57 research outputs found
Performance analysis of anaplasma antibody competitive ELISA using the ROC curve for screening of anaplasmosis in camel populations in Egypt
Anaplasmosis is a tick-born and potential zoonotic disease caused by Anaplasma (A.) phagocytophilum, A. ovis, A. platys and A. capra. Anaplasma marginale affecting bovines and camels causing significant economic losses. Camels as an integral part of the socio-economic lifestyle of nomads in semi-arid to arid ecosystems are prone to suffer from subclinical Anaplasma infections. This study aimed to determine the performance and adaptation of commercial competitive Anaplasma ELISA (cELISA) as a tool for screening the seroprevalence of anaplasmosis whitin the camel populations in Egypt. This study was based on the serological investigation of 437 camel sera collected between 2015 and 2016 during a Q fever prevalence study in Egypt using commercially available cELISA for the detection of antibodies specific for Anaplasma in bovine serum. The receiver operating characteristic (ROC) curve, an analysis method for optimizing cutoff values in cELISAs, was used to estimate the sensitivity and specificity using 76 true as serological positive (n = 7) and negative (n = 60) for Anaplasma antibodies. ROC curve analysis was done for 7 true positive and 60 true negative bovine samples and 7 true positive and 29 true negative camel samples serum. Real time PCR and/or conventional PCR was applied to confirm Anaplasma spp. specific-DNA in camel serum as an indication of a true positive and true negative for ROC analysis. Chi square analysis was performed to estimate the association between risk factors and anaplasmosis in camels. The cutoff value was determined as 0.42 (p value ≤ 0.001). Data simulation with randomly generated values revealed a cutoff value of 0.417 (p ≤ 0.001) with resulting 58.1% Se and 97.8% Sp. Seven true positive and 29 true negative camel serum samples was confirmed by PCR. Using the estimated cut off, the seroprevalence in the Nile Valley and Delta and the Eastern Desert domain was 47.4% and 46.4%, respectively. The potential risk factors as domains and origin of animals were less significantly associated with the prevalence of anaplasmosis (domains: χ(2) = 41.8, p value ≤ 0.001 and origin: χ(2) = 42.56, p value ≤ 0.001). Raising awareness especially for veterinarians and animal owners will significantly contribute to the best understanding of anaplasmosis in camels in Egypt. Alternative (in silico) validation techniques and preliminary prevalence studies are mandatory towards the control of neglected anaplasmosis in the camel population
Seroprevalence and Molecular Detection of Bovine Anaplasmosis in Egypt
Bovine anaplasmosis is a tick-borne disease with zoonotic potential, caused by the obligate intracellular bacterium Anaplasma marginale. The disease is distributed worldwide in tropical and subtropical regions. The economic losses from anaplasmosis in animals is of significant importance because it causes severe morbidity and mortality in cattle. Recovered animals may become persistent carriers. Epidemiological information on the actual status of bovine anaplasmosis in Egypt is scarce. Thus, this study aimed to determine anti-Anaplasma antibody and DNA in serum samples using ELISA and PCR, respectively. In total, 758 bovine sera were collected from cattle farms located in 24 Egyptian governorates in 2015 to 2016. Sera were analyzed with the commercially available ‘Anaplasma antibody competitive ELISA v2’ kit and ‘AmpliTest Anaplasma/Ehrlichia spp. real time TaqMan TM PCR. Anaplasma spp. antibodies were detected in 140 (18.5%) (CI: 15.8–21.4%) of the investigated sera by ELISA, and Anaplasma/Ehrlichia-DNA was detected in 40 (5.3%) (CI: 3.8–7.1%) of the positive sera by real time PCR. Co-detection of both Anaplasma spp. and Coxiella burnetii-specific antibodies was proven in 30 (4%) of the investigated sera. The results of this work confirm the significant prevalence of bovine anaplasmosis in Egypt. Raising awareness in decision makers of the public health, veterinarians and animal owners is required to reduce the spread of infection
Characterization of Methicillin-Resistant Staphylococcus aureus Isolated from Healthy Turkeys and Broilers Using DNA Microarrays
Methicillin-resistant Staphylococcus aureus (MRSA) is a major human health
problem and recently, domestic animals are described as carriers and possible
reservoirs. Twenty seven S. aureus isolates from five turkey farms (n = 18)
and two broiler farms (n = 9) were obtained by culturing of choana and skin
swabs from apparently healthy birds, identified by Taqman-based real-time
duplex nuc-mecA-PCR and characterized by spa typing as well as by a DNA
microarray based assay which covered, amongst others, a considerable number of
antibiotic resistance genes, species controls, and virulence markers. The
antimicrobial susceptibility profiles were tested by agar diffusion assays and
genotypically confirmed by the microarray. Five different spa types (3 in
turkeys and 2 in broilers) were detected. The majority of MRSA isolates
(24/27) belonged to clonal complex 398-MRSA-V. The most frequently occurring
spa types were accordingly t011, t034, and t899. A single CC5-MRSA-III
isolated from turkey and CC398-MRSA with an unidentified/truncated SCCmec
element in turkey and broiler were additionally detected. The phenotypic
antimicrobial resistance profiles of S. aureus isolated from both turkeys and
broilers against 14 different antimicrobials showed that all isolates were
resistant to ampicillin, cefoxitin, oxacillin, doxycycline, and tetracycline.
