Skip to main content
Download PDF

Role of migratory birds as a risk factor for the transmission of multidrug resistant Salmonella enterica and Escherichia coli to broiler poultry farms and its surrounding environment

BMC Research Notes volume 17, Article number: 314 (2024) Cite this article

Abstract

Multidrug resistance (MDR) considered as global crisis facing poultry industry. Migratory birds play very important role in the dissemination of antimicrobial resistant pathogen during their fly way specially to poultry farms. Therefore, 750 samples from migratory birds and 300 samples from broiler chicken farms and its environment were collected during the winter seasons of five years (2019 to 2023). The samples were subjected to the isolation of Salmonella enterica and Escherichia coli with the detection of antimicrobial resistance (phenotypic and genotypic) with insight to the genetic similarity between the isolates from migratory birds and broiler chickens’ farms. Different members of Enterobacteriaceae were isolated; Salmonella enterica, Escherichia coli, Citrobacter, Enterobacter, Klebsiella, Proteus, Providencia, Serratia, Hafnia. 298 (28.4%) of S. enterica strains belonging to 27 serovars. S. Typhimurium, S. Kentucky, S. Enteritidis and S. Shangani were the common 4 serotypes between migratory birds and farms. Meanwhile, we found 489 (46.6%) isolates of E. coli belonging to 24 serogroups and O91, O128, O26, O125, O55, O103 and O159 were the common 7serogroups between migratory birds and farms samples. The majority of Salmonella (91.6%; 274 out of 298) and E. coli (92%; 450 out of 489) were MDR. The MDRI range of Salmonella and E. coli was 0.08- 1.The genetic similarity between the isolates of migratory birds and broiler chicken farms were detected by ERICPCR and hot map. This study suggests the continuous applications of surveillance programs for migratory birds and biosecurity measures in poultry farms.

Peer Review reports

Introduction

In the Middle East, Northern Africa, and the Mediterranean basin, Egypt considered as one of the higher population densities [1]. Bird migration is one of nature’s great mysteries and spectacles, and Egypt and the Middle East are in the epicenter of one of the world’s major migration pathways [2]. Migratory birds have the ability to spread Salmonella to human via shared environment, direct contact and fecal shedding [3], as well as to domestic poultry [4], resulting in both animal and human illnesses as well as significant economic losses to the poultry industry [5]. During the bird’s migration, significant impacts on the ecology and dissemination of potentially pathogenic antimicrobial resistant bacteria, including ESBL-producing E. coli [6] and MDR non-typhoidal Salmonella [7] have been arise.

Salmonellosis is one of the most important problems facing poultry industry [8,9,10] which results in dehydration, diarrhea, body weight loss, arthritis, pneumonia and omphalitis in the infected birds [11]. This clinical symptom is provoked by Salmonella Gallinarum and Pullorum and the birds can harbor asymptomatic Nontyphoidal Salmonella so, they are considered as silent reservoirs [12].

Also, the majority of E. coli produces beneficial effects such as protecting against other harmful bacteria, but when it acquires genetic material from other organisms, it converts into pathogenic [13]. The main cause of colibacillosis in poultry is avian pathogenic E. coli [14], it results in mortality rates of up to 30% [11], which is a significant issue for the Egyptian poultry sector [15].

Antimicrobial resistance is a serious health concern worldwide [16]. Antimicrobial resistance may be attributed to frequently use of antimicrobial drugs for therapy or as growth promoters, and poultry remains a significant source of zoonotic MDR bacteria [17]. This study was aimed to determine the role of migratory birds as a risk factor in the transmission of Salmonella and E. coli to broiler farms over a period of 5 years in three Egyptian Governorates in addition to conducting examinations on the antimicrobial susceptibility and genetic diversity of Salmonella and E. coli isolates.

Materials and methods

Samples collection

This study targeted 3 Egyptian Governorates in the northern delta of Egypt (Dakahlia, Damietta and Port Said). It began from the year 2019 and continued for 4 consecutive years until 2023. The sampling was conducted from October to February during each year. Thousand and fifty samples were collected including 750 sample from migratory birds (150 birds each year including 50 birds from each Governorate) that was found near to broiler farms and that sold in live bird markets in addition to 300 samples from broiler chicken and its surrounding environment. The collected migratory birds marked and photographed in order to find out the English and scientific names. The samples were included cloacal swabs from migratory birds in addition to swabs from cages and surrounding environment in the bird markets which were collected by using sterile cotton swabs pre-moisten with Buffered Peptone Water (2 ml) according to the methods described by ISO 6579-1 [18].

On the level of broiler chicken farms, the samples were collected from 300 diseased broiler chicken aged 25–40 days (60 farms each year; 20 farms from each Governorate). Clinical examinations and postmortem (PM) lesions were recorded in the investigated farms. From each farm, 10 diseased chickens were selected, sacrificed and subjected to PM examinations under septic conditions whereas samples from internal organs (liver, spleen, cecum, heart and lung) were aseptically collected by using a scalpel to cut a piece of tissue (approximately 4 cm3) and by sterile forceps the tissue was placed into a sterile container. The tissues were pooled together and mixed with 20 mL of BPW by stomaching for 30 s for each bird individually [19].

Bacteriological examinations

The collected samples were subjected the isolation and identification of Salmonella and E. coli. Salmonella was isolated on Xylose Lysine desoxycholate (XLD) agar plates and biochemically identified according to ISO 6579 [18]. The confirmed Salmonella isolates were subjected to serological identification according to Kauffman – White scheme [20] to determine Somatic (O) and flagellar (H) antigens using Salmonella antiserum (DENKA SEIKEN Co., Japan).

E. coli was isolated on Eosin Methylene blue agar plates and biochemically identified according to Lee and Nolan [21]. The confirmed E. coli biochemically was serologically identified according to Quinnet al. [22]. by using rapid diagnostic E. coli antisera sets (DENKA SEIKEN Co., Japan).

