The term ‘poultry’ used in agriculture generally refers to all domesticated birds kept for egg laying, meat production, or their feathers. Poultry comes from the French word ‘poul’, which was derived from the Latin word 'pulus' meaning small animals. Poultry meat and its products are among the most widely consumed foods in the world. Chicken meat is delicious, nutritious, and a good source of protein. It is characterized by a good flavor and is easily digested. Poultry farms have appeared to be a successful and widely spread business industry in Nigeria, which often remains contaminated with various hazardous microorganisms when standard hygiene practices are compromised. Poultry remains the largest domestic animal stock in the world in terms of the number of animals. In Nigeria, more than three million people are employed directly in the poultry sector, which provides the largest supply of meat and eggs, so as to meet the major protein sources for the entire population of the country (Nandi et al., 2013). Poultry is a fast-growing source of meat in the world today, representing a quarter of all the meat produced [1]. Meat and poultry products are some of the sources for transmitting food-borne pathogens to humans, with 40% of the clinical cases attributed to the consumption of chicken, eggs, and other poultry products [2-4]. The incidence of food-borne diseases in humans has increased considerably worldwide in the last few years. Poultry products have been repeatedly implicated in food-borne infections, as poultry birds can harbor different food-borne pathogens. Many reports in recent years have shown that Salmonella and Campylobacter spp. are the most common causes of human food-borne bacterial diseases linked to meat and other poultry products like eggs [5]. Food-borne infections and intoxications have been estimated to cause about one billion cases of acute diarrhea annually in children under the age of 5 in Africa, Asia, Latin America, and other developing countries [6]. Poultry feeds are infected during processing through handling, mixing of ingredients, and exposing the raw materials and finished products to atmospheric microorganisms. The most common vehicles for transmission of food-borne salmonellosis are meat, meat products, eggs, and egg products that are contaminated as a direct result of animal infection [3]. The problem of non-typhoid Salmonella in Africa is very serious but has generally been overshadowed by these three epidemic diseases: malaria, human immunodeficiency virus (HIV), and tuberculosis [7]. Prominent bacterial species in the poultry farms include Escherichia coli, Enterococcus, Proteus, Clostridium, Salmonella, Providencia, Aeromonas, and Lactobacillus that have been shown to be of critical importance in tropical countries [8-10] and elsewhere in the world.

The glassware was properly washed, air dried, wrapped with aluminum foil paper, and sterilized in a hot air oven at 180°C for 2 hours.

Sample Collection

A total of twenty-five (25) samples were collected from both the cloaca and oropharyngea of poultry birds from five different poultry farms. Sterile swabs sticks moistened with sterile normal saline were used to take samples from the cloaca and oropharyngea of five randomly selected birds in each poultry farm. After collection, all samples were transported immediately to the laboratory for analysis.

Isolation and Identification of Bacteria

The swab sticks containing the samples from cloaca and oropharyngea were streaked on already prepared nutrient agar plate and incubated at 37 °C for 24 hours. Growth was observed on each plate, and developed colonies were subcultured on MacConkey agar to obtain distinct colonies. The pure bacterial isolates were identified based on their morphology, Gram reaction, and biochemical tests. Distinct colonies were subcultured to obtain pure isolates, which were stored at 40 °C for microscopic characterization and biochemical tests.

Antibiotic Susceptibility Tests

The disc diffusion was used to examine bacterial susceptibility to antimicrobial agents. A total of ten (15) antibiotics containing Streptomycin 10µg, Erythromycin 15µg, Chloramphenicol 30µg, Ciprofloxacin 10µg, Tetracycline 30µg, Amoxil 20µg, Augumentin 30µg, Ofloxacin 10µg, Pefloxacin 10µg, Streptomycin 10µg, Levofloxacin 20µg, Ciprofloxacin 10µg, Chloraphenicol 30µg, Norfloxacin 10µg, and Rifampicin 20µg were used. Each bacterial isolate was adjusted to a 0.5 McFarland standard, and sterile swab sticks were dipped into the adjusted suspensions. The swab was then spread evenly over the entire surface of the already prepared Mueller-Histon agar plates to obtain uniform inoculation. The plates were then allowed to dry for 3-5 minutes. Antibiotics were then placed on the surface of the inoculated plates with sterile forceps. Within 15 minutes of the application of the disc, the plates were inverted and incubated at 37⁰ °C. After 16–18 hours of incubation, the plates were examined, and the diameter of the zones of complete inhibition to the nearest whole millimeter was measured. The zone diameter for individual antimicrobial agents was then translated into susceptible, intermediate, and resistance categories according to the interpretation table of Becton Dickinson Microbiology Company, U.S.

The results from this study show the probable bacterial isolates identified on the basis of cultural characteristics, morphological features, Gram staining reactions, and biochemical characteristics of the bacterial isolates (Table 1).

The bacterial isolates from the cloaca and oropharyngeal swabs include Staphylococus spp, Escherichia coli, Streptococcus sp, Bacillus spp, Salmonella spp and Shigella sp. The percentage occurrence of the bacterial isolates from both cloaca and oropharyngea shows that the bacterial isolates were found predominantly in oropharyngeal swabs in comparison with the cloaca swabs (Tables 2–3).

Antibiotic susceptibility results show the bacterial isolates have multiple resistances to the selected antibiotics used, while few isolates were susceptible to these antibiotics.

(Table 4-5).

