Antibiotics Resistance Pattern of Coliform Bacteria Isolated from Slaughterhouse Wastewater in Jega Town, Kebbi State, Nigeria

Wastewater is an essential reservoir of pathogenic bacteria, which include resistant strains. This study determined the antibiotics resistance pattern of Coliform bacteria isolated from slaughterhouse wastewater in Juga local government. Five different samples of wastewater from different locations of a slaughterhouse were collected using standard sample collection techniques. Coliform bacteria were isolated using the standard microbiological method. The total bacterial count was determined using plate count agar. Suspected coliforms were identified based on morphological and biochemical tests. Antimicrobial susceptibility testing was carried out using the standard disc diffusion method. The highest bacterial count (8.4x102 CFU/ml) was observed from the wastewater collection point, and the lowest bacterial (1.2x102 CFU/ml) count from the slaughtering point. Three Gram-negative bacteria, Escherichia coli, Pseudomonas aeruginosa, and Enterobacter aerogenes, were identified. Escherichia coli was the most frequently isolated in slaughterhouse wastewater 39 (45.8%). Among the antibiotics tested against isolated bacteria, Septrin was the most resistant antibiotics recorded against E. coli and E. aerogenes with 84.61% and 88.89% resistant, respectively. Pseudomonas aeruginosa was found to show higher resistance to Chloramphenicol and Septrin with 84.21% resistance each. The occurrence of antibioticresistant bacteria from slaughterhouse wastewater showed the risks associated with antimicrobial drug resistance transferred from foodproducing animals to humans. Management concerns, such as local government health officers and community development officers, should increase the sensitization of slaughterhouse workers by organizing conferences or conducting radio talk to educates these slaughterhouse workers on the treatment of slaughterhouse wastewater and health risk associated with antimicrobial-drug resistance transferred from animals to humans.


INTRODUCTION
The wastewater is an essential reservoir of bacterial pathogens, including resistant isolates (Egesi et al., 2019).
Wastewater from the slaughterhouse, if not carefully treated or disposed of, can be good sources of antibiotics resistant bacteria (Manyi-Loh et al., 2018), mainly where slaughterhouse is located near the vegetable farms or community sources of drinking water (Egesi et al., 2019).
Activities of meat processing from slaughterhouses in Nigeria are mostly handled by people who are unaware of the slaughterhouse and environmental safety policies (Johnson & Etokidem, 2019), especially regarding possible microbial contaminations during meat processing (Egesi et al., 2019). Wastewater from

Abstract
Wastewater is an essential reservoir of pathogenic bacteria, which include resistant strains. This study determined the antibiotics resistance pattern of Coliform bacteria isolated from slaughterhouse wastewater in Juga local government. Five different samples of wastewater from different locations of a slaughterhouse were collected using standard sample collection techniques. Coliform bacteria were isolated using the standard microbiological method. The total bacterial count was determined using plate count agar. Suspected coliforms were identified based on morphological and biochemical tests. Antimicrobial susceptibility testing was carried out using the standard disc diffusion method. The highest bacterial count (8.4x10 2 CFU/ml) was observed from the wastewater collection point, and the lowest bacterial (1.2x10 2 CFU/ml) count from the slaughtering point. Three Gram-negative bacteria, Escherichia coli, Pseudomonas aeruginosa, and Enterobacter aerogenes, were identified. Escherichia coli was the most frequently isolated in slaughterhouse wastewater 39 (45.8%). Among the antibiotics tested against isolated bacteria, Septrin was the most resistant antibiotics recorded against E. coli and E. aerogenes with 84.61% and 88.89% resistant, respectively. Pseudomonas aeruginosa was found to show higher resistance to Chloramphenicol and Septrin with 84.21% resistance each. The occurrence of antibioticresistant bacteria from slaughterhouse wastewater showed the risks associated with antimicrobial drug resistance transferred from foodproducing animals to humans. Management concerns, such as local government health officers and community development officers, should increase the sensitization of slaughterhouse workers by organizing conferences or conducting radio talk to educates these slaughterhouse workers on the treatment of slaughterhouse wastewater and health risk associated with antimicrobial-drug resistance transferred from animals to humans.
In Nigeria, monitoring environmental sanitation and waste management were established and implemented (Egwuenu et al., 2018). However, the agency lacks monitoring to assure the proper practice of these policies by some local community, which leads to the spread of these commonly resistant bacteria within the community. Research on resistant bacteria is fundamental as this will provide some preliminary information about the existence of these bacteria within the local community (Aslam et al., 2018). However, literature has shown a lack of comprehensive data regarding resistance to antimicrobial agents, especially in African continents, including Nigeria (Egwuenu et al., 2018). Therefore, this study was designed to determine the antibiotics resistance pattern of coliform bacteria isolated from slaughterhouse wastewater.

Research sites
This study was carried out in the Jega local government area of Kebbi State (Figure 1). The local government is located at the latitude and longitude 12.