Antimicrobial susceptibility testing

The in vitro antimicrobial susceptibility of the confirmed Salmonella and E. coli was detected using the disc diffusion method on Mueller–Hinton agar (Oxoid, UK) according to the guidelines stipulated by Clinical Laboratory Standards [23] whereas the tested antimicrobial discs were categorized into sensitive, intermediate and resistant. The basis of the antimicrobial agent selection was owed to their frequent use in poultry farms in addition to their importance for human and veterinary fields. Twelve discs belonging to 8 classes were selected; quinolones (nalidixic acid; NA − 30 µg, ciprofloxacin; CP- 5 µg and levofloxacin; L- 5 µg), tetracycline (tetracycline; - T 30 µg), penicillin (ampicillin; AM- 10 µg), sulfonamides (sulfamethoxazol; SXT- 25 µg), lincosamides (clindamycin; CL- 10 µg), aminoglycosides (gentamicin; G- 10 µg, amikacin; AK- 30 µg and kanamycin; K- 30 µg), cephalosporin (cefotaxime; CF- 30 µg) and macrolides (erythromycin; E- 15 µg).

Salmonella Typhimurium ATCC14028 and E. coli ATCC 25,922 were used as control strains. MDR strains of the isolated Salmonella and E. coli were represented to exhibit resistance to three or more different antimicrobial classes [24]. Additionally, MARI was calculated using the formula (Number of antimicrobials showed resistance in each isolate/ Total number of the tested antimicrobial agents).

Molecular detection of antimicrobial resistant genes

Based on the results of the antimicrobial susceptibility testing, conventional polymerase chain reaction technique (PCR) was used to determine the resistance genes of the most five antimicrobial agents (sulfamethoxazol, tetracycline, erythromycin, ampicillin and nalidixic acid) showed resistance in the isolated Salmonella and E. coli from both migratory birds and broiler farms. DNA was extracted from the selected isolates and examined for the presence of (sulfamethoxazol; Sul1, tetracycline; TetA (A), erythromycin; ereA, ampicillin; blaTEM and nalidixic acid; qnrA). The oligonucleotide primers that used were supplied from Metabion (Germany) (Supplementary Table 1). Agarose gel (1.5%) with ethidium bromide (5 µL) (Invitrogen, UltraPure, Waltham, MA) was prepared using Tris − acetate − EDTA buffer (0.5 M). DNA ladder (100-bp; Promega) was pipetted into the first well of each gel, with the samples loaded in the other wells of the gel (8 µL), and gel electrophoresis was performed at 135 V for 20 min. Then, the bands were detected under UV light and were photographed using gel documentation system (Alpha Innotech, Biometra) [30].

Detection of genetic diversity by ERIC‑PCR

Four common Salmonella serotypes (S. Enteritidis, S. Kentucky, S. Typhimurium and S. Shangani) and 7 common E. coli serogroups (O26, O55, O91, O103, O128, O125 and O159) were selected and examined by ERICPCR technique to determine the similarity between the same serotypes. The extraction process was performed using QIAamp DNA mini kit (Qiagen- Germany- GmbH). The oligonucleotide primers that used were supplied from Metabion (Germany) (Supplementary Table 1). The gel was prepared and photographed as mentioned previously and the data was analyzed by the computer software.

ERIC fingerprinting data were transformed into a binary code depending on the presence or absence of each band. Dendrograms were generated by the unweighted pair group method with arithmetic average (UPGMA) and Ward’s hierarchical clustering routine. Cluster analysis and dendrogram construction were performed with SPSS, version 22 (IBM 2013) [32]. Similarity index (Jaccard / Tanimoto Coefficient and number of intersecting elements) was calculated using the online tool (https://planetcalc.com/1664/).

Statistical analysis

Microsoft Excel (Version 15.0) was used for the data recording and SPSS (Statistical Package for Social Science) (version 25) was used to perform data analysis.

Results

Overall, we isolated 298 (28.4%) of S. enterica strains belonging to 27 serovars from both migratory birds and poultry farms, including S. Typhimurium (n = 88), S. Enteritidis (n = 82), S. Kentucky (n = 45), S. Papuana (n = 3), S. Larochelle (n = 4), S. Alfort (n = 5), S. Shangani (n = 15), S. Tsevie (n = 7), S. Shubra (n = 4), S. Paratyphi A (n = 2), S. Heidelbergand (n = 8), S. Colindale (n = 1), S. Daula (n = 3), S. Bargny (n = 1), S. Infantis (n = 3), S, Inganda (n = 4), S. Angers (n = 1), S. Molade (n = 1), S. Newport (n = 3), S. Apeyeme (n = 2),S. Lexington (n = 1), S. Labadi (n = 1), S. Rechovot (n = 1), S. Tamale (n = 2), S. Virchow (n = 3), S. Wingrove (n = 2) and S. Montevideo (n = 6). There were 4 common serovars (S. Typhimurium, S. Enteritidis, S. Kentucky and S. Shangani) between migratory birds and poultry farms isolates. Regarding to the E. coli, we isolated 489 (46.6%) isolates of E. coli belonging to 24 serogroups, including O1 (n14), O2 (n = 8), O103 (n = 7), O124 (n = 7), O125 (n = 16), O128 (n = 39), O142 (n = 6), O144 (n = 13), O151 (n = 8), O158 (n = 2), O159 (n = 4), O166 (n = 14), O26 (n = 24), O55 (n = 11), O28 (n = 9), O6 (n = 6), O63 (n = 10), O86 (n = 16), O91 (n = 39), O146 (n = 7), O15 (n = 4), O163 (n = 5), O17 (n = 5) and O78 (n = 40).

