1.The detection of adherence factorsby Escherichia coli cause of urinary tract infectionsin Ulaanbaatar, Mongolia
Munkhdelger Ya ; Davaasuren S ; Dolgorjav B ; Gerelee A ; Oyunchimeg R ; Sarantuya J
Mongolian Medical Sciences 2014;168(2):5-8
INTRODUCTION:
Urinary tract infections among the most common bacterial infectious diseases encountered at all
ages. Escherichia coli are being the etiologic agent in 50–80%. Therefore, it is an important public
health problem. E.coli causing urinary tract infections express pilli, fimbriae and others adherence
virulence factors.
GOAL:
To detect the some adherence virulence factors of Uropathogenic Escherichia coli (UPEC) in
Ulaanbaatar, Mongolia
MATERIALS AND METHODS:
A total of 76E.colisampleswere collected. These samples were positive bacteriological examination
of urine, performed at the bacteriological laboratory of the State Central Third Hospital and State
Central First Hospital, Ulaanbaatar, Mongolia. The biofilm formation was evaluated by the growth
rate of E.coli on plastic surface.The detection of the virulence factors type 1 fimbriae (fimA gene) and
P-fimbriae (papC) was performed by multiplex PCR using gene specific primers.Curli expression
was determined by using congo red agar.
RESULTS:
The evaluation of bacterial biofilm formation using 96 well plates showed 40 negative (52.6%),
32 weak biofilm (42.1%) and 4 moderate biofilm (5.3%) formation for E.coli and no strong biofilm
forming strain was detected. The cell surface protein (curli) was detected by Congo red agar. The
result was 71% positive for studied E.coli strains. The detection result of pili genes by multiplex
PCR showed that fimH gene detected for 73 (96.1%) and papC gene detected for 18 (23.7%) E.coli
cultures.
CONCLUSION: Almost half of surveyed Uropathogenic E.coli isolated in Ulaanbaatar, Mongolia had
ability of biofilm formation and it has been determined by the bacterial surface protein (curli), which
is one of bacterial adherence factors, may cause biofilm formation.
2.Relation between adherence factors and the phylogenetic group of extraintestinal pathogenic Escherichia coli
Munkhdelger Ya ; Nyamaa G ; Undarmaa G ; Oyunchimeg R ; Munguntuul T ; Altantsetseg D ; Munkh-Od Ts ; Sarantuya J
Innovation 2015;9(2):28-31
Extraintestinal pathogenic Escherichia coli (ExPEC), the specialized strains ofE.coli that cause most extraintestinal infections, represent a major but littleappreciated health threat. Phylogenetic analysis has shown that ExPEC is composedof four main phylogenetic groups (A,B1, B2, and D) and that virulent extraintestinalstrains mainly belong to groups B2 and D.In this study, we aimed to assess therelation between adherence virulence and phylogenetic groups of ExPEC.A total of 161 E.coli samples were collected. Out of these 17 (10.6%) werefrom pus, 66 (41 %) from urine, 78 (48.4%) from cervical swab. The phylogeneticgroups and 6 virulence genes (fimH, papC, papGII, papGIII, fa/draBC,andSfa/focDE) encoding adhesins were identified by triplex PCR. Phylogeneticgroups distribution was as follows: B1 10.5%, A 24.7%, B2 25.3%, and D 38.9%. Virulence genes prevalence was fimH 90.1%, papC 23%, papGII 16.8%, papGIII1.9%, Afa/draBC 11.8%, andSfa/focDE 5.6%. The cell surface protein (curli) wasdetected 50,3% by Congo red agar. In conclusion: The most isolated strainsbelonged to the phylogenetic group B2 and D. The phylogenetic groups weresignificantly associated with some genes encoding
adhesins (fimH, papC) and cellsurface protein (curli).