Table 1 shows the morphological identification, Gram reaction, and biochemical test results conducted on the isolates and probable organisms from both the cloaca and oropharyngeal. The organisms for cloaca samples were Streptococcus sp.aphylococcus sp., Salmonella sp.cillus sp., and Escherichia coli, while the organisms for oropharyngeal samples were all isolates from Cloaca and Shigella sp.

Table 1: Morphological Characteristics, Gram Reaction and Biochemical Characteristics of Isolates.

Key: CIT= Citrate, CAT= Catalase, MOT= Motility, IND=Indole, UREA= Urease, H₂S = Sulphur utilization, + = Positive,   - = Negative

Table 2 shows the frequency distribution of bacteria isolated from the cloaca of the poultry from the different poultry farms. Streptococcus sp had the highest percentage occurrence (32.2%). Escherichia coli and Staphylococcus sp. had the least percentage occurrence (12.9%).

Table 2: Frequency Distribution of Bacterial Isolates from Cloaca of Poultry Birds.

Table 3 shows the frequency distribution of bacterial isolates from the oropharyngeal of the five farms. Shigellasp had the highest percentage occurrence (25.9%). Escherichia coli had the lowest percentage occurrence (7.4%).

Table 3: Frequency Distribution of Bacterial Isolates from Oropharyngeal of Poultry Birds.

Table 4 shows the antibiotic susceptibility tests of Gram-negative isolates. Shigellasp and Salmonellasp had the highest percentage resistance of (80%) to the antibiotics used. Escherichia coli had the lowest percentage resistance (40%) to the antibiotics used.

Table 4: Antibiotic Susceptibility Test of Gram Negative Isolates.

Key: CPX= Ciprofloxacin, AM=Amoxil, GN= Gentamycin, OFX= Ofloxacin , AU= Augurmentin, CH= Chloramphenicol, S=Streptomycin, PEF = Perfloxacin. SP= , SXT= R= Resistance (0-12mm), S=Sensitive (> 18mm), I= Intermediate (13-17mm).

Table 5 shows the antibiotics. Susceptibility test of gram-positive isolates Streptococcus sp had the highest percentage resistance (70%). Bacillus spp. had the lowest percentage resistance of 30%.

Table 5: Antibiotic Susceptibility Test of Gram Positive Isolates.

Key: CPX= Ciprofloxacin, AMX= Amoxil, NB= Norfloxacin, APX= Ampixcylin, RD= Rifampicin, CH= Chloramphenicol, LEV=Levofloxacin, E=Erythromycin, S = Streptomycin, CN=

R= Resistance (0-12mm), S=Sensitive (> 18mm), I= Intermediate (13-17mm).

The result revealed that a total of six (6) bacteria-Streptococcus sp., Bacillus sp., Escherichia coli, Staphylococcus sp., Salmonella sp and sp.,igella sp.—were isolated from the cloaca and oropharyngeal tissues of poultry birds and identified on the basis of morphological and biochemical characteristics. This is in agreement with the work of Khan [11] who also isolated similar bacteria from poultry farms. The occurrence of these bacterial species in public health concerns may indicate an obvious health hazard in terms of consumption of bacteriologically contaminated poultry meat [12]. Oropharyngeal and cloaca are important route openings in poultry birds for the ingestion and ejection of food and waste. These routes favor the growth of many bacterial species; this could be as a result of the availability of favorable temperatures and the presence of materials the birds feed on [12].

The presence of these bacteria pathogens in poultry birds could be a result of contaminated feed and water taken by the birds; this is consistent with the findings of Okoli [12], who reported that commercial feeds are an important vehicle for the introduction of pathogens into poultry birds. The presence of Salmonella and Escherichia coli isolated from this study suggests fecal as well as environmental contamination. These organisms are well-known pathogens of birds and farmed animals. E. coli, for example, was reportedly implicated in disease conditions such as colibacillosis, which occurs in forms such as enteric and septicaemia colibacillosis, whereas Salmonella is capable of producing acute and chronic infections in all or most types of birds and animals [13]. The water from the feeding operations for poultry is an excellent source of harmful bacteria [13].  Rujuta [14] observed the presence of Salmonella and Escherichia coli in water samples collected from poultry farms, reporting that this could be due to human activities, soil contamination, and, conjointly, the ability of the organism to survive throughout a large selection of habitats. The water provided to chickens could be involved in the spread of these pathogens among poultry and their potential transmission to humans.

The presence of Streptococcus, Staphylococcus, and Bacillus species indicates contamination from feed. Following the report of Uwaezuke and Ogbulie [15], they found these isolates, Staphylococcus Streptococcus, to be the major poultry feed contaminants in Nigeria. These organisms are pathogenic, causing both upper respiratory tract infections like Streptococcal pharyngitis and alimentary tract infections like diarrhea, both in birds and humans, which has been a food-borne illness reported by Dasheet [9]. From this study, the bacteria isolated are harmful to humans. This suggests a lack of sanitation and proper hygiene management. The presence of these pathogens in chickens makes them carriers, and then the bacteria are transferred to humans through the consumption of poultry meat. Various gastrointestinal diseases are caused in humans by the consumption of contaminated poultry meat.

Most of the isolates in this study exhibited multiple resistances to the antibiotics used. Similar findings on multiple resistance were observed by Rahman, who reported that the significant increase in the incidence of resistance against antibiotics isolated from chickens is probably due to the increased use of antibiotics as feed additives for growth promotion and prevention of diseases.

This study shows the presence of potentially pathogenic bacteria, which are usually of public health significance, and that the development of resistance to antibiotics can be attributed to long-term usage of antibiotics as therapeutics and growth promoters. Hence, the appropriate use of antibiotics in farm animals needs to be addressed.

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