The recorded results in Table 1 revealed that the prevalence of S. enterica was 28.4% (298 isolates out of 1050 sample) including 169 isolates from migratory birds and 129 isolates from the examined poultry farms. On the level of migratory birds, the prevalence of S. enterica was 21.3% in 2019, 24.7% in 2020, 28% in 2021, 16.7% in 2022 and 22% in 2023. On the farm level, Salmonella was isolated with 46.7% in 2019, 46.7% in 2020, 33.3% in 2021, 45% in 2022 and 43.3% in 2023. Regarding to the E. coli prevalence during the five years of the study was 46.6% (489 out of 1050) including 244 isolates from migratory birds and 255 isolates from poultry farms. On the level of migratory birds, it was 38.7% in 2019, 26.7% in 2020, 34% in 2021, 40.7% in 2022 and 22.7% in 2023. Meanwhile, the prevalence of E. coli in poultry farms was 78.3% in 2019, 83.3% in 2020, 76.7% in 2021, 86.7% in 2022 and 83.3% in 2023. The higher isolation was found in the year 2022 for both migratory bird with 40.7% and for farms with (86.7%). Meanwhile, the lower isolation was recorded from migratory birds in 2023 with 22.7%and from farms in 2021 with 76.7%. Another seven species were recorded; Citrobacter, Enterobacter, Klebsiella, Proteus, Providencia, Serratia, Hafnia and all of them were isolated from the collected samples in the years of study except for Hafnia that recorded only from migratory birds in 2019. The results in Table 1 revealed that Proteus and Klebsiella spp. were the most 2 species showed the highest isolation after Salmonella and E. coli was 26.7% in 2019 and 43.3% in 2020 from migratory birds and farms, respectively.

Table 1 Prevalence of Enterobacteriaceae isolates from migratory birds and broiler chicken farms
Full size table

Concerning to the results of the serotyping (Table, 2), the results revealed that S. Typhimurium, S. Kentucky and S. Enteritidis were the most predominant serotypes isolated from both migratory birds and farms. Four serotypes (S. Typhimurium, S. Kentucky, S. Enteritidis and S. Shangani) were recorded as common serotypes between migratory birds and farms. From (Table 3), 3 predominant E. coli serogroups (O91, O26, and O128) were reported from migratory birds and broiler chicken farms. E. coli O2 and O78 were isolated only from migratory birds. Seven serogroups (O91, O128, O26, O125, O55, O103 and O159) were common serogroups between migratory birds and farms samples.

Table 2 Prevalence of the isolated Salmonella serotypes in migratory birds and broiler chicken farms
Full size table
Table 3 Prevalence of the isolated E. Coli serogroup in migratory birds and broiler chicken farms
Full size table

The in vitro antimicrobial susceptibility of the recovered Salmonella and E. coli against 12 antimicrobial agents belonged to 8 different classes showed resistance in the majority of the examined isolates. From Table 4, the high level of resistances in Salmonella recovered from migratory birds during the study period were recorded in nalidixic acid (100%), tetracycline (31 to 100%), ampicillin (46.9 to 100%), sulfamethoxazol (68.8 to 100%), clindamycin (31.3 to 100%), and erythromycin (87.5 to 100%). Also, in the recovered Salmonella from farm, the highest resistance was found for nalidixic acid (100%), ampicillin (92.6 to 100%), sulfamethoxazol (85.7 to 100%), clindamycin (75 to 100%), and erythromycin (92.6–100%).

Table 4 Antimicrobial resistant pattern of the isolated salmonellae from migratory birds and broiler chicken farms
Full size table

From Table (5), the highest resistance in E. coli that isolated from migratory birds during the study period were recorded in nalidixic acid (70.6 to 100%) and clindamycin (70.7 to 91.8%), and erythromycin (98.3–100%). Meanwhile, high resistant E. coli that recovered from broiler farms was recorded for nalidixic acid (92 to 100%), clindamycin (78.7 to 100%), cefotaxime (85.1 to 100%) and erythromycin (94.2 to 100%).

Table 5 Prevalence of antimicrobial resistance of the isolated E. Coli from migratory birds and broiler chicken farms
Full size table

The majority of Salmonella (91.6%; 274 out of 298) and E. coli (92%; 450 out of 489) strains from both migratory birds and poultry farms showed MDR to the tested antimicrobial agents. The MDRI was reported in Salmonella isolated from migratory bird and farms which ranged from 0.17 to 1 and from 0.33 to 1, respectively. Regarding to E. coli, the MDRI in migratory birds ranged from 0.08 to 1 but in farms ranged from 0.33 to 1. On the level of farms, the different antimicrobial resistance patterns for Salmonella strains were 44 at which NA, E, AM, SXT, CL, T, CP was the most common one and for E. coli strains were 66 and E, CL, T, NA, CF, SXT, AM, CP, G, L, K, AK was the most common one (Supplementary Tables 3 &4). Meanwhile, on the level of migratory birds there were 50 different antimicrobial resistance patterns for Salmonella strains at which NA, E, AM, SXT was the most common pattern and 61 different patterns for E. coli strains at which E, CL, T, NA, CF, SXT was the most common one.

The recorded results in Table 6 demonstrated that the prevalence of Sul1, TetA (A), blaTEM, ere A and qnrA genes (Figs. 1, 2, 3, 4 and 5) in the resistant Salmonella strains from migratory birds were 96.7%, 82%, 41.3%, 94.7% and 96.3%, respectively, but its prevalence in broiler farms were 96.7%, 95.6%, 46.8%, 97.6% and 98.4%, respectively. Meanwhile, the prevalence of Sul1, TetA (A), blaTEM and qnrA genes in the resistant E. coli strains which isolated from migratory birds were 95.6%, 91.7%, 36.6%, 97.7% and 98.5%, respectively, and in broiler farms were 98.7%, 96.6%, 34.7%, 97.4% and 98.3%, respectively (Figs. 1, 2, 3, 4 and 5).

