scholarly journals Bacteriophage activity against and characterisation of avian pathogenic Escherichia coli isolated from colibacillosis cases in Uganda

PLoS ONE ◽  
2020 ◽  
Vol 15 (12) ◽  
pp. e0239107
Author(s):  
George Kazibwe ◽  
Phionah Katami ◽  
Ruth Alinaitwe ◽  
Stephen Alafi ◽  
Ann Nanteza ◽  
...  

Avian Pathogenic Escherichia coli (APEC) cause colibacillosis leading to significant economic losses in the poultry industry. This laboratory-based study aimed at establishing stocks of avian pathogenic Escherichia coli lytic bacteriophages, for future development of cocktail products for colibacillosis management. The study determined the antibiotic susceptibility; phylogenetic categories, occurrence of selected serotypes and virulence genes among Escherichia coli stock isolates from chicken colibacillosis cases; and evaluated bacteriophage activity against the bacteria. Escherichia coli characterization was done through phenotypic and multiplex PCR methods. Bacteriophage isolation and preliminary characterization was achieved using the spot assay and overlay plating techniques. Fifty-six (56) isolates were phenotypically confirmed as E. coli and all exhibited resistance to at least one antimicrobial agent; while multi-drug resistance (at least three drugs) was encountered in 50 (89.3%) isolates. The APEC isolates mainly belonged to phylogroups A and D, representing 44.6% and 39.3%, respectively; whereas serotypes O1, O2 and O78 were not detected. Of the 56 isolates, 69.6% harbored at least one virulence gene, while 50% had at least four virulence genes; hence confirmed as APEC. Virulence genes, ompT and iutA were the most frequent in 33 (58.9%) and 32 (57.1%) isolates respectively; while iroN least occurred in 23 (41.1%) isolates. Seven lytic bacteriophages were isolated and their host range, at 1×108 PFU/ml, varied from 1.8% to 17.9% of the 56 APEC isolates, while the combined lytic spectrum was 25%. Phage stability was negatively affected by increasing temperatures with both UPEC04 and UPEC10 phages being undetectable at 70°C; whereas activity was detected between pH 2 and 12. The high occurrence of APEC isolates resistant against the commonly used antibiotics supports the need for alternative strategies of bacterial infections control in poultry. The low host range exhibited by the phages necessitates search for more candidates before in-depth phage characterization and application.

Author(s):  
Joshua Mbanga ◽  
Yvonne O. Nyararai

Colibacillosis, a disease caused by avian pathogenic Escherichia coli (APEC), is one of the main causes of economic losses in the poultry industry worldwide. This study was carried out in order to determine the APEC-associated virulence genes contained by E. coli isolates causing colibacillosis in chickens. A total of 45 E. coli isolates were obtained from the diagnostics and research branch of the Central Veterinary Laboratories, Bulawayo, Zimbabwe. These isolates were obtained from chickens with confirmed cases of colibacillosis after postmortem examination. The presence of the iutA, hlyF, ompT, frz, sitD, fimH, kpsM, sitA, sopB, uvrY, pstB and vat genes were investigated by multiplex polymerase chain reaction (PCR) assay. Of the 45 isolates, 93% were positive for the presence of at least one virulence gene. The three most prevalent virulence genes were iutA (80%), fimH (33.3%) and hlyF (24.4%). The kpsM, pstB and ompT genes had the lowest prevalence, having been detected in only 2.2% of the isolates. All 12 virulence genes studied were detected in the 45 APEC isolates. Virulence gene profiles were constructed for each APEC isolate from the multiplex data. The APEC isolates were profiled as 62.2% fitting profile A, 31.1% profile B and 6.7% profile C. None of the isolates had more than seven virulence genes. Virulence profiles of Zimbabwean APEC isolates are different from those previously reported. Zimbabwean APEC isolates appear to be less pathogenic and may rely on environmental factors and stress in hosts to establish infection.


2019 ◽  
Vol 7 ◽  
pp. 905-911
Author(s):  
Marilena Burtan ◽  
Virgilia Popa ◽  
Maria Rodica Gurau ◽  
Doina Danes

Introduction: Colibacillosis in poultry is determined by avian pathogenic Escherichia coli (APEC) and represents an important source of economic losses  in the poultry industry. APEC’s pathogenicity relies on the presence and expression of different virulence factors. The genes ompA , iss  and  fimH, encoding the outer membrane protein, the protein inducing resistance to complement and the synthesis of type 1 fimbria are present in APEC strains. Objective: Escherichia coli strains isolated from layers were analysed to assess the pathotype they belong to. Methods: In order to detect the three genes associated with APEC strains, 16 E. coli isolates were investigated for virulence associated genes ompA, iss and fimH, using multiplex PCR. Results: From the 16 E.coli strains submitted, multiplex PCR assessment revealed that 14 (87.5%) of the E. coli strains isolated contained at least one virulence gene, while 2 (12.5%) strains did not harbour any of the virulence genes tested. The fimH gene was noted in 13 (81.25%) of the strains tested, the ompA gene has been present in 12 (75%) strains and the iss gene was present in 9 (56.25%) strains. Eight (50%) strains were found to present all three investigated genes. Conclusion: Presence of these genes is a strong indicatory to consider those strains as belonging to the APEC pathotype.