3.Detection of virulent of Klebsiella isolates from ñlinical samples
Undarmaa G ; Munkhdelger YA ; Bayarlakh B ; Oyunchimeg R ; Dolgorjav B ; Sarantuya J
Mongolian Medical Sciences 2016;175(1):21-25
IntroductionKlebsiella spp is a well-known opportunistic pathogen associated with nosocomial infections such asurinary tract, septicaemia and pneumonia number of multi-drug resistant strains and infections causedby Klebsiella has progressively increased, causing treatment limitations.GoalIdentify of phenotype of Klebseilla isolates from ñlinical samplesMaterials and MethodsA total of 112 Klebsiella strains were isolated from clinical samples in State Central First Hospital and StateCentral Third Hospital from July 2015 through December 2015. The bacterial isolates were identifi edaccording to cultural characteristics, biochemical test and API20E. The serum resistance, capsule andhypermucoviscosity, cell surface protein (curly), a-hemolysin and ability to form biofi lm were sought byphenotypic assays. Antimicrobial susceptibility was tested by diffusion method.ResultA total of 112 Klebsiella samples were collected. The bacterial isolates were identifi ed according tocultural characteristics, biochemical test and API20E, the results revealed that 16.1 percent isolateswere identifi ed as K.oxytoca all of them 83.9 percent isolates were belong to K.pneumonia. Therewere observed for ampicillin (99 percent), nitrofurantoin (53.6 percent), cepalotin (50.6 percent) and51 percent of isolates were considered as a multiple drug resistant. Serum resistance properties ofK.pneumoniae was resistance 89.4 percent, intermediately susceptible 4.3 percent, sensitive 6.4percent and for K.oxytoca resistance 88.9 percent, intermediately susceptible 5.6 percent, sensitive 5.6percent. The hemolysin àalpha was detected in 32.2 percent, and gamma, beta in 66.96 percent, 0.9percent respectively. The capsule was observed in 46.5 percent and hypermucoviscosity in 27.7 percentof isolates. The cell surface protein (curly) and biofi lm were detected in 100 percent.Conclusion:Both K.pneumoniae and K.oxytoca isolates from clinical samples have similar virulent properties, andthe a-hemolysin and hypermucoviscosity positive isolates were more resistance to antibiotics.
4. Relation between adherence factors and the phylogenetic group of extraintestinal pathogenic Escherichia coli
Munkhdelger YA ; Nyamaa G ; Undarmaa G ; Oyunchimeg R ; Munguntuul T ; Altantsetseg D ; Munkh-Od TS ; Sarantuya J
Innovation 2015;9(2):28-31
Extraintestinal pathogenic Escherichia coli (ExPEC), the specialized strains ofE.coli that cause most extraintestinal infections, represent a major but littleappreciated health threat. Phylogenetic analysis has shown that ExPEC is composedof four main phylogenetic groups (A,B1, B2, and D) and that virulent extraintestinalstrains mainly belong to groups B2 and D.In this study, we aimed to assess therelation between adherence virulence and phylogenetic groups of ExPEC.A total of 161 E.coli samples were collected. Out of these 17 (10.6%) werefrom pus, 66 (41 %) from urine, 78 (48.4%) from cervical swab. The phylogeneticgroups and 6 virulence genes (fimH, papC, papGII, papGIII, fa/draBC,andSfa/focDE) encoding adhesins were identified by triplex PCR. Phylogeneticgroups distribution was as follows: B1 10.5%, A 24.7%, B2 25.3%, and D 38.9%. Virulence genes prevalence was fimH 90.1%, papC 23%, papGII 16.8%, papGIII1.9%, Afa/draBC 11.8%, andSfa/focDE 5.6%. The cell surface protein (curli) wasdetected 50,3% by Congo red agar. In conclusion: The most isolated strainsbelonged to the phylogenetic group B2 and D. The phylogenetic groups weresignificantly associated with some genes encodingadhesins (fimH, papC) and cellsurface protein (curli).
5.Non-invasive markers for staging fibrosis in patients with chronic hepatitis Delta
Sarantuya G ; Sumiya D ; Selenge J ; Uranbailgal E ; Otgonbayar R ; Munkhbat B ; Bira N
Mongolian Medical Sciences 2021;195(1):18-24
Introduction:
Determining stages of liver fibrosis in chronic liver disease is essential for clinical practice such as
decision making on medical treatment, setting the interval of follow-up examination for its complication,
screening intervals for hepatocellular carcinoma.
Goal:
We compared non-invasive fibrosis markers among the patients with chronic hepatitis Delta.
