Detection of virulence genes of Staphylococcus aureus isolated from raw beef for retail sale in the markets of Ulaanbaatar city, Mongolia.

BACKGROUND: Staphylococcus aureus (S. aureus) is a highly virulent pathogen that causes food-borne illness, food poisoning, skin and soft tissue infections, abscesses, mastitis, and bacteremia. It is common for meat and meat products to become contaminated with S. aureus due to dirty hands, food storage conditions, food production processes, and unhygienic conditions, causing food poisoning. Therefore, we aimed to isolate S. aureus strain from the raw beef and reveal virulence genes and antibiotic resistance profile from isolated S. aureus strains. METHODS: In this study, 100 samples of raw beef were collected from 4 major market stalls in Ulaanbaatar city, Mongolia. S. aureus was detected according to the ISO 6888-1:2021 standard, and the nucA gene encoding the species-specific thermonuclease was amplified and confirmed by polymerase chain reaction (PCR). In the strains of S. aureus isolated from the samples, the genes encoding the virulence factors including sea, sed, tsst, eta, etb, and mecA were amplified by multiplex PCR. These genes are encoded staphylococcal enterotoxin A, enterotoxin D, toxic shock syndrome toxin, exotoxin A, exotoxin B and penicillin-binding protein PBP 2A, respectively. Antibiotic sensitivity test was performed by the Kirby-Bauer disc diffusion method. The Clinical and Laboratory Standard Institute guidelines as CLSI M100-S27 was used for analysis of the data. RESULTS: Thirty-five percent of our samples were detected contaminated with of the S. aureus strains. Subsequently, antibiotic resistance was observed in the S. aureus contaminated samples. Among our samples, the highest rates of resistance were determined against ampicillin (97.1%), oxacillin (88.6%), and penicillin (88.6%), respectively. Three genes including mecA, sea, and tsst from six virulence genes were detected in 17% of S. aureus strain-contaminated samples by multiplex PCR. The sed, etb and eta genes were detected in the 2.9%, 11.4% and 5.7% of our samples, respectively. CONCLUSION: The results show that S. aureus related contamination is high in the raw beef for retail sale and prevalent S. aureus strains are resistant to all antibiotics used. Also, our results have demonstrated that there is a high risk for food poisoning caused by antibiotic resistant S. aureus in the raw beef and it may establish public health issues. Genes encoding for both heat-resistant and nonresistant toxicity factors were detected in the antibiotic resistant S. aureus strains and shown the highly pathogenic. Finally, our study is ensuring to need proper hygienic conditions during beef's preparation and sale.

Genetic Mutations Associated with Isoniazid Resistance in Mycobacterium tuberculosis in Mongolia.

Globally, mutations in the katG gene account for the majority of isoniazid-resistant strains of Mycobacterium tuberculosis Buyankhishig et al. analyzed a limited number of Mycobacterium tuberculosis strains in Mongolia and found that isoniazid resistance was mainly attributable to inhA mutations (B. Buyankhishig, T. Oyuntuya, B. Tserelmaa, J. Sarantuya, et al., Int J Mycobacteriol 1:40-44, 2012, https://doi.org/10.1016/j.ijmyco.2012.01.007). The GenoType MTBDRplus assay was performed for isolates collected in the First National Tuberculosis Prevalence Survey and the Third Anti-Tuberculosis Drug Resistance Survey to investigate genetic mutations associated with isoniazid resistance in Mycobacterium tuberculosis in Mongolia. Of the 409 isoniazid-resistant isolates detected by the GenoType MTBDRplus assay, 127 (31.1%) were resistant to rifampin, 294 (71.9%) had inhA mutations without katG mutations, 113 (27.6%) had katG mutations without inhA mutations, and 2 (0.5%) had mutations in both the inhA and katG genes. Of the 115 strains with any katG mutation, 114 (99.1%) had mutations in codon 315 (S315T). Of the 296 strains with any inhA mutation, 290 (98.0%) had a C15T mutation. The proportions of isoniazid-resistant strains with katG mutations were 25.3% among new cases and 36.2% among retreatment cases (P = 0.03) and 17.0% among rifampin-susceptible strains and 52.8% among rifampin-resistant strains (P < 0.01). Rifampin resistance was significantly associated with the katG mutation (adjusted odds ratio, 5.36; 95% confidence interval [CI], 3.3 to 8.67, P < 0.001). Mutations in inhA predominated in isoniazid-resistant tuberculosis in Mongolia. However, the proportion of katG mutations in isolates from previously treated cases was higher than in those from new cases, and the proportion in cases with rifampin resistance was higher than in cases without rifampin resistance.