Fig. 1
figure 1

Representative full length of agarose gel electrophoresis of PCR products for E. coli (Lanes: 1–10) and Salmonella (Lanes: 11–20) isolates to detect Tet A gene in genomic DNA at 570 bp. Lane L: DNA ladder, P: Positive control, N: Negative control and Lanes: 2 to 5 were positive E. coli isolates from migratory birds in poultry farms. Lanes: 6 to 10 were positive E. coli isolates from migratory birds in live bird markets. Lanes: 11 to 15 were positive Salmonella isolates from migratory birds in poultry farms. Lanes: 16 to 20 were positive Salmonella isolates from migratory birds in live bird markets

Full size image
Fig. 2
figure 2

Representative full length of agarose gel electrophoresis of PCR products for E. coli (Lanes: 1–10) and Salmonella (Lanes: 11–20) isolates to detect Sul 1 gene in genomic DNA at 433 bp. Lane L: DNA ladder, P: Positive control, N: Negative control and Lanes: 1 to 5 except 4 were positive E. coli isolates from migratory birds in poultry farms. Lanes: 6 to 10 were positive E. coli isolates from migratory birds in live bird markets. Lanes: 12 to 15 were positive Salmonella isolates from migratory birds in poultry farms. Lanes: 16 to 20 were positive Salmonella isolates from migratory birds in live bird markets

Full size image
Fig. 3
figure 3

Representative full length of agarose gel electrophoresis of PCR products for E. coli (Lanes: 1–10) and Salmonella (Lanes: 11–20) isolates to detect qnrA gene in genomic DNA at 516 bp. Lane L: DNA ladder, P: Positive control, N: Negative control and Lanes: 1 to 5 were positive E. coli isolates from migratory birds in poultry farms. Lanes: 6 to 10 were positive E. coli isolates from migratory birds in live bird markets. Lanes: 11 to 15 were positive Salmonella isolates from migratory birds in poultry farms. Lanes: 16 to 20 were positive Salmonella isolates from migratory birds in live bird markets

Full size image
Fig. 4
figure 4

Representative full length of agarose gel electrophoresis of PCR products for E. coli (Lanes: 1–10) and Salmonella isolates (Lanes: 11–20) to detect ereA gene in genomic DNA at 420 bp. Lane L: DNA ladder, P: Positive control, N: Negative control and Lanes: 1 and 3 were positive E. coli isolates from migratory birds in poultry farms. Lanes: 7 and 10 were positive E. coli isolates from migratory birds in live bird markets. Lanes: 13 were positive Salmonella isolates from migratory birds in poultry farms. Lanes: 16 and 17 were positive Salmonella isolates from migratory birds in live bird markets

Full size image
Fig. 5
figure 5

Representative full length of agarose gel electrophoresis of PCR products for E. coli (Lanes: 1–10) and Salmonella isolates (Lanes: 11–20) to detect blaTEMgene in genomic DNA at 516 bp. Lane L: DNA ladder, P: Positive control, N: Negative control and Lanes: 1 to 5 were positive E. coli isolates from migratory birds in poultry farms. Lanes: 6 and 10 were positive E. coli isolates from migratory birds in live bird markets. Lanes: 11 to 15 were positive Salmonella isolates from migratory birds of poultry farms. Lanes: 16 and 20 were positive Salmonella isolates from migratory birds of live bird markets

Full size image
Table 6 Prevalence and distribution of resistance genes in the isolated Salmonella strains
Full size table

ERIC showed high genetic similarity between the bacterial strains. The electrophoretic profile of DNA fragments obtained from 28 E. coli strains (Figs. 6) and 16 Salmonella strains (Fig. 7) produced 1–4 bands for E. coli and 5–8 bands for Salmonella strains, whose size ranged from 2000 to 2950 bp for and from 100 to 2974, respectively. Salmonella strains were clustered into 2 groups including X1 (n = 12) and X2 (n = 4), also E. coli strains were clustered into 2 groups as X1 (n = 9) and X2 (n = 19). For E. coli, the genetic similarity was found between many isolates for example No. 22 & 19 with isolates No. 23&1 from migratory birds and broiler chicken farms, respectively (Fig. 8). There were 14 different ERIC types were found in the examined Salmonella isolates (Fig. 9). The similarity was found only between isolate No. 7 from migratory birds and isolates No. 4 & 6 from broiler chicken farms.

Fig. 6
figure 6

Gel electrophoresis of PCR products for 28 E. coli strains which produced 1–4 bands whose size ranged from 2000 to 2950 bp. Lanes; 1–10,14, 16, 23, 24, 26 &27 from farms and lanes; 11–13, 15, 17–22, 25 & 28 from migratory birds

Full size image
Fig. 7
figure 7

The electrophoretic profile of DNA fragments obtained from 16 Salmonella strains which produced 5–8 bands whose size ranged from 100 to 2974 bp. Lanes; 1,3, 4, 6, 8–12 & 16 from farms and lanes; 2, 5, 7& 13–15 from migratory birds

Full size image
Fig. 8
figure 8

Dendrogram representing genetic relationships between E. coli isolates from migratory birds (isolate ID 22, 19, 25, 28, 17, 15, 11, 12,13, 18, 20 & 21; which marked with red color) and broiler chicken farms (isolates ID 23,1, 24, 27,16, 26, 8, 4, 5, 6, 7, 14, 2, 3, 9 & 10; which marked with green color) based on ERIC-PCR fingerprints. Eleven ERIC profile represented by A–J and the isolates ID represented by 1–28

Full size image
Fig. 9
figure 9

Dendrogram representing genetic relationships between Salmonella isolates from migratory birds (isolate ID 7, 5, 13, 14, 15 & 2; which marked with red color) and broiler chicken farms (isolates ID 4, 6,3,1, 12, 11, 16, 9, 10 & 8; which marked with green color) based on ERIC-PCR fingerprints. Fourteen ERIC profile represented by A–N and the isolates ID represented by 1–16

Full size image

The heatmap (Fig. 10) showed that the most important pathogenic strains of Salmonella (Enteritidis, Typhimurium, Infantis and Kentucky) were isolated from migratory birds and nearly all the isolates were multidrug resistant with different MDR profiles. On the other hand, the heatmap (Fig. 11) showed that the most important pathogenic serotypes of E. coli including O1, O26 and O78 at which O78 were the predominant strain from migratory birds. All the isolates which were represented in the heatmap were multidrug resistant with different MDR profiles but, there were similarity between the strains of migratory birds and poultry chicken farms in these profiles for the same serotype.

