133 research outputs found
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
Effect of Boost Pressure on the In-Cylinder Tumble-Motion of GDI Engine under Steady-State Conditions using Stereoscopic-PIV
This paper experimentally investigates the effect of boost pressure on the in-cylinder flow field under steady-state conditions using stereoscopic particle image velocimetry (Stereo-PIV) through increasing the pressure difference across the intake valves. The FEV steady-state flow bench was modified to provide an optical access into the cylinder region. The stereoscopic PIV measurements were carried out at various pressure differences viz., 300, 450, and 600 mmH2O across the intake valves of Gasoline Direct Injection (GDI) head for the mid cylinder vertical tumble-plane. Ensemble average velocity vectors were used to characterize the tumble flow structure and for the calculation of tumble ratio and average turbulent kinetic energy. Moreover, the Proper Orthogonal Decomposition (POD) technique was conducted on the PIV measured velocity vector maps to identify the most energetic structures generated at different valve lifts and pressure differences. The results of stereoscopic PIV measurements showed that the overall in-cylinder flow structures were mainly dependent on the valve lift irrespective of the applied pressure difference. However, the level of the turbulence kinetic energy increased as the boost pressure increased
Effect of boost pressure on the in-cylinder tumble- motion of GDI engine under steady-state conditions using Stereoscopic-PIV
This paper experimentally investigates the effect of boost pressure on the in-cylinder flow field under steady-state conditions using stereoscopic particle image velocimetry (Stereo-PIV) through increasing the pressure difference across the intake valves. The FEV steady-state flow bench was modified to provide an optical access into the cylinder region. The stereoscopic PIV measurements were carried out at various pressure differences viz., 300, 450, and 600 mmH2O across the intake valves of Gasoline Direct Injection (GDI) head for the mid cylinder vertical tumble-plane. Ensemble average velocity vectors were used to characterize the tumble flow structure and for the calculation of tumble ratio and average turbulent kinetic energy. Moreover, the Proper Orthogonal Decomposition (POD) technique was conducted on the PIV measured velocity vector maps to identify the most energetic structures generated at different valve lifts and pressure differences. The results of stereoscopic PIV measurements showed that the overall in-cylinder flow structures were mainly dependent on the valve lift irrespective of the applied pressure difference. However, the level of the turbulence kinetic energy increased as the boost pressure increased
Antimicrobial resistance of Campylobacter isolates from small scale and backyard chicken in Kenya
Background Thermophilic Campylobacter species are a major cause of bacterial
foodborne diarrhoea in humans worldwide. Poultry and their products are the
predominant source for human campylobacteriosis. Resistance of Campylobacter
to antibiotics is increasing worldwide, but little is known about the
antibiotic resistance in Campylobacter isolated from chicken in Kenya. In this
study, 35 suspected Campylobacter strains isolated from faeces and cloacal
swabs of chicken were tested for their susceptibility to seven antibiotics
using a broth microdilution assay and molecular biological investigations.
Results Overall, DNA of thermophilic Campylobacter was identified in 53
samples by PCR (34 C. jejuni, 18 C. coli and one mix of both species) but only
35 Campylobacter isolates (31 C. jejuni and 4 C. coli) could be re-cultivated
after transportation to Germany. Isolates were tested for their susceptibility
to antibiotics using a broth microdilution assay. Additionally, molecular
biological detection of antibiotic resistance genes was carried out. C. jejuni
isolates showed a high rate of resistance to nalidixic acid, tetracycline and
ciprofloxacin of 77.4, 71.0 and 71.0 %, respectively. Low resistance (25.8 %)
was detected for gentamicin and chloramphenicol. Multidrug resistance in C.
jejuni could be detected in 19 (61.3 %) isolates. Resistance pattern of C.
coli isolates was comparable. Resistance to ciprofloxacin was confirmed by
MAMA–PCR and PCR–RFLP in all phenotypically resistant isolates. The tet(O)
gene was detected only in 54.5 % of tetracycline resistant C. jejuni isolates.
