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Immuno-Bioinformatics Study of Autotransporter Protein, Antigen 43, in Enterotoxigenic Escherichia coli Isolated From Calves
2019
Ghorbanpour, Reyhaneh | Nikbakhat Brujeni, Gholamraza | Jalali, Seyed Amir Hossein
BACKGROUND: Extensive effort is focused on identifying genomic conserved antigens in development of effective vaccine against Enterotoxigenic Escherichia coli. Antigen 43 is one of the members of a large secreted protein family named autotransporters in the E.coli and other gram negative bacteria. Autotransporter proteins have a similar conserved structure. Some of them are recognized during both experimental and naturally occurring ETEC infections. Antigen 43 is represented as a potential target in vaccine development because of its virulence functions such as cell aggregation, biofilm formation and its presence in convalescent sera from human patients with ETEC diarrhea. OBJECTIVES: In this study, we carefully investigate antigenic structure and immunogenicity of the Antigen 43 protein of strain 510 E.coli isolated from calves by experimental methods and immunoinformatics tools. METHODS: Amino acid sequence, physico-chemical parameters, stability, secondary and tertiary protein structure, the ability of induction the B and T cell immune responses by having the effective epitopes and also the allergenicity assessment were analyzed. RESULTS: In this study, we identified 15 peptide sequences that can potentially induce B and T cell immune responses and finally, 9 of them were introduced as antigens. CONCLUSIONS: The results of in-silico analysis on this protein suggested that it can be used in bovine colibacillosis vaccine development.
Show more [+] Less [-]Presence of Two Genes Involved in Serum Resistance of Escherichia coli Isolated From Healthy Ostriches in Comparison With Infected Poultry by Colibacillosis
2019
Hosseini, Afsaneh | Salari, Saeed | Rashki, Ahmad | Jahantigh, Mohammad
BACKGROUND: The mechanism of pathogenesis and the role of virulence factors of avian pathogenic E. coli is still ill-defined. The ostrich industry is expanding, resulting in the interaction between poultry and ostrich. It is reported that the investigation of iss and bor virulence genes together, due to their structural and functional similarities, is valuable. Objectives: The investigation and comparison of presence of two genes involved in serum resistance, iss and bor, in E. coli isolated from apparently healthy ostriches and poultry with colibacillosis. Methods: As a cross-sectional study, E. coli was recovered from fecal samples of apparently healthy ostriches and poultry with colibacillosis, and iss and bor genes were screened and compared via PCR in E. coli isolates. Results: iss frequencies, with no statistical difference, were 50% and 64.4% in E. coli isolated from apparently healthy ostriches and poultry with colibacillosis, respectively (P>0.05). 31.8% and 15.6% of E. coli isolated from apparently healthy ostriches and poultry with colibacillosis were positive for bor, respectively, with no statistical difference (P>0.05). 11.1% of isolates from colibacillosis and 18.2% of isolates from apparently healthy ostriches feces, with no statistical difference, were positive for both genes (P>0.05). Conclusions: Equal statistical distribution of both genes, bor and iss, between apparently healthy ostriches and poultry with colibacillosis and the health level of studied ostriches indicated that E. coli isolated from ostrich, probably employs other virulence factors instead of bor and iss to establish a disease. This hypothesis needs to examine more virulence genes in ostrich-origin E. coli. In addition, the ostrich feces could be introduced as a source of iss and bor genes.
Show more [+] Less [-]In silico analysis of virulence associated genes in genomes of Escherichia coli strains causing colibacillosis in poultry
2017
Kołsut, Joanna | Borówka, Paulina | Marciniak, Błażej | Wójcik, Ewelina | Wojtasik, Arkadiusz | Strapagiel, Dominik | Dastych, Jarosław
Introduction: Colibacillosis – the most common disease of poultry, is caused mainly by avian pathogenic Escherichia coli (APEC). However, thus far, no pattern to the molecular basis of the pathogenicity of these bacteria has been established beyond dispute. In this study, genomes of APEC were investigated to ascribe importance and explore the distribution of 16 genes recognised as their virulence factors. Material and Methods: A total of 14 pathogenic for poultry E. coli strains were isolated, and their DNA was sequenced, assembled de novo, and annotated. Amino acid sequences from these bacteria and an additional 16 freely available APEC amino acid sequences were analysed with the DIFFIND tool to define their virulence factors. Results: The DIFFIND tool enabled quick, reliable, and convenient assessment of the differences between compared amino acid sequences from bacterial genomes. The presence of 16 protein sequences indicated as pathogenicity factors in poultry resulted in the generation of a heatmap which categorises genomes in terms of the existence and similarity of the analysed protein sequences. Conclusion: The proposed method of detection of virulence factors using the capabilities of the DIFFIND tool may be useful in the analysis of similarities of E. coli and other sequences deriving from bacteria. Phylogenetic analysis resulted in reliable segregation of 30 APEC strains into five main clusters containing various virulence associated genes (VAGs).