2020 ◽  
Author(s):  
George Kazibwe ◽  
Phionah Katami ◽  
Ruth Alinaitwe ◽  
Stephen Alafi ◽  
Ann Nanteza ◽  
...  

AbstractA laboratory-based study aimed at establishing a stock of avian pathogenic Escherichia coli (APEC) lytic bacteriophages, for future development of cocktail products for controlling colibacillosis as well as minimizing use of antimicrobial drugs in the poultry production systems in Uganda. Specifically, the study determined the antibiotic susceptibility; phylogenetic categories, occurrence of selected virulence genes among Escherichia coli stock isolates from cases of chicken colibacillosis; and isolation of specific bacteriophages. Fifty six isolates were confirmed as E. coli by standard phenotypic tests. All the 56 (100%) isolates were resistant to at least one antibiotic while 50 (89.3%) isolates were resistant to at least three classes of antimicrobial drugs and were therefore designated as multi-drug resistant. Phylogenetically, APEC isolates mainly belonged to phylogroups A and D which represented 44.6% and 39.3%, respectively. Virulence genes, ompT and iutA were the most frequent with 33 (58.9%) and 32 (57.1%) isolates respectively; while iroN least occurred in 23 (41.1%) isolates. Of the 56 isolates, 69.6% harbored at least one virulence gene, while 50% had at least four virulence genes; hence confirmed as APEC. None of the isolates belonged to the selected serotypes O1, O2 and O78. Seven specific bacteriophages were isolated and their host range, varied from 1.8% to 17.9% (n=56 APEC isolates), while the combined lytic spectrum of all the phages was 25%. Phage stability was negatively affected by increasing temperatures with both UPEC04 and UPEC10 phages becoming undetectable at 70°C; however activity was detected between pH 2 and 12. The high occurrence of APEC isolates with resistance against the commonly used antibiotics supports the need for alternative strategies of bacterial infections control in poultry. The low host range exhibited by the phages calls for search for more candidates before more in-depth studies are done for phage characterization and application.


2019 ◽  
Vol 59 (2) ◽  
pp. 338
Author(s):  
S. N. Magray ◽  
S. A. Wani ◽  
Z. A. Kashoo ◽  
M. A. Bhat ◽  
S. Adil ◽  
...  

The present study has determined the serological diversity, virulence-gene profile and in vitro antibiogram of avian pathogenic Escherichia coli (APEC) isolates from broiler chickens in India suspected to have died of colibacillosis. The virulence-gene profile of APEC was compared with that of the Escherichia coli isolates from faeces of apparently healthy chickens, called avian faecal E. coli (AFEC). In total, 90 representative isolates of APEC and 63 isolates of AFEC were investigated in the present study. The APEC were typed into 19 serogroups, while some isolates were rough and could not be typed. Most prevalent serogroup was O2 (24.44%). Among the eight virulence genes studied, the prevalence of seven genes (iss, iucD, tsh, cva/cvi, irp2, papC and vat) was significantly higher in APEC than in AFEC isolates. However, there was no significant difference between APEC and AFEC isolates for possession of astA gene. The most frequent gene detected among the two groups of organisms was iss, which was present in 98.88% and 44.44% of APEC and AFEC isolates respectively. The in vitro antibiogram showed that the majority (96.6%) of APEC isolates were resistant to tetracycline, while 82.2% were resistant to cephalexin, 78.8% to cotrimoxazole, 68.8% to streptomycin and 63.3% to ampicillin. However, most of them (84.45%) were sensitive to gentamicin. Thus, it is concluded that APEC from the broiler chickens carried putative virulence genes that attributed to their pathogenicity. Furthermore, the majority of APEC isolates were found to be multi-drug resistant, which, in addition to leading treatment failures in poultry, poses a public health threat.