Isolation and identification of coliform bacteria
Coliform bacteria were isolated using the surface spread plate method described by Cheesbrough (2006) and Bashir et al. (2018). Total coliforms and bacterial counts of the collected wastewater samples were determined using Eosin Methylene Blue (EMB) agar and Plate count agar.
Serial dilutions of the water samples were made aseptically by transferring 1 mL of the wastewater sample into 9 mL of sterile distilled water and diluting that range from 10 -1 to 10 -6 . After that, an aliquot of 0.1 mL from 10 2 dilutions was plated on Plate count agar (Himedia) in duplicates then spread aseptically using a sterile spreader (Egesi et al., 2019). Plates were incubated at 37°C for 24 hours and colonies were counted and expressed as colonies forming units per milliliter (CFU/mL).  (Holt et al., 1994).

Antimicrobial susceptibility test
Antibiotic susceptibility patterns of the pure colony of E.

RESULTS AND DISCUSSION
A study on antimicrobial-resistant is critical now a day mainly from the African continent, where comprehensive data to show the magnitude of resistance are lacking (Egwuenu et al., 2018). Literature has shown that wastewater from slaughterhouse can be served as a source or reservoir of resistant bacterial pathogens spread within the community if not well treated or disposed of (Egesi et al., 2019). This resistance varies from different locality due to some factors such as the educational background of animal rearers, use of antibiotics in animal feedstuffs, among others (Egwuenu et al., 2018). This hinders the effort to control antimicrobial-resistant humans because resistant bacteria of animal origin are more dangerous than human origin (Hiroi et al., 2012).
This research determined the antibiotics resistance pattern of coliform bacteria isolated from slaughterhouse wastewater in Jega local government area Kebbi State, Nigeria.
Out of five samples of wastewater collected from different slaughterhouse points, the result showed that the wastewater collection point had the highest bacterial coliform counts with a mean and standard deviation of (8.73 ± 0.35x10 2 CFU/mL) while the Slaughtering point had the lowest coliform bacterial counts with mean and standard deviation of (1.27 ± 0.31x10 2 CFU/mL) as presented in Table I   The suspected coliform bacteria were identified using a standard biochemical test, as shown in Table II. Among the coliform bacteria isolated from the different slaughterhouse sampling points, E. coli is the most abundant bacteria isolated with 39 points (45.8%), as presented in Table III. The presence of E. coli from slaughterhouse wastewater is an indication of fecal contamination, and therefore such water needs to be given special treatment to arrest the spread of this pathogenic organism within the environment. Disease caused by E. coli of animal origin is more difficult to treat, especially the resistant isolate due to the frequent use of antibiotics in animal feeds (Hiroi et al., 2012). This is in line aerogenes. The presence of P. aeruginosa in slaughterhouse wastewater was also reported by previous studies (Igbinosa et al., 2012;de Oliveira et al., 2013;Ayogu et al., 2018). This confirmed the statement by Yayan et al. (2015) that P. aeruginosa is currently one of the dangerous nosocomial organisms that can be transmitted in many ways, including food products such as meat.
The percentage (22.4%) of P. aeruginosa reported in this study is lower than the 37.6% reported by Rabiu and Falodun (2017) from wastewater of a slaughterhouse in Ibadan, Nigeria.

(100)
The result of antibiotics resistant pattern of the coliform bacteria isolated (Figure 2) indicates that E. coli showed higher resistance to Septrin and Augmentin among the antibiotics tested with (84.61%) and (89.74%) resistant, respectively. The higher resistance of E. coli recorded in this research is worrying as recent literature showed that many of the antimicrobial-resistant strains of E. coli that cause human urinary tract and bloodstream infections appear to have arisen from contaminated meat products (Egwuenu et al., 2018). The finding of this research is in line with the report of Atuanya et al. (2018) and Savin et al. (2020), who reported the resistance of E. coli isolated from slaughterhouse Effluents to Septrin and Augmentin.
Similarly, Jega et al. (2018)    This bacterium's resistance to this drug is alarming as this bacterium was reported to be one of the dangerous nosocomial microorganisms that can be transmitted in many ways, including food products such as meat (Yayan et al., 2015). Infection caused by P. aeruginosa, exceptionally resistant isolates seems challenging to treat due to its resistance to a variety of antibiotics, and patients can die from pneumonia caused by Pseudomonas spp. (Tran et al., 2014). The increasing resistance of P. aeruginosa

CONCLUSION
The occurrence of antimicrobial-resistant pathogens such as E. coli, E. aerogenes, and P. aeruginosa from this slaughterhouse points showed the risks associated with antimicrobial drug resistance transferred from foodproducing animals to humans. It is recommended that the management concern, such as local government health officers, and community development officers should increase on the sensitization of slaughterhouse workers by organizing conferences or conducting radio talk to educates these slaughterhouse workers on the treatment of slaughterhouse wastewater and health risk associated with antimicrobial drug resistance transferred from food-producing animals to humans.

ACKNOWLEDGMENT
The author thanks all staff of the Microbiology laboratory of Kebbi State University of Science and Technology, Aliero, for their kind support during this study.