The study was conducted at different area of Nalitabari paurosova (Figure 1) in

Sherpur district of Bangladesh during the periods of February 2015 to December 2018. Recently, huge number of poultry firm were established in this area that was an important reason for analyzing the poultry meat and all the samples were collected from poultry farms located in this region.

Fifteen samples were aseptically collected from study area. The slaughtered broilers at first immersed in a special tank containing hot water for some time. The immersed birds were de-feathered traditionally by hand plucking and subsequently evisceration was done using special tricks or techniques. The muscle of breast region were cut and put into a sterilized container. During transportation the sterile containers were kept cool in iceboxes containing fragments of ice.

The anthropometric data were collected based on literature review of similar existing study [36] . In this study, randomly 400 respondents were selected by semi structured questionnaire based cross-sectional population selection system from study area. Among total respondents, 200 were females and 200 males. Age of the subjects under this study was determined by interrogation and confirmed with birth certificate or the health card. Measurements of weight were obtained by digital machine at three times and the average was calculated with minimal clothes and bare footed. The height was measured using a measuring tape without shoes and the average was calculated and recorded with standard error. Body Mass Index (BMI) was computed using the following standard equation: BMI = Weight (kg)/height (m²) [37] . Based on the interrelationships of height, weight, age, height for age, weight for age and height for weight have been calculated to determine their nutritional status [38] . Nutritional status (such as: thinness, malnourished, obesity, normal weight and overweight etc.) was evaluated following the recently published international BMI cut off points [39] .

Each of the raw meat samples was macerated in a mechanical blender using a sterile diluent as per recommendation of International Organization for Standardization (ISO). Ten grams of the breast meat sample was taken aseptically with a sterile forceps and transferred into sterile containers containing 90 ml of 0.1% peptone water. A homogenized suspension was made in a sterile blender. Thus 1:10 dilution of the samples was obtained. Later on, different serial dilutions ranging from 10⁻² to 10⁻⁹ were prepared according to the standard method [40] .

For the determination of TVC and TCC, 100 µl of each ten-fold dilution were transferred and spread on Plate Count Agar (PCA) and MacConkey agar using a fresh pipette for each dilution. The diluted samples were spread as quickly as possible on the surface of the plate with a sterile glass spreader. The plates were then kept in an incubator at 37˚C for 18 hours. The average number of colonies in a particular dilution was multiplied by the dilution factor to obtain the total viable count. The TVC and TCC were calculated according to ISO [40] . The results of the total bacterial count were expressed as the number of organism or colony forming units per gram (CFU/gm) of meat sample [41] .

For the identification of pathogenic bacteria, 100 µl of each sample were transferred into Thiosulfate Citrate Bile Salts Sucrose Agar (TCBS) media and Shigella Salmonella Agar (SS) media with ten-fold dilution. The diluted samples were spread as quickly on the surface of the plate with a sterile glass spreader and incubated at 37˚C for overnight. The presence of pathogenic bacteria were observed and counted.

The data on TVC and TCC obtained from the bacteriological examination of meat samples of the poultry carcass collected from different area of Nalitabari. Data were analyzed with Microsoft Office Excel-2013 [41] .

The cultural examination of chicken breast meat samples for bacteriological analysis was done according to the standard method by International Commission on Microbiological Specifications for Foods (ICMSF) [42] . The examination followed detail study of colony characteristics including the morphological and biochemical properties. In order to find out different types of microorganisms in chicken breast meat samples, different kinds of bacterial colonies were isolated in pure culture from the Plate Count Agar (PCA), MacConkey agar, SS agar and TCBS agar and subsequently identified according to the methods described by Krieg et al. [43] . Gram staining and biochemical reaction were performed for further confirmation of presumptively identified bacteria according to Bergey’s Manual Determinative Bacteriology [44] . Among these 15 samples 12 kinds of biochemical test such as Kligler Iron Agar (KIA), Motility-Indole-Urease (MIU), Citrate, Voges Proskauer (VP), Oxidase, Catalase, Mannitol, Starch, Methyl Red (MR), Glucose, Lactose, Eosin Methylene Blue (EMB) were performed.