Fig. 10
figure 10

Salmonella Enteritidis, S. Typhimurum, S. Kentaky and S. Infantis (the most important Pathogenic strains) which were isolated from migratory birds and broiler farms were selected and used to construct a heatmap. Clustering demonstrated predominant multidrug resistant strains were isolated from migratory birds and broiler farms

Full size image
Fig. 11
figure 11

E. coli O1, O26 and O78 (the most important Pathogenic serotypes) which were isolated from migratory birds and broiler farms were selected and used to construct a heatmap. Clustering demonstrated predominant multidrug resistant strains were isolated from migratory birds and broiler farms

Full size image

Discussion

Salmonella and E. coli are the most common pathogens affecting poultry, in the current study, the higher percentages of Salmonella isolation were reported from migratory birds in the year 2021 with 28% and from farms in 2019 and 2020 with 46.7% for each year. Some research studies that performed on migratory birds come in near similarity with the current study whereas Salmonella was isolated with 28.26% in Egypt [33] and 21.21% in Bangladesh [18].

The highest level of Salmonella from broiler farms was determined during 2019 and 2020 with 46.7% for each. Our findings were agreed with Al-baqir et al. [34] who isolated Salmonella from chickens in Egypt with 32.6%. Lower frequencies of Salmonella isolation were recorded by Leinyuy et al. [35] who isolated it with 7.14%, 13%, 11.50% and 18.78%, respectively. The serotyping of Salmonella in the current study revealed the detection of 3 predominant serotypes (S. Typhimurium, S. Kentucky and S. Enteritidis) from both migratory birds and farms which were common between migratory birds and farms in accordance with Al-baqir et al. [34]. On the other hand, we detected rare serovars such as.

E. coli was isolated from migratory birds with (38.7%, 2019), (26.7%, 2020), (34%, 2021), (40.7%, 2022) and (22.7%, 2023). Our findings concerned with E. coli detection in migratory birds agreed to some extent with Yuan et al. [36] who recovered E. coli with (34.7%) in China. In this study, E. coli was isolated from broiler farms with (78.3%, 2019), (83.3%, 2020), (76.7%, 2021), (86.7%, 2022) and (83.3%, 2023). A research study conducted by Tawakol and Younis [37] on migratory birds identified 4 serogroups (O125, O126, O158 and O86). From the migratory birds collected in Texas, Callaway et al. [38] isolated E. coli O157:H7 from 3.7% of samples. The observed results of E. coli recover from broiler farms in this study were in accordance with Ozaki et al. [39] who isolated O125, O1 and O6.

Another seven species of the Enterobacteriacae family were recorded in this study; Citrobacter, Enterobacter, Klebsiella, Proteus, Providencia, Serratia, Hafnia and all of them were isolated from migratory birds and farms except, Hafnia that was recorded only from migratory birds at 2019. The differentiation of these species showed the detection of Citrobacter freundii, Enterobacter aerogenes, Enterobacter agglomerans, Enterobacter cloacae, Klebsiella pneumoniae, Proteus mirabilis, Proteus vulgaris, Providencia rettgeri and Serratia mascerans (Supplementary Table 2). Our findings were nearly similar to Raza et al. [40] and Giacopello et al. [41]. The current results from broiler farms agreed with Leinyuy et al. [35] and Moawad et al. [17].

Regarding to the in vitro antimicrobial susceptibility testing, the majority of the recovered Salmonella and E. coli from both migratory birds and broiler farms exhibited higher resistance to nalidixic acid, ciprofloxacin, tetracycline, ampicillin, sulfamethoxazol, clindamycin and erythromycin and our results agreed Card et al. [7]. Misuse of antibiotics specialy quinolones as unspecific treatment, or in subtherapeutic doses for prophylaxis or as growth promoters in developing countries, enhance the generations of antibiotic resistance in bacteria [42].

Regarding to Salmonella from the farms, our findings agreed with Shalaby et al. [11] who recorded total resistance to ampicillin and erythromycin. A significant MDR of Salmonella was reported to ampicillin, gentamycin [34]. Majority of the isolated Salmonella from broiler farms and E. coli from both migratory birds and farms showed MDR to the tested antimicrobial agents. These findings agreed with Kamboh et al. [42] who distinguished MDR in Salmonella isolated from migratory birds and Tawakol and Younis [37] who reported MDR in E. coli isolates.

The detection of the resistance genes in this study (table, 6) showed that the highest prevalence of sul1, tetA (A), blaTEM and qnrA genes were recorded in the resistant Salmonella and E. coli isolates from migratory birds and broiler farms. Our findings were nearly similar to Sharif et al. [3] who reported TEM gene in 100% of S. enterica which isolated from wild migratory birds. Regarding to the broiler farms, our findings nearly agreed with Alam et al. [43] who reported tetA and blaTEM−1 in Salmonella isolates with 97.14% and 82.85%, respectively. blaTEM−1, tet (A) and sul1 genes were detected in E. coli isolates with prevalence of 36.4%, 80.5% and 6.8%, respectively from migratory wild birds in China [23]. Our results in harmony with that of Yapicier et al. [44] who reported tet (A) gene in 54.3% of E. coli isolated from wild birds in Turkey. Meanwhile, Islam et al. [6]. recorded tet (A), qnrA and blaTEM with 100%, 35.71% and 95.24%, respectively.