2021 ◽  
Vol 7 (1) ◽  
pp. 967-972
Author(s):  
Farzana Ehetasum Hossain ◽  
Saiful Islam ◽  
Md Aminul Islam ◽  
Shariful Islam ◽  
Firoz Ahmed

Avian colibacillosis, caused by avian pathogenic Escherichia coli (APEC), is one of the major infectious diseases of poultry that bring about great economic loss for the Bangladesh poultry industry. The present study aimed to determine the virulence genes of avian pathogenic Escherichia coli (APEC) from cases of colibacillosis in poultry at the Noakhali district of Bangladesh. Currently, virulence-associated gene profiles of APEC isolates were investigated by polymerase chain reaction (PCR). A total of 24 (twenty-four) Escherichia coli isolates were collected and presumptively identified from 8 (eight) colibacillosis cases from 4 commercial broiler poultry farms (2 broilers per farm) in Noakhali, Bangladesh. The pathogenesis of Escherichia coli involves a wide range of different virulence genes. At this point, four virulence genes, iutA, hlyF, iroN, and iss were detected by PCR analysis. It has been observed that iutA, iss, hlyF, and iroN genes were found in 7(29.16%), 20(83.33%), 22(91.66%), and 24(100%) APEC isolates respectively. Furthermore, out of the twenty-four APEC isolates, six (25%) isolates had four virulence genes, fourteen (58.33%) isolates carried at least three virulence genes, three (12.5%) isolates carried two genes and one (4.16%) isolates had one virulence gene. Most importantly. six types of virulence gene profiles existed within the APEC isolates from which profile number 3 (hlyF, iroN, iss) having 13 (54.16%) isolates were predominant. The occurrence of APEC isolates of this region which is responsible for avian colibacillosis cases can be a matter of concern from the public health point of view. Future investigations will be able to utilize these virulence genes to identify APEC in Bangladesh helping in the diagnosis and prevention of colibacillosis in poultry. Bioresearch Commu. 7(1): 967-972, 2021 (January)


2006 ◽  
Vol 188 (16) ◽  
pp. 5975-5983 ◽  
Author(s):  
Timothy J. Johnson ◽  
Sara J. Johnson ◽  
Lisa K. Nolan

ABSTRACT Avian pathogenic Escherichia coli (APEC), an extraintestinal pathogenic E. coli causing colibacillosis in birds, is responsible for significant economic losses for the poultry industry. Recently, we reported that the APEC pathotype was characterized by possession of a set of genes contained within a 94-kb cluster linked to a ColV plasmid, pAPEC-O2-ColV. These included sitABCD, genes of the aerobactin operon, hlyF, iss, genes of the salmochelin operon, and the 5′ end of cvaB of the ColV operon. However, the results of gene prevalence studies performed among APEC isolates revealed that these traits were not always linked to ColV plasmids. Here, we present the complete sequence of a 174-kb plasmid, pAPEC-O1-ColBM, which contains a putative virulence cluster similar to that of pAPEC-O2-ColV. These two F-type plasmids share remarkable similarity, except that they encode the production of different colicins; pAPEC-O2-ColV contains an intact ColV operon, and pAPEC-O1-ColBM encodes the colicins B and M. Interestingly, remnants of the ColV operon exist in pAPEC-O1-ColBM, hinting that ColBM-type plasmids may have evolved from ColV plasmids. Among APEC isolates, the prevalence of ColBM sequences helps account for the previously observed differences in prevalence between genes of the “conserved” portion of the putative virulence cluster of pAPEC-O2-ColV and those genes within its “variable” portion. These results, in conjunction with Southern blotting and probing of representative ColBM-positive strains, indicate that this “conserved” cluster of putative virulence genes is primarily linked to F-type virulence plasmids among the APEC isolates studied.


2019 ◽  
Vol 7 (1) ◽  
pp. 4-8
Author(s):  
Payam Haghighi Khoshkhoo ◽  
Hadi Pourtaghi ◽  
Gita Akbariazad ◽  
Saeed Mokhayeri

Background: Avian pathogenic Escherichia coli (APEC) causes economic losses in the chicken industry worldwide. Objective: In this study, virulence-associated gene profiles of APEC isolates were investigated by polymerase chain reaction (PCR). Materials and Methods: A total of 60 Escherichia coli isolates were collected from 60 colibacillosis cases from 30 broiler poultry farms in Alborz, Tehran, and Golestan provinces, Iran. After identification by biochemical tests, DNA was extracted by boiling method and 5 virulence-associated genes including: iutA, hlyF, iroN, ompT, and iss were detected by 2 multiplex PCR protocols. Results: Of the 60 APEC isolates, 26 (43.3%) isolates had at least three virulence genes from which 12 (20%) isolates were positive for all 5 virulence genes, whereas 34 (56.6%) carried no investigated virulence genes. Presence of iutA, hlyF, iroN, ompT, and iss genes in the APEC isolates were 17 (28.3%), 17 (28.3%), 24 (40%), 26 (43.3%), and 23 (38.3%), respectively. Conclusion: According to the results, four different virulence-associated gene profiles were seen in isolates, from which profile 1 with 12 (20%) isolates was predominant. These findings were in agreement with the previous reports.