Show more [+] Less [-]In silico analysis of virulence associated genes in genomes of Escherichia coli strains causing colibacillosis in poultry
2017
Kołsut Joanna | Borówka Paulina | Marciniak Błażej | Wójcik Ewelina | Wojtasik Arkadiusz | Strapagiel Dominik | Dastych Jarosław
Introduction: Colibacillosis – the most common disease of poultry, is caused mainly by avian pathogenic Escherichia coli (APEC). However, thus far, no pattern to the molecular basis of the pathogenicity of these bacteria has been established beyond dispute. In this study, genomes of APEC were investigated to ascribe importance and explore the distribution of 16 genes recognised as their virulence factors.
Show more [+] Less [-]Common pathogens diagnosed in pig samples from year 2014 to 2017 by Veterinary Research Institute
2018
Aisya Naama T. | Azizah D. | Masrin A. | Nurulaini R. | Roslina H. | Zurin Azlin M. J. | Chandrawathani, P. | Sohayati A. R. | Nurul Fatiha A. S. | Roseliza R.
A total of 23,322 specimens collected between 2014 and 2017, froma total of 2,592 cases, were received in Veterinary Research Institute, Ipoh (VRI) from various states in Malaysia and testedfor common bacterial, viral, and parasitic diseases in pigs. The highest occurrence of isolated bacteria from 771 samples whichtested positive were Salmonella (47.38%) and Escherichia coli (15.68%), followed by Staphylococcus (6.62%), Streptococcus (5.57%), Klebsiella pneumonia (4.88%), Pseudomona (3.38%), Acinetobacter (3.14%), Aeromonas (2.79%), Enterobacter (2.44%), one each of Bacillus and Pasteurella multocida (1.74%), Enterococcus (1.39%) and Corynebacterium (1.05%). 1.74% of each bacteria detected were Moxarella, Aspergillus, Burkholderia andChromobacterium. Positive samples tested by ELISA was Japanese encephalitis virus (JEV) (9.15%), Aujezsky disease virus (ADV)(5.37%), porcine cirvo-virus-2 (PCV2) (5.09%) and porcine reproductive and respiratory syndrome virus (PRRSV) (4.52%). Positive amples tested by the molecular test wasPCV2 (1.62%), PRRSV (1.32%) and classical swine fever virus (CSFV) (0.4%). Serology tests were conducted on 11,305 samplesand reported positive for Brucella suis (15.32%), Brucella abortus (0.62%), Brucella melitensis (0.85%), and melioidosis (0.05%). Parasitology analyses on 99 samples. revealed presence of 10.1% coccidia and 1% each of helminths and Sarcocystis. Within the 4-year period, there were no positive samples for porcine parvovirus (PPV), Nipah virus, swine influenza virus (SIV), and bacteria of Johne’s disease and leptospirosis. Continuous assessment is required to establish a comprehensive baseline data of swine diseases in Malaysia.
Show more [+] Less [-]Phylogenetic grouping and virulence gene profiles of Escherichia coli isolated from chicken
2017
Ramlan M. | S. Khairani Bejo | Khoo, E. | Roseliza R. | Zunita Z.
Colibacillosis is a disease caused by avian pathogenic E. coli (APEC) and is one of the principle cause of morbidity and mortality in poultry worldwide which is represented by a complex syndrome characterized by multiple organ lesions. This study was carried out to determine phylogenetic grouping and virulenceassociated genes contained by E. coli isolates which is related in causing disease in chicken. E. coli isolates obtained from clinical cases of Veterinary ResearchInstitute were re-identified by conventional methods. Phylogenetic grouping of the isolates was determined by triplex polymerase chain reaction (PCR), and the presence of eight virulence genes were identified by multiplex PCR. A total of 125 E. coli isolates were subjected toanalysis of phylogenetic background and virulence associated genes profiling. Phylogenetic analysis demonstrated that most of the E. coli isolated from chicken in this study belonged to group B1 (36.0%),group D (28.0%), group A (27.2%) and group B2 (8.8%). Multiplex PCR analysis demonstrated that 96 (78.6%) of the E. coli isolates harbored at least one virulencegene, while 29 (23.3%) did not contain any virulence genes tested. The most prevalent virulence genes identified were iss (51.2%), followed by iucD (36.0%),tsh (32.8%), vat (16.0%), astA (13.6%), irp2 (11.2%), papC (9.6%) and the least is cva/cvi gene (0%). None of the isolates harbored more than four virulence genes.Each of phylogenetic groups presented with different combinations of virulence genes, with no specific combinations of virulence genes found to correlate withE. coli phylogroups. None of the E. coli isolates harbored more than four virulence genes, suggesting that E. coli isolates from chicken in this study appear to bederived from commensal strains and may relate to environmental predispose factors especially stress factors in the host to establish infection.
Show more [+] Less [-]Fascioliasis in an adult draught buffalo in Malaysia: a case report
2017
Rosilawati K. | Ramli S. | Saipul Bahari A. R.