Alteration of oral flora in Mongolian patients with Behçet's disease: a multicentre study.

OBJECTIVES: Behçet's disease (BD) is characterised by repeated acute inflammatory attacks with aphthous ulcers of the oral mucosa, uveitis of the eyes, skin symptoms, and genital ulcers. Although its aetiology is still unknown, there is evidence of the involvement of oral bacteria in systemic diseases. Various types of oral bacteria may be involved in the development and progression of BD. The present study investigated alterations in the oral flora of patients with BD in Mongolia. We collected saliva samples from the Mongolian BD group and healthy control (HC) group, and the oral flora were analysed using next-generation sequencer (NGS). METHODS: DNA was extracted from the unstimulated saliva samples from the 47 BD and 48 HC subjects. The DNA was amplified from the V3-V4 region of 16S rRNA using PCR, and the data were acquired using NGS. Based on the obtained data, we analysed the alpha diversity, beta diversity, and bacterial taxonomy of the salivary flora. RESULTS: Beta diversity differed significantly between the BD and HC flora, but no significant differences were observed in alpha diversity. We found that the proportions of three genera - an S24-7 family unknown species, a mitochondria family unknown species, and Akkermansia species associated with IL-10 production - were significantly lower in the BD than in the HC group. CONCLUSIONS: The reduced proportions of the S24-7 family and symbiotic Akkermansia species may be key phenomena in the oral flora of patients with BD.

SNP-SNP positive interaction between MMP2 and MMP12 increases the risk of COPD.

Determining SNP-SNP interaction of the disease has become important for further investigation of pathogenesis and experimental research. Although many studies have been published on the effect of MMPs gene polymorphisms on chronic obstructive pulmonary disease (COPD), there is a lack of information on SNP-SNP and SNP-environment interactions. This study aimed to investigate the interaction between the polymorphisms of MMP1, MMP2, MMP9 and MMP12 genes and its combined effect with smoking on the risk of developing COPD. Totally 181 COPD patients and 292 healthy individuals were involved. Blood samples from the participants were tested for genotyping and data were collected through questionnaires. Genotyping was performed with nested allele-specific polymerase chain reaction (AS-PCR) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). SNP-SNP and SNP-environment interactions were investigated using multifactor dimensionality reduction and logistic regression analysis. The result showed that participants with high nicotine dependence and heavy smokers had a higher risk of COPD than non-smokers. Also, G/G genotype (cOR = 5.83; 95% CI, 1.19-28.4, p = 0.029) of MMP2 rs243864 and T/T genotype (cOR = 1.79; 95% CI, 1.16-2.76, p = 0.008) of MMP12 rs652438 independently contributes to the susceptibility of COPD. For SNP-SNP interaction, the positive interaction between rs243864 G/G genotype of MMP2 and rs652438 T/T genotype of MMP12 was found, and the combination of risk genotypes has a high risk of COPD (OR = 12.92; 95% CI, 1.46-114.4, p = 0.021). Moreover, the combination of T/T genotype of MMP12 rs652438 and smoking-related factors increases the risk of COPD approximately 4.5 to 6-fold. The results suggests that there is a combination of MMP2, MMP12, and smoking-related factors may increase the risk of developing COPD.

Detection and antibiotic resistance of diarrheagenic Escherichia coli from patients with diarrhea in Ulaanbaatar, Mongolia.