Moreover, all S. aureus isolated from broilers were resistant to erythromycin
and azithromycin. All isolates were susceptible to gentamicin,
chloramphenicol, sulphonamides, and fusidic acid. The resistance rate against
ciprofloxacin was 55.6% in broiler isolates and 42.1% in turkey isolates. All
tetracycline resistant isolates possessed genes tetK/M. All erythromycin-
resistant broiler isolates carried ermA. Only one broiler isolate (11.1%)
carried genes ermA, ermB, and ermC, while 55.6% of turkey isolates possessed
ermA and ermB genes. Neither PVL genes (lukF/S-PV), animal-associated
leukocidin (lukM and luk-P83) nor the gene encoding the toxic shock syndrome
toxin (tst1) were found in turkey and broiler isolates. In conclusion, the
detection of MRSA in healthy turkeys and broilers with even additional
antibiotic resistance markers is of major public health concern. The
difference in antibiotic resistance and virulence markers between MRSA
isolates from turkeys and broilers was addressed
Occurrence of Salmonella enterica and Escherichia coli in raw chicken and beef meat in northern Egypt and dissemination of their antibiotic resistance markers
Background The global incidence of foodborne infections and antibiotic
resistance is recently increased and considered of public health concern.
Currently, scarcely information is available on foodborne infections and ESBL
associated with poultry and beef meat in Egypt. Methods In total, 180 chicken
and beef meat samples as well as internal organs were collected from different
districts in northern Egypt. The samples were investigated for the prevalence
and antibiotic resistance of Salmonella enterica serovars and Escherichia
coli. All isolates were investigated for harbouring class 1 and class 2
integrons. Results Out of 180 investigated samples 15 S. enterica (8.3%) and
21 E. coli (11.7%) were isolated and identified. S. enterica isolates were
typed as 9 S. Typhimurium (60.0%), 3 S. Paratyphi A (20.0%), 2 S. Enteritidis
(13.3%) and 1 S. Kentucky (6.7%). Twenty-one E. coli isolates were serotyped
into O1, O18, O20, O78, O103, O119, O126, O145, O146 and O158. The phenotypic
antibiotic resistance profiles of S. enterica serovars to ampicillin,
cefotaxime, cefpodoxime, trimethoprim/sulphamethoxazole and tetracycline were
86.7, 80.0, 60.0, 53.3 and 40.0%, respectively. Isolated E. coli were
resistant to tetracycline (80.9%), ampicillin (71.4%), streptomycin,
trimethoprim/sulphamethoxazole (61.9% for each) and cefotaxime (33.3%). The
dissemination of genes coding for ESBL and AmpC β-lactamase in S. enterica
isolates included bla CTX-M (73.3%), bla TEM (73.3%) and bla CMY (13.3%). In
E. coli isolates bla TEM, bla CTX-M and bla OXA were identified in 52.4, 42.9
and 14.3%, respectively. The plasmid-mediated quinolone resistance genes
identified in S. enterica were qnrA (33.3%), qnrB (20.0%) and qnrS (6.7%)
while qnrA and qnrB were detected in 33.3% of E. coli isolates. Class 1
integron was detected in 13.3% of S. enterica and in 14.3% of E. coli
isolates. Class 2 integron as well as the colistin resistance gene mcr-1 was
not found in any of E. coli or S. enterica isolates. Conclusions This study
showed high prevalence of S. enterica and E. coli as foodborne pathogens in
raw chicken and beef meat in Nile Delta, Egypt. The emergence of antimicrobial
resistance in S. enterica and E. coli isolates is of public health concern in
Egypt. Molecular biological investigation elucidated the presence of genes
associated with antibiotic resistance as well as class 1 integron in S.
