1.Identification of Enteroaggregative Escherichia coli in Mongolia
Munkhdelger Ya ; Undramaa G ; Munkh-Od Ts ; Nyamaa G ; Tsatsral E ; Sarantuya J
Mongolian Medical Sciences 2015;172(2):22-27
Introduction: Enteroaggregative Escherichia coli (EAEC) is an important agent of acute and
persistent diarrhea worldwide. Few cases have been reported in healthy children. EAEC strains are
characterized by aggregative adherence (AA) to HEp-2 cells, wherein bacteria are seen in “stacked
brick” aggregates attaching to HEp-2 cells and usually to the glass surface between cells.
Goal: To identify Enteroaggregative Escherihia coli using multiplex polymerase chain reaction (PCR)
and HEp-2 adherence assay in Ulaanbaatar, Mongolia
Materials and Methods:
A total of 329 E. coli strains were isolated from stool with diarrhea in National Center for Communicable
Diseases from July 2012 through September 2014. All specimens were processed by routine
microbiological and biochemical tests in the bacteriological laboratories to identify Salmonella spp.,
Shigella spp. All specimens in our study were negative for these bacterial and parasitic pathogens.
The biofilm formation was evaluated by the growth rate of E.coli on plastic surface. PCR assays
were used to detect genes of five types of diarrheagenic E.coli (DEC). All of the DEC strains showed
mannose-resistant adherence to HEp-2 cells, and aggregative adherence was predominant in these
isolates. Bacterial susceptibility to antimicrobial agents determined by the Kirby Bauer disk diffusion
method on Muller Hinton agar.
Results:
EAEC (31.9%) was the most prevalent by PCR and HEp-2 assay comparing with others. EAEC by
multiplex PCR in samples (11, 3.3%), followed by enteropathogenic E.coli (EPEC) seen in 2.1%.
Enterohemorrhagic E.coli (EHEC) and enteroinvasive E.coli (EIEC) were found in 7 (2.1%) and 1
(0.3%) of the samples. Enterotoxigenic E.coli (ETEC) and diffusely adhering E.coli were detected
in 2 (0.6%), respectively. The evaluation of bacterial biofilm formation using 96 well plates showed
309 negative (93%), 15 weak biofilm (4.6%) and 8 moderate biofilm (2.4%) formation for E.coli and
no strong biofilm forming strain was detected. Above 50% of antibiotic resistance was observed
for ampicillin, trimethoprim/sulfamethoxazole, cefuroxime and cephalotin. Also, 95.4% of isolates
were resistant to at least three different classes of antimicrobial agents and considered as multidrug
resistance.
Conclusion:
EAEC is most prevalent pathogen among DEC in our samples. It is necessary to implement EAEC
identifying method on Hep-2 assay in our laboratory practice.
2. Determination of arterial stiffness related factors in patients with type 2 diabetes
Sarantuya E ; Sainbileg S ; Uurtuya SH
Innovation 2015;9(2):62-65
Type 2 diabetes is known to be associated with elevated cardiovascular mortality and central pathological mechanism in is the process of atherosclerosis, which leads to narrowing of arterial walls throughout the body. Cardio-Ankle Vascular Index (CAVI) was developed as an index of arterial stiffness independently of early atherosclerosis. The aim of the study is to evaluate the correlation of arterial stiffness and atherosclerotic riskfactors patients with Type 2 DM. We used hospital-based onetime cross-sectional study for 52 type 2 DM patients aged 27-55 (mean age 46.7 ± 7.2) who were involved. Materials are collected by questionnaire, physicalexamination, blood analyzes and arterial stiffness is measured by VaSera VS-1000 device. Our result showed that CAVI was statistically significant on age (r=0.65 p=0.001), duration of disease (r=0.32 p=0.021), , systolic pressure (r = 0.54 p = 0.001), diastolic pressure (r = 0.54 p=0.001), red blood cell (r=0.31, p=0.02) , hematocrit (r=0.32, p=0.02) respectively. CAVI, Bodymass index and visceral fat were significantly higher in patients with hypertension than patients with normotensive.This result suggests that age, hypertension and hematocrits improve arterial stiffness in type 2diabetic patients. So it’s necessary to reduce the obesity and control the hypertension in type 2 diabetic patients to prevent from cardiovascular disease.
3.Presence and antibiotic resistance of diarrheagenic Escherichia coli in ready-to-eat salads
Unurmaa E ; Nyamtuya N ; Sarantuya J ; Tsolmon Ch ; Munkhdelger Ya
Mongolian Medical Sciences 2020;191(1):44-49
Introduction:
Foodborne diseases are a major public health concern worldwide. The report, which estimates the
burden of foodborne diseases – states that each year as many as 600 million, or almost 1 in 10
people in the world, fall ill after consuming contaminated food. Of these, 420 000 people die, including
125 000 children under the age of 5 years. The 20.3% of diarrhea and 27.5% of die caused by
contaminated foods are diarrheagenic Escherichia coli (DEC).
Aim:
To identify of DEC and determine their antibiotic resistance from ready-to-eat salads
Material and Methods:
A total of 40 bagged salad mix samples were collected from food markets in Ulaanbaatar, Mongolia.
Escherichia coli (E.coli) strains were determined on the basis of MNS 6308:2012 standard and
confirmed by polymerase chain reaction (PCR) in samples. DEC was identified using multiplex PCR.
Bacterial susceptibility to antimicrobial agents determined by the Kirby Bauer disk diffusion method.
Results:
Our results showed the presence of E. coli in 19 samples (47.5%). DEC isolates identified by multiplex
PCR were defined as follows: the presence of eae and bfp for EPEC, the presence of lt for ETEC,
the presence of ipaH for EIEC, the presence of stx1 and stx2 for EHEC, the presence of aap and
aggR for EAEC, and the presence of daaE for DAEC. The multiplex PCR assays detected EHEC 6
(31.6%), EPEC 5 (26.3%), EIEC 1 (5.3%). EAEC and ETEC were not detected in samples. The E.coli
isolates were 73.7% resistant to chloramphenicol as the first choice of treatment of diarrhea and high
resistance (68.4-94.7%) to the cephalosporins. In our country, cephalosporins are widely used in
medical practice for the treatment of infectious diseases.
Conclusion
In this study, about half of ready-to-eat salads are contaminated with E. coli. The three
types (EHEC, EPEC, EIEC) of DEC pathotypes were identified in the ready-to-eat salads and high
prevalent of antimicrobial resistance. Future research is required to track the contamination sources
and develop appropriate steps that should be taken by industry and retailers to reduce microbial
contamination in ready-to-eat salads.
4.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.
5.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.