INTRODUCTION: Diarrheal diseases are common with worldwide distribution, and diarrheagenic Escherichia coli (DEC) strains are the main causative agents. The present study aimed to define the association of various pathotypes of E. coli from diarrheal patients in Mongolia. METHODOLOGY: A total of 341 E. coli strains were isolated from the stool of diarrheal patients. Bacterial susceptibility to antimicrobial agents was determined by the Kirby Bauer disk diffusion method. DEC isolates were identified by HEp-2 cell adherence assay and multiplex polymerase chain reaction (PCR). RESULTS: DEC pathogens were detected in 53.7% of 341 E. coli isolates. Enteroaggregative E. coli (EAEC) was the most common DEC pathotype identified by HEp-2 adherence assay and multiplex PCR methods in 97 samples (28.4%), followed by atypical enteropathogenic E. coli (EPEC) in 50 samples (14.7%), diffusely adherent E. coli (DAEC) in 25 samples (7.3%), enterohaemorrhagic E. coli (EHEC) in 6 samples (1.8%), enterotoxigenic E. coli (ETEC) in 4 samples (1.2%), and enteroinvasive E. coli (EIEC) in 1 sample (0.3%). DEC strains had > 50% antibiotic resistance against cephalothin, ampicillin, and trimethoprim/sulfamethoxazole. All tested DEC strains were susceptible to imipenem. Among the 183 DEC strains, 27 (14.8%) were extended spectrum beta-lactamase producing isolates, and 125 (68.3%) isolates were multiple drug resistant. CONCLUSIONS: We have identified six pathotypes of DEC from the clinical isolates tested and concluded that a high prevalence of antimicrobial resistance was observed in these pathotypes. EAEC was the most common pathotype identified and this is the first report of EHEC identification in Mongolia.

Association of clinical course with thyroid-stimulating immunoglobulin in Graves' ophthalmopathy in Mongolians.

Graves' ophthalmopathy (GO) is a complex inflammatory condition affecting the orbit and is often associated with Graves' disease (GD). This study aims to determine the levels of thyroid-stimulating immunoglobulin (TSI) and thyroid-stimulating hormone receptor autoantibody (TSHR-ab) in the serum of patients with GO, compare it with the GD, and determine whether there is a correlation with the clinical course of GO. The cross-sectional study included 82 patients with GO, 81 patients with GD, and 75 healthy subjects. The ocular manifestations of GO were identified and evaluated by the clinical activity score (CAS) and severity of GO using the European Group of Graves' Orbitopathy (EUGOGO). TSI and TSHR-ab levels in the serum of participants were determined with ELISA kits and correlated with clinical findings. A total of 238 participant's data were analyzed. There were 14 patients (17%) with unilateral GO. The most common ocular signs were eyelid retraction 68 (82.3%) and proptosis 61 (74.4%). The mean CAS score was 2.65±1.64 in GO patients and was higher in men than women (P = 0.008). The mean of TSI was 37.95±35.41 in GO, 14.16±15.67 in GD, and 4.33±2.94 in healthy controls (P<0.0001). The TSI was significantly higher in patients with GO than in those with GD (P<0.0001). There were no correlations between TSI and TSHR-ab levels and CAS scores. However, we observed a correlation between the TSI level and the severity of GO (P = 0.023). The area under the ROC curve (AUC) of TSI was 0.933 and selected 14.1 IU/ml was the optimal cutoff value (98.78% of sensitivity, 83.97% of specificity). Our study showed that TSI is significantly related to GO and the severity of GO. Therefore, TSI can be used as a predictor of severe GO to help in prognostication, follow-up and treatment planning.

Main characteristics of dermatoglypics associated with schizophrenia and its clinical subtypes.

Dermatoglypic patterns are extensively investigated to apply in disease-related risk assessment due to an obvious association between morphological and genetic characteristics. In the current study, we aimed to determine whether the fingerprint and palmar patterns vary between case population with schizophrenia and general population. A cross sectional study was conducted in people diagnosed with schizophrenia (cases) and a control population between 2016 and 2019. In this study, 252 people were participated. Ink and paper method was used to evaluate the difference of fingerprints palm prints between patients with schizophrenia and participants in control group.93 participants were analyzed in schizophrenic group and 142 participants were investigated in the control group. The percentage of arches on the right ring finger was significantly different between the schizophrenic patient group and control group (p = 0.011). The whorl pattern type (U-W-U-W-W-W-W-U-W-U) was dominantly observed in both of the schizophrenic patient group and control group. A-B ridge count in schizophrenic patient group and control group produced a markedly significant difference (p<0.05). Interestingly, a strong significant difference was produced in comparing of A-B ridge count in catatonic schizophrenia group with residual schizophrenia group (p<0.005). In comparison, index of pattern intensity in control group was slightly higher than that in schizophrenic patient group. Taking together, these results showed that the dermatoglypic characteristics might be a valuable tool to describe the nature of schizophrenia and its clinical subtypes and further studies are needed in clinical application.

The Cumulative Effect of Gene-Gene Interactions Between GSTM1, CHRNA3, CHRNA5 and SOD3 Gene Polymorphisms Combined with Smoking on COPD Risk.