ERIC-PCR and the heatmap indicated to the genetic similarity between the MDR E. coli strains that isolated from both migratory birds and poultry farms also, there was genetic similarity between MDR Salmonella strains that isolated from both migratory birds and poultry farms. These findings highlighted the potential role of migratory birds as vectors to disseminate MDR Salmonella and E. coli to poultry farms. This is necessitating to keep migratory birds under continuous antimicrobial surveillance programs with the application of biosecurity measures for the prevention of migratory birds from the entrance to poultry farms [45].

Data availability

The datasets used and/or analyzed during the current study were provided within the manuscript and supplementary information files.

References

  1. Helmy YA, El-Adawy H, Abdelwhab EM. A Comprehensive Review of Common Bacterial, parasitic and viral zoonoses at the human-animal interface in Egypt. Pathogens. 2017;6:33. 10.3390.

    Article  PubMed  PubMed Central  Google Scholar 

  2. Hoath R. (2021): The birds of Egypt and the Middle East (1st edition). The American university in Cairo press, Egypt.

  3. Sharif M, Alam S, Fazal S, Kabir M, Shah A, Khan W, Khan MM, Khurshid A. Isolation and characterization of multidrug resistant beta-lactamase producing Salmonella enterica from wild migratory birds. Appl Ecol Environ Res. 2020;18(1):1407–18.

    Article  Google Scholar 

  4. Dhama K, Mahendran M, Tomar S. Pathogens transmitted by migratory birds: threat perceptions to Poultry Health and Production. Int J Poult Sci. 2008;7(6):516–25.

    Article  Google Scholar 

  5. Nabil NM, Yonis AE. (2019): Importance of migratory birds as a vector in spreading of Salmonella in Egypt in the period from November 2017 to March 2018. Assiut Vet. Med. J. Vol. 65 No. 161 April 2019, 104–115.

  6. Islam MS, Sobur MA, Rahman S, Ballah FM, Ievy S, Siddique MP, Rahman M, Kaf A, Rahman M, M. T. Detection of blaTEM, blaCTX M, blaCMY, and blaSHV genes among extended spectrum beta lactamase producing Escherichia coli isolated from migratory birds travelling to Bangladesh. Microb Ecol. 2022;83(4):942–50.

    Article  CAS  PubMed  Google Scholar 

  7. Card RM, Chisnall T, Begum R, Sarker MS, Hossain MS, Sagor MS, Mahmud MA, Uddin ASMA, Karim MR, Lindahl JF, Samad MA. Multidrug-resistant non-typhoidal Salmonella of public health significance recovered from migratory birds in Bangladesh. Front Microbiol. 2023;14:1162657.

    Article  PubMed  PubMed Central  Google Scholar 

  8. Hassan WH, Hassan HS, Hassan WMM, Shany SAS, Osman GSI. Identification and characterization of Salmonella species isolated from broiler chickens. J Vet Med Res. 2021;28(1):21–9.

    Article  CAS  Google Scholar 

  9. Zakaria S, Ahmed AI, Osman N, Nasef SA. Antimicrobial susceptibility to Salmonella and E. Coli isolates originated from broiler chickens in Luxor Governorate. Assiut Vet Med J. 2018;64(157):163–72.

    Article  Google Scholar 

  10. Ejeh FE, Lawan FA, Abdulsalam H, Mamman PH, Kwanashie CN. Multiple antimicrobial resistance of Escherichia coli and Salmonella species isolated from broilers and local chickens retailed along the roadside in Zaria, Nigeria. Sokoto J Veterinary Sci. 2017;15(3):45–53.

    Article  Google Scholar 

  11. Shalaby A, Ismail M, El-Sharkawy H. (2022): Isolation, identification, and genetic characterization of antibiotic resistance of Salmonella species isolated from chicken farms. J Trop Med, (1) 6065831.

  12. Dudhane RA, Bankar NJ, Shelke YP, Badge AK. (2023): The Rise of Non-typhoidal Salmonella Infections in India: Causes, Symptoms, and Prevention. Cureus. 2023;15(10):e46699. https://doiorg.publicaciones.saludcastillayleon.es/10.7759/cureus.46699. PMID: 38021876; PMCID: PMC10630329.

  13. Kiran S, Waheed A, Khan AA, Aziz M, Ayaz MM, Sheikh AS. Differentiation of Human and Migratory Water Fowl by Multiplex Escherichia coli Differential amplification technique (MECDAT) in South Punjab, Pakistan. J Trop Dis. 2018;6:264.

    Article  Google Scholar 

  14. Ibrahim RA, Cryer TL, Lafi Q, Abu Basha E, Good L, Tarazi YH. (2019): Identification of Escherichia coli from broiler chickens in Jordan, their antimicrobial resistance, gene characterization and the associated risk factors. BMC Veterinary Research (2019) 15:159.

  15. Ellakany HF, Abd-Elhamid HS, Ibrahim MS, Mostafa NS, Elbestawy AR, Ahmed R, Gado. Isolation, serotyping, pathogenicity and antibiotic sensitivity testing of Escherichia Coli from Broiler Chickens in Egypt. AJVS. 2019;61(2):45–51.

    Article  Google Scholar 

  16. Islam MS, Paul A, Talukder M, Roy K, Sobur MA, Ievy S, Nayeem MMH, Rahman S, Nazir KHMNH, Hossain MT, Rahman MT. Migratory birds travelling to Bangladesh are potential carriers of multi-drug resistant Enterococcus spp., Salmonella spp., and Vibrio Spp. Saudi J Biol Sci. 2021;28(2021):5963–70.