The tet(A) gene, which is also responsible for tetracycline resistance, was
found in 90.3 % of C. jejuni and in all C. coli isolates. Thirteen
phenotypically erythromycin-resistant isolates could not be characterised by
using PCR–RFLP and MAMA–PCR. Conclusions To the best of our knowledge, this
study is the first report about resistance to antibiotics in thermophilic
Campylobacter originating from chicken in Kenya. Campylobacter spp. show a
high level of resistance to ciprofloxacin, nalidixic acid and tetracycline but
also a remarkable one to chloramphenicol and gentamicin and they are multidrug
resistant. Resistance to antibiotics is a global public health concern. In
Kenya, resistance surveillance needs further attention in the future. Efforts
to establish at least a National Laboratory with facilities for performing
phenotypic and genotypic characterization of thermophilic Campylobacter is
highly recommended
Seroprevalence and Molecular Identification of Brucella spp. in Camels in Egypt
Brucellosis is one of the most important worldwide zoonoses of many countries including Egypt. Camel brucellosis has not gained much attention in Egypt yet. This study is focused on the three governorates with the highest camel populations and the largest camel markets in the country to determine the disease seroprevalence and identify the Brucella species in local camel holdings. In total, 381 serum samples were collected from male and female camels from Giza, Aswan, and Al-Bahr Al-Ahmar (the Red Sea) governorates. Samples were serologically examined using the Rose–Bengal plate test (RBPT), indirect ELISA (i-ELISA), competitive ELISA (c-ELISA) and complement fixation test (CFT). Brucella antibodies were detected in 59 (15.5%), 87 (22.8%), 77 (20.2%) and 118 (31.0%) of sera by RBPT, i-ELISA, c-ELISA and CFT, respectively. Using real-time PCR, Brucella DNA was amplified in 32 (8.4%) seropositive samples including Brucella abortus (25/32), Brucella suis (5/32) and Brucella melitensis (2/32), defining a complex epidemiological status. To the best of our knowledge, this is the first study reporting Brucella suis DNA in camel serum. The risk-associated factors including age, sex, breed and geographical distribution were statistically analyzed, showing non-significant association with seroprevalence. The results of this study will raise awareness for camel brucellosis and help develop effective control strategies
Genotyping and antibiotic resistance of thermophilic Campylobacter isolated from chicken and pig meat in Vietnam
Background Campylobacter species are recognized as the most common cause of
foodborne bacterial gastroenteritis in humans. In this study nine
Campylobacter strains isolated from chicken meat and pork in Hanoi, Vietnam,
were characterized using molecular methods and tested for antibiotic
resistance. Results The nine isolates (eight C. jejuni and one C. coli) were
identified by multiplex PCR, and tested for the presence or absence of 29 gene
loci associated with virulence, lipooligosaccharide (LOS) biosynthesis and
further functions. flaA typing, multilocus sequence typing and microarray
assay investigation showed a high degree of genetic diversity among these
isolates. In all isolates motility genes (flaA, flaB, flhA, fliM),
colonization associated genes (cadF, docB), toxin production genes (cdtA,
cdtB, secD, secF), and the LOS biosynthesis gene pglB were detected. Eight
gene loci (fliY, virB11, Cje1278, Cj1434c, Cj1138, Cj1438c, Cj1440c, Cj1136)
could not be detected by PCR. A differing presence of the gene loci ciaB (22.2
%), Cje1280 (77.8 %), docC (66.7 %), and cgtB (55.6 %) was found. iamA, cdtC,
and the type 6 secretion system were present in all C. jejuni isolates but not
in C. coli. flaA typing resulted in five different genotypes within C. jejuni,
MLST classified the isolates into seven sequence types (ST-5155, ST-6736,
ST-2837, ST-4395, ST-5799, ST-4099 and ST-860). The microarray assay analysis
showed a high genetic diversity within Vietnamese Campylobacter isolates which
resulted in eight different types for C. jejuni. Antibiotic susceptibility
profiles showed that all isolates were sensitive to gentamicin and most
isolates (88.8 %) were sensitive to chloramphenicol, erythromycin and
streptomycin. Resistance rates to nalidixic acid, tetracycline and
ciprofloxacin were 88.9, 77.8 and 66.7 %, respectively. Conclusions To the
best of our knowledge, this study is the first report that shows high genetic
diversity and remarkable antibiotic resistance of Campylobacter strains
isolated from meat in Vietnam which can be considered of high public health
significance. These preliminary data show that large scale screenings are
justified to assess the relevance of Campylobacter infections on human health
in Vietnam
Characterization of Annona cherimola mill. Seed oil from Madeira Island: a possible biodiesel feedstock
The possibility of using Annona seed oil as an
added value product, namely as a source of biodiesel, is
explored. Milled Annona seeds were extracted with hexane
at room temperature (72 h) and at solvent boiling point
(6 h). Oil content was found to be 25 and 22.4% respec tively. The oil was characterized in terms of lipid compo sition (HPLC–APCI–MS and 13C NMR), resistance to
oxidation and acidity index. FAME composition was
determined by GC–MS and five major peaks were identi fied. Production of biodiesel from Annona’s seed oil was
achieved by base-catalyzed transesterification. Density,
viscosity, refraction coefficient, acid value, cold filter
plugging point, cloud point and oxidation stability were
measured. The iodine value and the ‘‘apparent cetane
number’’ were calculated. Density, viscosity, acid value,
iodine value, cold filter plugging point and cloud point
were within EN14214 specifications and the calculated
‘‘apparent cetane number’’ was also indicative of a suitable
product.info:eu-repo/semantics/publishedVersio
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