The antibiotic susceptibility of the Escherichia coli and Salmonella spp. isolates was determined according to the standard disc-diffusion method [45] . Well isolated single colony obtained from overnight grown cultures, were used for making young culture for the test. The commercially available antibiotic discs (Oxoid, UK) used in this study were: Amoxicillin (30μg), Ampicillin (10μg), Tetracycline (30μg), Nalidixic Acid (30μg), Ciprofloxacin (5μg), Norfloxacin (10μg), Erythromycin (15μg) and Gentamicin (10μg).

Healthy and disease free plant leaves of Azadirachta indica were collected from nearby area. The freshly collected leaves were washed with tap water, 70% ethanol and then distilled water to sterilize and remove the external impurities. These were further slice into small pieces and shade dried for two weeks and then blended into powder using mortar. The powdered leaves of A. indica were separately extracted with different solvents (30%, 40% of ethanol and methanol) by using Whatman No. 1 filter paper. The crude extracts were obtained by concentrating the mixture solution using a rotary evaporator and used for further tests.

Young culture of bacterial aliquots of the test organisms in 4 ml sterile Mueller Hinton Broth (MHB) were made from well isolated single colony obtained from 24 h growth cultures. Each aliquots (10 μl) containing approximately 5 × 10⁴ bacterial cells or colony forming units was transferred into Mueller-Hinton Agar (MHA) plates. The disc containing plant extract were diffused into the plate where, the extract allowed to stands for an hour for reaction to take place between the extracts and the bacterial organisms. The plate were then inoculated on separate and incubated at 37˚C for 18 hours.

Three kinds of bacteria (E. coli, Salmonella spp. and Vibrio spp.) were isolated by observing distinct morphological characteristics on selective media and further confirmed with standard Biochemical test (Table 2).

K = Alkaline, A = Acid, G = Gas, AG = Acid and Gas, + = Presence, − = Absence.

Resistance against commercially available commonly used antibiotics has been observed in bacteria present in broiler since the introduction of these antimicrobial agents in poultry. The rise in antibiotic resistance has been reported in the past two decades in many countries including Bangladesh [66] . Based on the susceptibility to antibiotics, the bacteria were categorized into three groups—sensitive, intermediate, and resistance. Total 20 isolates (10 E. coli and 10 Salmonella spp.) were randomly selected for antibiotic susceptibility test.

In this study, E. coli showed highly resistance to Ampicillin (80%) and Amoxicillin (70%). Where, 40% were resistance to Nalidixic acid and Erythromycin. About 90% E. coli showed sensitivity to Ciprofloxacin, 80% to Gentamicin, 60% to Tetracycline and 50% to Norfloxacin (Figure 2). The results strengthen the earlier observations of some study where it was found that the E. coli isolated from broiler were sensitive to Azithromycin, Ciprofloxacin, Norfloxacin and Gentamicin and resistant to Amoxicillin and Erythromycin. The possessions of such factors by the E. coli isolates signify the fact that the organisms might have gained the resistance property due to the unsystematic use of antibiotics [67] . Salmonella spp. showed 100% resistance to Ampicillin, 90% resistance to Amoxicillin and highly sensitive to Ciprofloxacin (80%). About 50% were resistance to Nalidixic Acid and Norfloxacin whereas sensitive to Erythromycin (60%) and Gentamicin (50%) (Figure 3). Some others earlier study also revealed that

Salmonella spp. were sensitive to Ciprofloxacin, Gentamicin and Azithromycin [68] [69] and resistant to Erythromycin and Amoxicillin [70] [71] . Potential drug resistant pathogens in normal broilers may be a serious public health concern.

The zone of inhibition of negative (30% methanol, 40% methanol, 30% ethanol, 40% ethanol, DMSO) control of plant extract and positive control (Ciprofloxacin) with and their comparison is presented in (Table 3). The above result specified the antimicrobial activity of extract against Escherichia coli and salmonella spp. depending on the nature of the active ingredients present in the extracts and their capacity of diffusion into agar medium. Antibacterial activities of methanolic and ethanolic plant extract (both 30% and 40%) were significant against E. coli. The average zone of inhibition of methanolic and ethanolic extract were 13 and 14.33 respectively, where 40% ethanolic plant extract was more potent than the 30% (Figure 4). No zone of inhibition was formed against isolated Salmonella spp.