    Article  Google Scholar 

  17. Moawad AA, Hotzel H, Neubauer H, Ehricht R, Monecke S, Tomaso H, Hafez HM, Roesler U, ElAdawy H. (2018): Antimicrobial resistance in Enterobacteriaceae from healthy broilers in Egypt: emergence of colistin-resistant and extended-spectrum β-lactamase-producing Escherichia coli. Gut Pathog (2018) 10:39.

  18. ISO 6579-1. (2017): Microbiology of food and animal feeding stuff- horizontal method for detection of enumeration and serotyping Salmonella SPP. International standard. (First ed., 02-2017).

  19. American Public Health Association. Standard methods for examination of waters and wastewater. 14th ed. Washington: D. C; 1971.

    Google Scholar 

  20. Kauffman G. : Kauffmann white scheme. J Acta Path Microbiol Sci. 1974;61:385.

    Google Scholar 

  21. Lee MD, Nolan KL. (2008): A laboratory manual for the isolation and identification of avian pathogen. In: Zavala, L.D., Swayne, D.E., Glisson, J.R., Jackwood, M.W., Pearson, J.E. and Reed, W.M., editors. Editorial Board for the American Association of Avian Pathologists. 5th ed., Ch. 3. 953 College Station Road Athens, GA 30602 – 4875.

  22. Quinn PJ, Markey BK, Carter ME, Donnelly WJ, Leonard FC. Veterinary Microbiology and Microbial Disease. 1st ed. Cornwall, Great Britain: Blackwell Science Ltd.; 2002. pp. 43–122.

    Google Scholar 

  23. CLSI. (2020): Performance Standards for Antimicrobial Susceptibility Testing. 30th ed. CLSI supplement M100. Wayne, PA: Clinical and Laboratory Standards Institute.

  24. Magiorakos AP, Srinivasan A, Carey RB, Carmeli Y, Falagas ME, Giske CG, et al. Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance. Clin Microbiol Infect Official Publication Eur Soc Clin Microbiol Infect Dis. 2012;18:268–81. https://doiorg.publicaciones.saludcastillayleon.es/10.1111/j.1469-0691.2011.03570.x.

    Article  CAS  Google Scholar 

  25. Randall LP, Cooles SW, Osborn MK, Piddock LJV, Woodward MJ. Antibiotic resistance genes, integrons and multiple antibiotic resistance in thirty-five serotypes of Salmonella enterica isolated from humans and animals in the UK. J Antimicrob Chemother. 2004;53:208–16.

    Article  CAS  PubMed  Google Scholar 

  26. Robicsek A, Strahilevitz J, Jacoby GA, Macielag M, Abbanat D, Park CH, Bush K, Hooper DC. Fluoroquinolone modifying enzyme: a new adaptation of a common aminoglycoside acetyltransferase. Nat Med. 2006;12:83–8.

    Article  CAS  PubMed  Google Scholar 

  27. Nguyen MCP, Woerther P, Bouvet M, Andremont A, Leclercq R, Canu A. Escherichia coli as reservoir for macrolide resistance genes. Emerg Infect Dis. 2009;15:10.

    Article  Google Scholar 

  28. Colom K, PèrezJ, Alonso R, Fernández-Aranguiz A. LariňoE, Cisterna R. (2003):Simple and reliable multiplex PCR assay for detection of blaTEM,blaSHV and blaOXA–1 genes in Enterobacteriaceae. FEMS Microbiology Letters 223 (2003) 147–151.

  29. Ibekwe AM, Murinda SE, Graves AK. Genetic diversity and Antimicrobial Resistance of Escherichia coli from Human and Animal sources uncovers multiple resistances from human sources. Volume 6. PLoS ONE; 2011. 6e20819.

  30. Ha AJ, Siberio Perez LG, Kim TJ, Rahaman Mizan MF, Nahar S, Park SH, Chun HS, Ha SD. (2020): Identification and characterization of Salmonella spp. in mechanically deboned chickens using pulsed-field gel electrophoresis. Poult Sci. 2021;100(3):100961. doi: 10.1016/j.psj.2020.12.058. Epub 2020 Dec 24. PMID: 33518318; PMCID: PMC7936188.

  31. Versalovic J, Koeuth T, Lupski JR. Distribution of repetitive DNA sequences in eubacteria and application to fingerprinting of bacterial genomes. Nucleic Acids Res. 1991;19:24 6823–31.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Hunter PR. Reproducibility and indices of discriminatory power of microbial typing methods. J Clin Microbiol. 1990;28:1903–5.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Shalaby AG, M. EA, Nasef SA. Surveillance for Salmonella in migratory, feral and zoo birds. SCVMJ. 2014;XIX(1):2014–85.

    Google Scholar 

  34. Al-baqir AM, Hussein AH, Ghanem IA, Megahed MM. Characterization of Paratyphoid Salmonellae isolated from Broiler Chickens at Sharkia Governorate, Egypt. Zag Vet J. 2019;47(2):183–92.

    Article  Google Scholar 

  35. Leinyuy JF, Ali IM, Karimo O, Tume CB. (2022): Patterns of Antibiotic Resistance in Enterobacteriaceae Isolates from Broiler Chicken in the West Region of Cameroon: A Cross-Sectional Study. Canadian Journal of Infectious Diseases and Medical Microbiology, 2022, 4180336.

  36. Yuan Y, Liang B, Jiang BW, Zhu LW, Wang TC, Li YG, Liu J, Guo XJ, Ji X, Sun Y. Migratory wild birds carrying multidrug-resistant Escherichia coli as potential transmitters of antimicrobial resistance in China. PLoS ONE. 2021;16(12):e0261444. https://doiorg.publicaciones.saludcastillayleon.es/10.1371/journal.pone.0261444. PMID: 34910771; PMCID: PMC8673662.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Tawakol MM, Younis AE. Role of migratory birds in transmission of E. Coli infection to Commercial Poultry. AJVS. 2019;62(2):72–81. July 2019.