Materials and Methods:
Totally 70 patients with chronic hepatitis D enrolled into this study. The blood samples were examined
for complete blood count, liver function test and serum M2BPGi level. Non-invasive markers such
as AAR, APRI, Fib-4 scores were calculated. Those with AAR >1, APRI >0.7, FIB-4 >1.45 were
considered with advanced fibrosis. All patients underwent liver stiffness measurement using FibroScan
M2 probe. The cutoff values of FibroScan for advanced fibrosis were 9 kPa for patient with normal
transaminase level and 11 kPa for patients with elevated transaminase.
Results:
Advanced fibrosis was observed in 25.7%, 38.6% and 38.6% by AAR, APRI and Fib-4 score,
respectively. When cut-off levels of serum M2BPGi for advanced fibrosis was 2.2 COI, 35.7% had
advanced fibrosis. FibroScan tests showed 34.4% had advanced fibrosis. The AUROC of M2BPGi
were 0.894 and 0.827 for predicting advanced fibrosis and liver cirrhosis.
Conclusion
Serum M2BPGi and FibroScan would be reliable diagnostic tool for identifying liver
fibrosis in Mongolian patients with chronic hepatitis D.
6.ISOLATION AND PURIFICATION OF IMMUNE MODULATING LACTOFERRIN FROM MONGOL BOVINE COLOSTRUM
Chingunjav E ; Jambal B ; Amarsaikhan B ; Gerelmaa T ; Narantsetseg L ; Sarantuya R ; Bilegtsaikhan Ts ; Purevjargal N ; Tengis A ; Javkhlan B ; Tsendmaa Ts ; Galindev B ; Munkhtulga L ; Nyambayar D ; Munkhbat B ; Baigalmaa B
Innovation 2017;11(1):30-33
BACKGROUND
Bovine colostrums is the milk secreted by cows during the first few days after parturition. It
contains many essential nutrients and bioactive components, including growth factors,
immunoglobulins, lactoperoxidase, lactoferrin and cytokines ets. Lactoferrin has been reported
for its multifunctional properties such as antifungal, antibacterial, antiviral antioxidant and
anticancer activities. The aims of this study focused on the isolation and purification of lactoferrin
from Mongolian bovine colostrums. Lactoferrin purified using HiTrap DEAE an ion exchange
chromatography. Lactoferrin purification efficiency was about 60.5%. The single band of purified
lactoferrin has been observed in SDS-PAGE electrophoresis.
METHODS
Bovine colostrum was collected at a cow farm in the Darkhan province of Mongolia. At first
the cream was separated by centrifugation (10000 xg 20 min at 4oC). In order to separate the
whey, the samples were precipitated with 1mol/l to pH 4.6 and centrifuged at 10000 g 20 min
again. The samples of whey were stored at -18oC to the analysis. Lactoferrin was purified by
HiTrap DEAE an ion exchange chromatography using 0.005 M phosphate buffer (pH 7.7) and
linear gradient NaCl from 0.25M, 0.5M, 1M. During chromatography, protein in the eluents was
monitored by ultraviolet absorbation at 280 nm with the instrument. Purity test done by using
polyacrylamide gel electrophoresis under denaturated condition (SDS-PAGE) method by Laemmli
(1970). For HPLC determination of the lactoferrin by Shimadzu Nexera X2 HPLC system with UV/
VIS detector were used. Detection was carried out at the wavelength 280 nm. Separation was
performed on a chromatographic column Protein R C18 ,2.2 x 150 mm, 5 μm particle size. Linear
gradient and flow rate 0.2 ml/min were used. Mobile phase a consisted of water / acetonitrile/
trifluoroacetic acid ( 95:5:0.1). The column temperature was set at 40oC and injection volume
was 10 μl. Data were collected and evaluated by software Lab Solution. An external standard
method for quantification analytes was used.
RESULTS
Purified lactoferrin in the present study had a good concentration and purification efficiency
was about 60.5 %. Protein fraction from 1M NaCl gradient delivers sharp and clean peak to
HPLC chromatogram that fits intensity and retention time of standard bovine lactoferrin.
Ammount of lactoferrin in bovine colostrums was 0.6 mg/ml and it`s molecular weight 80 kDa as
a standard sample. The retention time of lactoferrin fraction which is purified by SDS-PAGE gel
electrophoresis. The peak of fraction same compared to the standard lactoferrin 5.8 minutes
by HPLC analysis.
CONCLUSION
Ion exchange chromatography shows reliable and easy isolation of lactoferrin from Mongol
bovine colostrum.