birds) were administered with Amoxicillin at recommended therapeutic dose@1gm/5L, through drinking water for fourteen consecutive days from 16th to 29th days of age. Birds received their freshly prepared daily medication in the morning hour of each day. The concentration of Amoxicillin 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. Drinking water was free from pesticides, heavy metals, and coliform count. Drinking water medication was used because oral administration of drugs is the most practical way for treating birds being followed in Bangladeshi field conditions. The weights of every bird were taken individually in the morning and the results were recorded. On day 30^th, before sacrifice; all the birds of three groups (Group- A, B, and C) were weighed individually and the results were recorded. After the slaughtering of the birds, the breast muscle, thigh muscle, kidney, and liver were collected for the estimation of antibiotic residue. All samples were marked separately and preserved at -20º C in polythene bags for their extraction and analysis. Blood samples were also collected and preserved for hematological analysis. For extraction of antibiotic residue from samples thin Layer Chromatography was performed. Samples were blended with a food processor properly for three to five minutes. These mashed/blended samples were taken into properly cleaned and sterilized Petri dishes with proper care as well as covering. From this 4 g of aliquot sample was taken into the beaker with the help of electric balance and spatula. Then homogenization was done with the addition of 10 ml phosphate buffer (pH 6.5). 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 the mixed samples were taken into properly cleaned and sterilized test tubes for centrifugation. Then centrifugation was done at 7000 rpm for 15 minutes with the help of an automatically time-regulated centrifuge machine. 2 ml supernatant was mixed 100 µL of formaldehyde for 45 minutes at 100º C water bath. The supernatant was extracted with an equal volume of diethyl ether and mixing was done properly in order to perform defamation. Then the mixture was kept for 10 minutes to become a separate layer, an upper oily layer, and a bottom layer. Then by using a cleaned and sterilized separating funnel, these mixtures were separated from each other and upper oily was discarded and only the bottom layer was collected. This extraction of supernatant was repeated twice with diethyl ether. Then the extracts were evaporated until dry. The dried sample was reconstituted within 2ml of mobile phase methanol acetone. Then, extracts were collected into the screw-capped vial with proper care and kept in a refrigerator for further advanced analysis. The total procedure was performed as the reference cited by. Amoxicillin BP 30% was used as a reference antibiotic.

Source of Information: International Journal of Natural and Social Sciences, 2020, 7(3): 51-58

Article Link (Online Journal): DOI: 10.5281/zenodo.4063937

Funding Source:

1.

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Indiscriminate use of amoxicillin antibiotic in poultry revealed detectable levels of antibiotic residues in liver, kidney, breast muscle, thigh muscle, fat, and spleen which is dangerous for human consumption. The liver and kidney contained the highest percentage of antibiotic residues. In the study, the level of blood parameters in the control and experimental groups of broiler chickens did not deviate from indices of the control group chickens. Once antibiotics are administered to animal bodies, antibiotic residues are present in high or low concentrations in their products. However, it mainly depends on the duration of the administration of antibiotics. After the administration of antibiotic, concentrations of their residues gradually reduces in meat and other meat products and mostly after 4-5 days of administration secretion of residues are negligible. The antibiotics were identified on the basis of their retention times (RTs) as compared to the standards and by adding known quantities of the standards to the sample and re-chromatography which showed complete overlap of analytic peaks. Hence, the withdrawal time of different drugs should be strictly followed and during this period meat and other animal products like liver, spleen, fat, and kidney should not be used for human consumption. The use of antibiotics as growth promoters should be strictly prohibited and whenever they are used for therapeutic purposes must be used in proper doses and for the proper time. Thus, by observing proper scientific guidelines and precautions we can minimize the harmful effects of antibiotic residues.

Knowledge Management: Journal