    Article  Google Scholar 

  38. Callaway TR, Edrington TS, Nisbet DJ. Isolation of Escherichia coli O157:H7 and Salmonella from Migratory Brown-Headed cowbirds (Molothrus ater), common grackles (Quiscalus quiscula), and cattle egrets (Bubulcus ibis). Foodborne Pathog Dis. 2014;11(10):791–4.

    Article  PubMed  Google Scholar 

  39. Ozaki H, Matsuoka Y, Nakagawa E, Murase T. Characteristics of Escherichia coli isolated from broiler chickens with colibacillosis in commercial farms from a common hatchery. Poult Sci. 2017;96:3717–24.

    Article  CAS  PubMed  Google Scholar 

  40. Raza S, Mohsin M, A. Madni W, Sarwar F, Saqib M, Aslam B. First Report of blaCTX-M-15-Type ESBL-Producing Klebsiella pneumoniae in wild migratory birds in Pakistan. EcoHealth. 2017;14:182–6.

    Article  PubMed  Google Scholar 

  41. Giacopello C, Foti M, Mascetti A, Grosso F, Ricciardi D, Fisichella V, Piccolo FL. (2016): Antimicrobial resistance patterns of Enterobacteriaceae in European wild bird species admitted in a wildlife rescue centre. Veterinaria Italiana 2016, 52 (2), 139–144.

  42. Kamboh AA, Shoaib M, Abro SH, Khan MA, Malhi KK, Yu S. Antimicrobial Resistance in Enterobacteriaceae isolated from liver of commercial broilers and backyard chickens. J Appl Poult Res. 2018;27:627–34.

    Article  CAS  Google Scholar 

  43. Alam SB, Mahmud M, Akter R, Hasan M, Sobur A, Nazir KNH, Noreddin A, Rahman T, ElZowalaty ME, Rahman M. (2020): Molecular Detection of Multidrug Resistant Salmonella Species Isolated from Broiler Farm in Bangladesh. Pathogens 2020, 9, 201; https://doiorg.publicaciones.saludcastillayleon.es/10.3390/pathogens9030201

  44. Yapicier SO, Hesna Kandir E, Öztürk D. Antimicrobial Resistance of E. Coli and Salmonella isolated from Wild Birds in a Rehabilitation Center in Turkey. Arch Razi Inst. 2022;77(1):257–67.

    Google Scholar 

  45. Tawakol MM, Nabil NM, Samir A, Yonis AE, Shahein MA, Elsayed MM. The potential role of migratory birds in the transmission of pathogenic Campylobacter species to broiler chickens in broiler poultry farms and live bird markets. BMC Microbiol. 2023;23(1):1–11.

    Article  Google Scholar 

Download references

Funding

Open access funding provided by The Science, Technology & Innovation Funding Authority (STDF) in cooperation with The Egyptian Knowledge Bank (EKB). The authors declare that no funds, grants, or other support were received during the preparation of this manuscript.

Author information

Authors and Affiliations

  1. Reference Laboratory for Veterinary Quality Control on Poultry Production, Animal Health Research Institute (AHRI), Agricultural Research Center (ARC), Nadi El-Seid Street, Dokki, Giza, 12618, Egypt

    Maram M. Tawakol, Nehal M. Nabil, Abdelhafez Samir, Heba M. Hassan, Reem M. Reda & Ola Abdelaziz

  2. Animal Health Research Institute (AHRI), Agricultural Research Center (ARC), Hurghada, Egypt

    Sahar Hagag

  3. Department of Hygiene and Zoonoses, Faculty of Veterinary Medicine, Mansoura University, Mansoura, 35516, Egypt

    Mona M. Elsayed

Authors
  1. Maram M. Tawakol
    View author publications

    You can also search for this author inPubMed Google Scholar

  2. Nehal M. Nabil
    View author publications

    You can also search for this author inPubMed Google Scholar

  3. Abdelhafez Samir
    View author publications

    You can also search for this author inPubMed Google Scholar

  4. Heba M. Hassan
    View author publications

    You can also search for this author inPubMed Google Scholar

  5. Reem M. Reda
    View author publications

    You can also search for this author inPubMed Google Scholar

  6. Ola Abdelaziz
    View author publications

    You can also search for this author inPubMed Google Scholar

  7. Sahar Hagag
    View author publications

    You can also search for this author inPubMed Google Scholar

  8. Mona M. Elsayed
    View author publications

    You can also search for this author inPubMed Google Scholar

Contributions

M. M. T., N. M. N. and M. M. E. conceived the study and was involved in the design and coordination of the study. M. M. T., N. M. N., A. S., H. M. H., R. M. R., O. A., S. H., and M. M. E. were involved in practical part. M. M. T., N. M. N. and M. M. E. were involved in data analysis, manuscript drafting, and editing. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Mona M. Elsayed.

Ethics declarations

Ethical approval and consent to participate

This protocol was performed by following the animal ethics guidelines and approved by the Medical Research Ethics Committee of Mansoura University. All methods were carried out in accordance with relevant guidelines and regulations. Informed consent was obtained from the poultry farms owners for the use of their animals in the study prior to study commencement. All methods for samples collection were carried out according to the standard guidelines.

Competing interests

The authors declare no competing interests.

Consent for publication

Not applicable.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary Material 1

Supplementary Material 2

Rights and permissions

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Tawakol, M.M., Nabil, N.M., Samir, A. et al. Role of migratory birds as a risk factor for the transmission of multidrug resistant Salmonella enterica and Escherichia coli to broiler poultry farms and its surrounding environment. BMC Res Notes 17, 314 (2024). https://doiorg.publicaciones.saludcastillayleon.es/10.1186/s13104-024-06958-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doiorg.publicaciones.saludcastillayleon.es/10.1186/s13104-024-06958-7

Keywords

BMC Research Notes

ISSN: 1756-0500

Contact us