2021 ◽  
Author(s):  
Alreem Johar ◽  
Najlaa Al-Thani ◽  
Sara Al-Hadidi ◽  
Elyes Dlissi ◽  
Mahmoud Mahoud ◽  
...  

Introduction: Avian Pathogenic Escherichia coli (APEC) is the contributing agent behind the avian infectious disease colibacillosis, which causes substantial fatalities in poultry industries that significantly impact the economy and food safety. Several virulence genes have been shown to be concomitant with the extra-intestinal survival of APEC. This study investigates the antibiotic resistance patterns and APEC‐associated virulence genes content in Escherichia coli (E. coli) isolated from non‐healthy and healthy broiler chickens from a commercial poultry farm in Qatar. Material and Methods: 158 E. coli strains were isolated from 47 chickens from five different organs (air sac, cloacal, kidney, liver, and trachea). Genomic DNA was extracted from E. coli using the QIAamp Pathogen Mini Kit. Multiplex PCR was executed to detect tsh, iucD, ompT, hlyF, iroN, iss, vat, cvi/cva genes associated with PPEC. Antibiotic susceptibility testing was performed using the standard Kirby-Bauer disk and E-test. Amplified virulence genes detected were sequenced and analyzed. Graph Pad version 8 and PAST software version 4.03 were used for statistical and clustering analysis. The chi-square test was performed on all data to compare the antibiotic resistance and virulence gene patterns between non-healthy and healthy chicken samples Results: 65% of the isolated bacteria were APEC strains containing five or more virulence genes, and 34% were non‐pathogenic E. coli (NPEC) strains. The genes ompT, hlyF, iroN, tsh, vat, iss, cvi/cva, and iucD were significantly prevalent in all APEC strains. E. coli isolates showed 96% resistance to at least one of the 18 antibiotics, with high resistance to ampicillin, cephalothin, ciprofloxacin, tetracycline, and fosfomycin. Conclusions: Our findings indicate high antibiotic resistance prevalence in non-healthy and healthy chicken carcasses. Such resistant E. coli can spread to humans. Hence, special programs are required to monitor the use of antibiotics in chicken production in Qatar.


2019 ◽  
Vol 63 (3) ◽  
pp. 317-320 ◽  
Author(s):  
Xiaojing Xu ◽  
Qing Sun ◽  
Lixiang Zhao

Abstract Introduction Avian pathogenicEscherichia coli (APEC) causes serious colibacillosis and significant economic losses. Data on profiles of virulence factors and antibiotic resistances among APEC strains are crucial to the control of infection. In this study, strains were isolated from eastern China, and the prevalence of virulence factors and distribution of antibiotic resistance were determined. Material and Methods APEC strains were isolated and characterised by PCR for O serogroups, virulence factor genes, antibiotic resistance, and phylogenetic groups. Results O78 was the most prevalent serogroup and type A was the most frequent phylogenetic group. ThefimH,feoB, andiron genes were the most prevalent among the isolates. All isolates were multiresistant, and all strains were resistant to ampicillin and tetracycline, which are widely used in the poultry industry in China. Conclusion This study provided important data on the presence of virulence genes and antibiotic resistance profiles of APEC from poultry farms in eastern China.


Antibiotics ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 564
Author(s):  
Alreem Johar ◽  
Najlaa Al-Thani ◽  
Sara H. Al-Hadidi ◽  
Elyes Dlissi ◽  
Mahmoud H. Mahmoud ◽  
...  

Avian Pathogenic Escherichia coli (APEC) is the contributing agent behind the avian infectious disease colibacillosis, which causes substantial fatalities in poultry industries that has a significant impact on the economy and food safety. Several virulence genes have been shown to be concomitant with the extraintestinal survival of APEC. This study investigates the antibiotic resistance patterns and APEC-associated virulence genes content in Escherichia coli isolated from non-healthy and healthy broiler chickens from a commercial poultry farm in Qatar. A total of 158 E. coli strains were isolated from 47 chickens from five different organs (air sac, cloacal, kidney, liver, and trachea). Based on genetic criteria, 65% were APEC strains containing five or more virulence genes, and 34% were non-pathogenic E. coli (NPEC) strains. The genes ompT, hlyF, iroN, tsh, vat, iss, cvi/cva, and iucD were significantly prevalent in all APEC strains. E. coli isolates showed 96% resistance to at least one of the 18 antibiotics, with high resistance to ampicillin, cephalothin, ciprofloxacin, tetracycline, and fosfomycin. Our findings indicate high antibiotic resistance prevalence in non-healthy and healthy chicken carcasses. Such resistant E. coli can spread to humans. Hence, special programs are required to monitor the use of antibiotics in chicken production in Qatar.


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