enterica and E. coli
Serological and Molecular Identification of Brucella spp. in Pigs from Cairo and Giza Governorates, Egypt
Brucellosis is considered as endemic disease of animals and humans since thousands of years in Egypt. However, brucellosis in pigs has never been reported in Egypt. Thus, serological and molecular assays were applied to detect anti-Brucella antibodies and DNA in serum samples collected from pigs. In total 331 blood samples collected from male and female pigs at slaughterhouses of Cairo and Giza governorates were investigated using Brucella c- and i-ELISA and Brucella real-time PCR. Anti-Brucella antibodies were detected in 16 (4.83%) and 36 (10.8%) sera by i-ELISA and c-ELISA, respectively. Brucella DNA was detected in 10 (3.02%) seropositive samples and identified as Brucella melitensis (7/10) and Brucella suis (3/10). A higher prevelance was found in boars. This is the first study investigating pig brucellosis in Egypt. The results of this study will raise awareness for brucellosis in these farm animals and will help to develop effective control strategies
Prevalence, genotyping and risk factors of thermophilic Campylobacter spreading in organic turkey farms in Germany
Background The need for organic food of animal origin has increased rapidly in
recent years. However, effects of organic animal husbandry on food safety have
not been rigorously tested especially in meat turkey flocks. This study
provides for the first time an overview on the prevalence and genetic
diversity of Campylobacter species (spp.) in five organic meat turkey farms
located in different regions in Germany, as well as on potential risk factors
of bacterial spreading. Thirty cloacal swabs as well as water samples and
darkling beetles were collected from each flock and examined for the presence
of Campylobacter by conventional and molecular biological methods. The
isolates were genotyped by flaA-RFLP. Results Campylobacter spp. were detected
in cloacal swabs in all 5 turkey flocks with prevalence ranged from 90.0 to
100 %. 13 cloacal swabs collected from birds in farm III and IV were harboured
mixed population of thermophilic campylobacters. In total, from 158
Campylobacter isolated from turkeys 89 (56.33 %) were identified as C. coli
and 69 (43.76 %) as C. jejuni. Three Campylobacter (2 C. jejuni and 1 C. coli)
were detected in drinkers of two farms and 3 C. coli were isolated from
darkling beetles of one farm. No Campylobacter were isolated from main water
tanks. flaA-RFLP assay showed that turkey farms can harbour more than one
genotype. In a single turkey two different genotypes could be detected. The
genotypes of campylobacters isolated from water samples or beetles were
identical with those isolated from turkeys. No effect was found of some
environmental parameters [ammonia concentration (NH3), carbon dioxide
concentration (CO2), relative humidity (RH) and air temperature)] on
Campylobacter prevalence in organic turkey farms. Additionally, drinking water
and darkling beetles might be considered as risk factors for the spreading of
Campylobacter in turkey flocks. Conclusions This study highlights the high
prevalence and genotypic diversity of Campylobacter spp. isolated from organic
turkey flocks. Further research is needed to assess other potential risk
factors responsible for bacteria spreading in order to mitigate the spread of
Campylobacter in organic turkey flocks by improving biosecurity control
measures
Identification, Genotyping and Antimicrobial Susceptibility Testing of Brucella spp. Isolated from Livestock in Egypt