BACKGROUND: Chronic obstructive pulmonary disease (COPD) is a multifactorial disorder which is affected by external and internal risk factors. People with no external risk factors may be significantly affected and develop pulmonary disease. The study aimed to define gene-gene and gene-environmental effects on COPD. METHODS: A case control study involved 181 COPD patients and 292 healthy individuals, with peripheral blood sampling and adequate questionnaires. Genotyping was done with various types of PCR design for GSTM1 (null del), GSTT1 (null del), EPHX1 (rs2234922 and rs1051740), GSTP1 (rs1695 and rs1138272), CHRNA3 (rs1051730 and rs12914385), CHRNA5 (rs16969968 and rs17486278), and SOD3 (rs1799895 and rs699473) gene polymorphisms. Gene-gene and gene-environmental interactions were investigated using multidimensional regression analysis. RESULTS: Frequency of risk alleles of rs1051730 (p = 0.001), rs16969968 (p <0.001), and rs1799895 (p <0.001) polymorphisms were significant in univariate analysis. For gene-gene interaction, GSTM1 null, rs1051730, rs16969968, and rs1799895 polymorphisms independently contributed to risk of COPD and any combinations of the risk genotypes have a higher risk of disease. A cumulative effect of the four risk polymorphisms increased the risk of COPD for the smoking index (cOR = 13.6, p <0.001), cigarettes per day (cOR = 32.08, p <0.01), nicotine dependence (cOR = 12.0, p <0.01), and smoking status (cOR = 17.02, p <0.01) for gene-environmental interaction. CONCLUSION: Several pivotal genes showed distinct effects for COPD, and some synergistic effects affected the disease progression. The development of COPD was synergistically increased with gene-gene and gene-environmental risk factors.

Clinical features of Behcet's disease in Mongolia: a multicenter study.

OBJECTIVE: The aim of the present study is to investigate the clinical features of patients with Behcet's disease (BD) in Mongolia. METHODS: Patients were identified and examined from six medical institutions in Mongolia from January 2015 to January 2019. BD was diagnosed according to the diagnostic criteria for BD established by the International Study Group. RESULTS: There were sixty-five patients (22 males and 43 females) recoded, the ratio of 1:1.95, with a marked female predominance. The age of disease onset was 22.2 ± 10.0 (mean ± SD), ranging from 11 to 66 years old. Oral aphthous ulcers, ocular lesions, skin lesions, genital ulcers, pathergy test positivity, articular lesions, superficial vasculitis, deep vein thrombosis, and epididymitis (male only) were observed in 100.0%, 63.1%, 81.5%, 89.2%, 7.7%, 86.2%, 32.3%, 4.6%, and 13.6% of the patients, respectively. The incidence of poor visual prognosis, ≤ 20/200, was significantly higher in males than in females (31.8 vs. 9.3%, incidence rate ratio 4.55 (95% CI 1.16-17.82), p < 0.05). The pathergy test was positive only in 7.7% of cases and only in female subjects. Nasal mucous ulcers were frequently seen in 55.4% of patients that may also be attributed to the environmental conditions of Mongolia. Headache was observed 76.9% of patients in this study. CONCLUSIONS: Clinical manifestations of BD in Mongolia are presented for the first time. The visual prognosis was significantly worse in males. Nasal mucous membrane ulcers and recurrent headaches were frequent among Mongolian patients with BD. Key Points • First results of the examination of the clinical features of Behcet's disease patients in Mongolia. • Nasal ulcerations and recurrent headaches are frequent symptoms in Mongolia Behcet's disease patients, potentially attributed to climate. • Male Behcet's disease patients in Mongolia have a significantly worse prognosis for eye-related complications and vision.

Influenza B viruses circulated during last 5 years in Mongolia.

Influenza B virus-caused illness has recently been considered as an urgent public health problem due to substantial morbidity, mortality and life-threatening medical complications. In this study, we have reported the main characteristics of influenza B virus in Mongolia, including prevalence, lineages, suitability with vaccine strains and drug susceptibility against the virus. 15768 specimens were tested by qPCR for detecting influenza viruses. From positive specimens for influenza B virus, the clinical isolates were isolated using MDCK cells. Sequencing analysis, hemagglutination inhibition assay and Neuraminidase inhibitor (NAI) drug susceptibility testing were performed for the clinical isolates. Influenza B virus was around in 3.46% of the samples in Mongolia, and B/Victoria clade-1A and B/Yamagata clade-3 lineages were predominant. Importantly, it was confirmed that the lineages corresponded to the vaccine strains. Moreover, drug susceptibility tests revealed that some Mongolian clinical isolates showed reduced susceptibility to antiviral agents. Interestingly, G104R was identified as a novel mutation, which might have a significant role in drug resistance of the virus. These results describe the characteristics of influenza B viruses that have caused respiratory illness in the population of Mongolia between 2013 and 2017.