Fascioliasis is an importantparasitic disease caused by the liver flukeFasciola gigantica in Malaysia. Theinfestation of liver fluke in ruminants;cattle, sheep, goats and buffaloes can resultin economic losses to the country mainlydue to the drop in livestock production,reduction in growth rate, condemnationof liver, reduction in draught power andhigh usage of anthelmintics. This paperdescribes a case of liver fluke infestationin a two year old male buffalo that waseventually slaughtered. The buffalowas reported to be emaciated and wasreared in an oil palm plantation for useas draught power in buffalo-drawn cartsloaded with oil palm bunches; alongwith 18 other buffaloes of various ages.Previously, there were two cases of buffalodeaths from the same herd. The organand faecal samples were sent to KuantanRegional Veterinary Laboratory (RVL)for diagnostic work up. The receivedsamples were then sent to parasitology,histopathology and bacteriology sectionsfor laboratory analysis and confirmationon the pathogens. The bacteriology resultshowed Escherichia coli isolated in allinternal organs. Presence of adult flukes(Fasciola gigantica) were observed inthe bile duct of the liver through grossexamination and also histopathologicalevaluation and supported by the positiveresult of Fasciola ova via sedimentationtest conducted from the faecal sample, thusjustifying the final diagnosis as severe liverfluke infestation leading to emaciation andsimultaneously having colibacillosis.
Show more [+] Less [-]Antibiotic resistance of Escherichia coli isolated from chicken in Malaysia
2016
Ramlan M. | Khairani-Bejo S. | Khoo, E. | Roseliza R. | Zunita Z. | Rosnah Y.
Colibacillosis is animportant disease affecting the poultryindustry in many countries, caused bythe Avian Pathogenic E. coli (APEC):it manifests as various clinical signs. Itcontributes significantly to economicloss for poultry farmers as a result ofhigh mortality and morbidity in poultry.To overcome this, antibiotics have beenwidely used to eliminate E. coli infectionin poultry farms in recent years. Treatmentwith antibiotics has been considered as avital regimen to control E. coli infectionat the farm level for many years. However,high frequency of antibiotic resistance ofE. coli isolates from chicken has becomethe centre of attention due to public healthimportance. The aim of the present study isto determine the multidrug resistant profilesof E. coli strains isolated from chicken.E. coli isolates obtained from clinicalcases were re-identified and classified byconventional methods. Multidrug resistantprofiles against 13 different antibiotics of125 E. coli isolates were determined byusing disk diffusion method accordingto Clinical Laboratory Standard Institute(CLSI). Antibiogram revealed that 81.6%of the E. coli isolates showed multidrugresistant profiles to different antibiotics.Most of the E.coli isolates were highlyresistant to erythromycin (52.8%), followedwith tetracycline (52.0%), spectinomycin(39.2%), trimethoprim (38.4%) andflumequin (37.6%). Out of 125 isolatestested, 19.2% were resistant to more thaneight antibiotics, with one isolates found tobe multidrug resistant to most of antibioticsexcept polymyxin B. These findings alsodemonstrated that most of the isolateswere susceptible to antibiotics commonlyused for E.coli infections treatment inpoultry with lowest resistant score againstpolymyxin B (92.8%) and colistin (92.0%).Moderate resistant profiles were observedtowards amoxycilin (25.6%), apramycin(16%), kanamycin (8.8%) and streptomycin(8.0%). High percentage of multidrugresistance was found among the E. coliisolated from chicken as an indicator tomore serious problems in animal health.Therefore, continuous surveillance of antibiotic resistance profiles in chicken andother food animals is crucial to ensure foodchain safety
Show more [+] Less [-]Virulence gene profiles of avian pathogenic <i>Escherichia coli</i> isolated from chickens with colibacillosis in Bulawayo, Zimbabwe
2015
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.
Show more [+] Less [-]Investigation of high fatality among lambs in sheep farms in Pekan, Pahang, Malaysia
2016
Rosilawati K. | Nurul Faizah Z. | Saipul Bahari A. R.
This study was conducted to investigate the high mortality of young lambs in two sheep farms in Pekan, Pahang over a period of 3 years. Samples from postmortem of 1,451 lambs below one year of age by a farm veterinarian were submitted for laboratory diagnosis at the Bacteriology Section of the Regional Veterinary Diagnostic Laboratory in Kuantan. Escherichia coli is the most commonly recorded bacteria with 161 lambs diagnosed in 2013. In 2014 and 2015, there was a decrease in occurrence of E. coli related deaths, with 120 and 75lambs respectively. A total of 25% of the cases showed Escherichia coli positive by culture on blood agar and MacConkey agar, and confirmed by biochemical tests. A total of 21% of the cases were positive for staphylococcus sp, 3% and 6% for Streptococcus sp and Klebsiella pneumonia, respectively. Other bacteria were isolated in 45% of the cases. It was further noted that a total of 285 lambs between the ages of one to four months of age followed by 58 lambs (20%) less than one month old had E.coli isolation. It is also noteworthy that there were 10 lambs with E.coli infection in one to fourteen day-old lambs during the 3-year period from January 2013 to December 2015. This information was collated as a result of routine diagnosis of field cases submitted and with the intention of highlighting the common pathogens causing high mortality in local small ruminant farms so that preventive action may be taken for future farming ventures. E. coli infections or Colibacillosis is an important finding and indicator of poor management including poor nutrition, hygiene and environmental contamination which can reduce animal immunity and render it susceptible to other infections.
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