Brucellosis is a highly contagious zoonosis worldwide with economic and public health impacts. The aim of the present study was to identify Brucella (B.) spp. isolated from animal populations located in different districts of Egypt and to determine their antimicrobial resistance. In total, 34-suspected Brucella isolates were recovered from lymph nodes, milk, and fetal abomasal contents of infected cattle, buffaloes, sheep, and goats from nine districts in Egypt. The isolates were identified by microbiological methods and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Differentiation and genotyping were confirmed using multiplex PCR for B. abortus, Brucella melitensis, Brucella ovis, and Brucella suis (AMOS) and Bruce-ladder PCR. Antimicrobial susceptibility testing against clinically used antimicrobial agents (chloramphenicol, ciprofloxacin, erythromycin, gentamicin, imipenem, rifampicin, streptomycin, and tetracycline) was performed using E-Test. The antimicrobial resistance-associated genes and mutations in Brucella isolates were confirmed using molecular tools. In total, 29 Brucella isolates (eight B. abortus biovar 1 and 21 B. melitensis biovar 3) were identified and typed. The resistance of B. melitensis to ciprofloxacin, erythromycin, imipenem, rifampicin, and streptomycin were 76.2%, 19.0%, 76.2%, 66.7%, and 4.8%, respectively. Whereas, 25.0%, 87.5%, 25.0%, and 37.5% of B. abortus were resistant to ciprofloxacin, erythromycin, imipenem, and rifampicin, respectively. Mutations in the rpoB gene associated with rifampicin resistance were identified in all phenotypically resistant isolates. Mutations in gyrA and gyrB genes associated with ciprofloxacin resistance were identified in four phenotypically resistant isolates of B. melitensis. This is the first study highlighting the antimicrobial resistance in Brucella isolated from different animal species in Egypt. Mutations detected in genes associated with antimicrobial resistance unravel the molecular mechanisms of resistance in Brucella isolates from Egypt. The mutations in the rpoB gene in phenotypically resistant B. abortus isolates in this study were reported for the first time in Egypt
Evolution of Antibiotic Resistance of Coagulase-Negative Staphylococci Isolated from Healthy Turkeys in Egypt: First Report of Linezolid Resistance
Coagulase-negative staphylococci (CoNS) are gaining much attention as causative agents of serious nosocomial infections in humans. This study aimed to determine the prevalence and phenotypic antimicrobial resistance of CoNS as well as the presence of resistance-associated genes in CoNS isolated from turkey farms in Egypt. Two hundred and fifty cloacal swabs were collected from apparently healthy turkeys in Egypt. Suspected isolates were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The susceptibility testing of CoNS isolates against 20 antimicrobial agents was performed using the broth microdilution test. The presence of resistance-associated genes like mecA, vanA, blaZ, erm(A), erm(B), erm(C), aac-aphD, optrA, valS, and cfr was determined. Thirty-nine CoNS were identified. All isolates were phenotypically resistant to trimethoprim/sulfamethoxazole, penicillin, ampicillin, and tetracycline. The resistance rates to erythromycin, chloramphenicol, oxacillin, daptomycin, and tigecycline were 97.4%, 94.9%, 92.3%, 89.7%, and 87.2%, respectively. Thirty-one isolates were resistant to linezolid (79.5%). Low resistance rate was detected for both imipenem and vancomycin (12.8%). The erm(C) gene was identified in all erythromycin phenotypically resistant isolates, whereas two resistant isolates possessed three resistance-conferring genes erm(A), erm(B), and erm(C). The cfr and optrA genes were detected in 11 (35.5%) and 12 (38.7%) of the 31 linezolid-resistant isolates. The mecA, aac-aphD, and blaZ genes were identified in 22.2%, 41.9%, and 2.6% of phenotypically resistant isolates to oxacillin, gentamicin, and penicillin, respectively. This is the first study revealing the correlation between linezolid resistance and presence of cfr and optrA genes in CoNS isolates from Egypt, and it can help to improve knowledge about the linezolid resistance mechanism
Genomic insight into Campylobacter jejuni isolated from commercial turkey flocks in Germany using whole-genome sequencing analysis