Antibiotic resistance of Helicobacter pylori in Mongolia.

INTRODUCTION: The resistance of Helicobacter pylori to recently available antibiotic treatment regimens has been recognized as a growing problem. Therefore, the aim of this study was to determine the prevalence of antibiotic resistance among H. pylori strains isolated from Mongolians. METHODOLOGY: All gastric biopsy specimens were obtained during upper gastrointestinal endoscopy from patients referred for the exploration of dyspepsia. The urease positive samples by rapid urease test were cultured according to standard microbiological procedures and H. pylori were grown under microaerophilic conditions on selective Pylori agar. H. pylori antibiotic sensitivity was examined using E-test. In addition, the mutations of the corresponding gene were studied by GenoType HelicoDR DNA strip testing. RESULTS: Three hundred twenty patients, 216 female and 104 male in the ages range of 18 to 83 years were included in this study. Rapid urease test yielded positive results for 65.9% (211/320). Among them, we have successfully obtained 72% H. pylori isolates. The antibiotic resistance rates were 35.5% for clarithromycin, 68.4% metronidazole, 23.0% amoxicillin, 25.0% tetracycline, 28.2% erythromycin and 14.5% nitrofuranton. Resistance for 2 drugs was 34.5% and that of 3 drugs was observed in 14.5% of isolates. The most prevalent mutation was A2147G followed by A2146G and D91Y. The prevalence of H. pylori infection increased among Mongolian population and the prevalence of resistance of H. pylori is very high to metronidazole, and moderate to clarithromycin. CONCLUSION: The data on antimicrobial susceptibilities provided by the present study is may assist the clinicians on the effectiveness of treatment regimens.

Correlation between Reproductive Hormonal Level and Osteoporosis among Women in Mongolia.

BACKGROUND: Postmenopausal osteoporosis is the most common bone metabolic disease associated with low bone mineral density (BMD) and osteopathic fragility fractures, which can lead to significant morbidity. The objective of this study was to investigate the relationship between serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E2) levels and bone mineral density (BMD) across the stages of menopause in Mongolian women. METHODS: Two hundred sixty participants aged 50.1±4.4 years were enrolled in the study. Blood samples were obtained from each participant and analyzed using ELISA. Data were first stratified and analyzed by bone mineral density status (osteoporotic, osteopenic, and normal) and then by menopause status. Between group differences were analyzed using t-tests, and correlations were assessed using the Spearman rank order test, with Bonferonni correction. The data were analyzed using Statistical Package Statistical Software version 20.0 (SPSS Inc., Chicago, IL). Significance was set at p<0.05. RESULTS: The mean menopausal age was 48.4±3.4, which is comparable to the Mongolian population mean menopausal age. The mean serum estradiol level in the normal BMD group was 18.3±13.1 pg/ml and 15.8±10.7 pg/ml in the osteoporotic group. The mean serum FSH in the normal BMD group was 54.5±44.1 pg/ml and 81.3±34.2 pg/ml in the osteoporotic group. The mean serum LH level in the normal BMD group was 53.1±41.2 and 75.1±26.1 pg/ml in the osteoporotic group. The mean T and Z score were lower in the osteoporotic group. FSH and LH levels significantly differed across menopause stages in that those who were post-menopausal had higher levels compared to those who were pre- or peri-menopausal. Both hormones, FSH and LH, showed weak negative correlations with BMD level, but not E2. There were significant negative correlations between FSH and Speed of Sound (SOS) (r=-0.16; p<0.01), and between osteoporosis with age (r=-0.30, p<0.05) and number of childbirths (r=-0.14 p<0.05). DISCUSSION: Osteoporosis is a significant problem with associations to hormone levels in post-menopausal women. In our study, mean serum estradiol levels decreased with age, and the mean FSH and LH levels were higher in women of later menopausal stage. Further study is warranted to investigate the bone related studies to establish better statistical references among Mongolian women.