N/A = No Activity.

A previous study found the average zone of inhibition of methanolic extract of Azadirachta indica against Escherichia coli was 7.5 mm [72] . The zone of inhibition against E. coli in both methanolic and ethanolic extract was not found by another research [73] . But Zone of inhibition of methanolic extract of same plants against Salmonella spp. was reported as 10 - 20 mm [74] . Another study evaluated the zone of inhibition with different concentrations of methanolic plant extract were 20 mm, 22 mm, 24 mm and 21 mm in diameter against E. coli, where 18 mm, 22 mm, 20 mm and 21 mm in diameter against Salmonella spp. [75] .

The total respondents were divided into five groups based on age range—5 to 15, 16 to 30, 31 to 40, 40 to 50 and above 50 years old (Table 4). Each group had 40 male and 40 female respondents (Table 4). Total 300 (75%) people eat broiler meat among the total respondents, where 150 (50%) were females and 150 (50%) were males. The educational status of the respondent showed that, about 8% were illiterate, 10% completed primary level, 21% passed up to secondary level, 32% were higher secondary level and 35% were in graduation level (Table 5).

On the basis of family income, the respondents were divided into five socio-economic groups, like poor (15.75%), lower middle class (23.5%), middle class (49.25%), upper middle class (6.75%) and higher class (4.75%) (Table 6). Most of the female respondents in this study were domestic worker (34.5%). The major source of family income among these respondents were day laborer (18%) and businessman (18%), rest of them were private service holder (14.75%) and government service holder (14.75%) (Figure 5).

In this study, it was found that consumption of broiler meat largely varies with the socio-economic status of respondents (Table 7). Depending on economic status, most of the poor people (N = 31, 49.20%) ate broiler meat at least one day in a week where very few people of upper middle class (N = 9, 21.95%) and higher class (N = 2, 4.88%) ate occasionally, and majority of last two groups never eat broiler meat (N = 12, 44.44%) and (N = 17, 89.47% respectively).Large number of broiler meat consumer were middle classes and lower middle classes

where most of the mate broiler meat frequently (N = 85, 73.91%) and At least 2 - 4 days in a week (N = 44, 61.97%). Very few people of middle and lower middle classes ate broiler meat at least 5 days in a week (Figure 6).

BMI is an important consideration for evaluating the health condition of particular people. Various BMI had been observed among the responders who ate broiler meat or not. By considering various BMI group, it was found that, overweight (male 24.67%, female 22.67%) and obesity (male 32%, female 8%) of the responders who ate broiler meat were high in comparison with overweight (male 10%, female 16%) and obesity (male 8%, female 6%) of responders who never ate broiler meat (Figure 7).

Some common diseases such as food allergy, overweight, obesity, high blood pressure, and diabetes were considered to determine the possible correlation with consumption of broiler meat. Food allergy is allergic reaction of food includes fish, shellfish, peanuts, tree nuts, walnuts, etc., but not directly related to broiler meat. In this study, it was found that the occurrence of food allergy relatively high among the people who never ate broiler (male 32%, female 12%) compare to the people who ate broiler meat (male 23.3%, female 9.3%), whereas, in case of other diseases, it was vice versa. Among the broiler meat consumer, overweight, obesity and high blood pressure were found in elevated rate than any other diseases both in male and female. Male who eat broiler meat had high blood pressure, high cholesterol level, digestive disorders, constipation and diabetes in alarming percentage with 35.3%, 22.7%, 18.7%, 24% and 7.3% respectively whereas male who never eat broiler meat had this diseases in percentage with 16%, 10%, 14%, 6% and 4% in some respect. Surprisingly, it is a matter of great concern that, 3.3% male who eat broiler meat found having benign fat deposition like tiny tumor but this disease did not found in male and female who never eat broiler meat (Figure 8).