Campylobacter (C.) jejuni is a zoonotic bacterium of public health significance. The present investigation was designed to assess the epidemiology and genetic heterogeneity of C. jejuni recovered from commercial turkey farms in Germany using whole-genome sequencing. The Illumina MiSeq® technology was used to sequence 66 C. jejuni isolates obtained between 2010 and 2011 from commercial meat turkey flocks located in ten German federal states. Phenotypic antimicrobial resistance was determined. Phylogeny, resistome, plasmidome and virulome profiles were analyzed using whole-genome sequencing data. Genetic resistance markers were identified with bioinformatics tools (AMRFinder, ResFinder, NCBI and ABRicate) and compared with the phenotypic antimicrobial resistance. The isolates were assigned to 28 different sequence types and 11 clonal complexes. The average pairwise single nucleotide-polymorphisms distance of 14,585 SNPs (range: 0–26,540 SNPs) revealed a high genetic distinction between the isolates. Thirteen virulence-associated genes were identified in C. jejuni isolates. Most of the isolates harbored the genes flaA (83.3%) and flaB (78.8%). The wlaN gene associated with the Guillain–Barré syndrome was detected in nine (13.6%) isolates. The genes for resistance to ampicillin (blaOXA), tetracycline [tet(O)], neomycin [aph(3')-IIIa], streptomycin (aadE) and streptothricin (sat4) were detected in isolated C. jejuni using WGS. A gene cluster comprising the genes sat4, aph(3′)-IIIa and aadE was present in six isolates. The single point mutation T86I in the housekeeping gene gyrA conferring resistance to quinolones was retrieved in 93.6% of phenotypically fluoroquinolone-resistant isolates. Five phenotypically erythromycin-susceptible isolates carried the mutation A103V in the gene for the ribosomal protein L22 inferring macrolide resistance. An assortment of 13 β-lactam resistance genes (blaOXA variants) was detected in 58 C. jejuni isolates. Out of 66 sequenced isolates, 28 (42.4%) carried plasmid-borne contigs. Six isolates harbored a pTet-like plasmid-borne contig which carries the tet(O) gene. This study emphasized the potential of whole-genome sequencing to ameliorate the routine surveillance of C. jejuni. Whole-genome sequencing can predict antimicrobial resistance with a high degree of accuracy. However, resistance gene databases need curation and updates to revoke inaccuracy when using WGS-based analysis pipelines for AMR detection
Epidemiological, molecular characterization and antibiotic resistance of Salmonella enterica serovars isolated from chicken farms in Egypt
Background Salmonella is one of major causes of foodborne outbreaks globally.
This study was conducted to estimate the prevalence, typing and antibiotic
susceptibilities of Salmonella enterica serovars isolated from 41 broiler
chicken farms located in Kafr El-Sheikh Province in Northern Egypt during
2014–2015. The clinical signs and mortalities were observed. Results In total
615 clinical samples were collected from broiler flocks from different organs
(liver, intestinal content and gall bladder). Salmonella infection was
identified in 17 (41%) broiler chicken flocks and 67 Salmonella isolates were
collected. Recovered isolates were serotyped as 58 (86.6%) S. enterica serovar
Typhimurium, 6 (9%) S. enterica serovar Enteritidis and 3 (4.5%) were non-
typable. The significant high mortality rate was observed only in 1-week-old
chicks. sopE gene was detected in 92.5% of the isolates which indicating their
ability to infect humans. All S. enterica serovar Enteritidis isolates were
susceptible to all tested antimicrobials. The phenotypically resistant S.
enterica serovar Typhimurium isolates against ampicillin, tetracycline,
sulphamethoxazole and chloramphenicol were harbouring BlaTEM, (tetA and tetC),
(sul1 and sul3) and (cat1 and floR), respectively. The sensitivity rate of S.
enterica serovar Typhimurium to gentamycin, trimethoprim/sulphamethoxazole and
streptomycin were 100, 94.8, 89.7%, respectively. The silent streptomycin
antimicrobial cassettes were detected in all Salmonella serovars. A class one
integron (dfrA12, orfF and aadA2) was identified in three of S. enterica
serovar Typhimurium strains. Conclusions To the best of our knowledge, this
study considered first report discussing the prevalence, genotyping,
antibiotic susceptibility and public health significance of S. enterica
serovars in broilers farms of different ages in Delta Egypt. Further studies
are mandatory to verify the location of some resistance genes that are within
or associated with the class one integron
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