Gülnihan Üstündağ, Aslıhan Şahin, Yücehan Yılmaz Yazıcı, Ahu Kara Aksay, Can Biçmen, Serkan Belkaya, Dilek Yılmaz
Bovine tuberculosis might be seen in low-income countries, especially in children fed with raw milk. The most common transmission route is fecal-oral way, and it is most likely through unpasteurized dairy products. Although clinical and radiological findings are like non-zoonotic tuberculosis, treatment approaches may differ in individuals with zoonotic tuberculosis. Prevention of zoonotic diseases requires multidisciplinary approaches. These approaches include the development of veterinary and surveillance studies for the detection of communicable diseases in farm animals, as well as informing the public about raw milk consumption. In this case report, a patient with zoonotic pulmonary tuberculosis related to Mycobacterium bovis because of consumption of raw milk was presented. A five-month-old male was admitted to the hospital due to a persistent, feverless, non-productive cough since birth. Empirical antibiotic treatment was started with a preliminary diagnosis of pneumonia because of left upper lobe and right pericardial infiltration on chest X-ray. However, after two weeks of antimicrobial therapy, the patient's clinical and laboratory findings did not improve. This led to the referral for a computed tomography imaging, which revealed tracheomalacia, consolidation on the right upper lobe, an indistinguishable mass or consolidation on the left middle lobe of the lung, peribronchial thickening on the basal segment of the lower lobe, and mediastinal lymphadenopathy. Three consecutive days of fasting gastric lavage fluid was sent to the reference laboratory for acid-resistant bacillus examination, polymerase chain reaction (PCR) and culture studies. As the clinical findings were compatible and PCR was positive, the patient was started on quadruple antituberculous therapy. After initiation of anti-tuberculosis drugs, the patient's findings radiologically and clinically were improved. Mycobacterium bovis was grown in the culture. In the meantime, it was discovered that the patient was fed with raw milk. Due to the patient's clinical symptoms and the growth of Mycobacterium bovis in the gastric lavage fluid culture, the diagnosis of bovine tuberculosis was made. The culprit was that the milk of the cow belonging to the patient's family, which was later found to be infected with M.bovis, was milked and given to the patient without boiling. Today, unpasteurized dairy products continue to be consumed, especially in rural areas. One of the most important steps to prevent zoonotic diseases is to raise awareness about not consuming raw milk and undercooked meat. To elucidate the epidemiological link in childhood, taking a good anamnesis, including questioning raw milk consumption, is essential in the diagnosis of tuberculosis.
{"title":"[An Infant with Zoonotic Pulmonary Tuberculosis due to Mycobacterium bovis].","authors":"Gülnihan Üstündağ, Aslıhan Şahin, Yücehan Yılmaz Yazıcı, Ahu Kara Aksay, Can Biçmen, Serkan Belkaya, Dilek Yılmaz","doi":"10.5578/mb.20239939","DOIUrl":"https://doi.org/10.5578/mb.20239939","url":null,"abstract":"<p><p>Bovine tuberculosis might be seen in low-income countries, especially in children fed with raw milk. The most common transmission route is fecal-oral way, and it is most likely through unpasteurized dairy products. Although clinical and radiological findings are like non-zoonotic tuberculosis, treatment approaches may differ in individuals with zoonotic tuberculosis. Prevention of zoonotic diseases requires multidisciplinary approaches. These approaches include the development of veterinary and surveillance studies for the detection of communicable diseases in farm animals, as well as informing the public about raw milk consumption. In this case report, a patient with zoonotic pulmonary tuberculosis related to Mycobacterium bovis because of consumption of raw milk was presented. A five-month-old male was admitted to the hospital due to a persistent, feverless, non-productive cough since birth. Empirical antibiotic treatment was started with a preliminary diagnosis of pneumonia because of left upper lobe and right pericardial infiltration on chest X-ray. However, after two weeks of antimicrobial therapy, the patient's clinical and laboratory findings did not improve. This led to the referral for a computed tomography imaging, which revealed tracheomalacia, consolidation on the right upper lobe, an indistinguishable mass or consolidation on the left middle lobe of the lung, peribronchial thickening on the basal segment of the lower lobe, and mediastinal lymphadenopathy. Three consecutive days of fasting gastric lavage fluid was sent to the reference laboratory for acid-resistant bacillus examination, polymerase chain reaction (PCR) and culture studies. As the clinical findings were compatible and PCR was positive, the patient was started on quadruple antituberculous therapy. After initiation of anti-tuberculosis drugs, the patient's findings radiologically and clinically were improved. Mycobacterium bovis was grown in the culture. In the meantime, it was discovered that the patient was fed with raw milk. Due to the patient's clinical symptoms and the growth of Mycobacterium bovis in the gastric lavage fluid culture, the diagnosis of bovine tuberculosis was made. The culprit was that the milk of the cow belonging to the patient's family, which was later found to be infected with M.bovis, was milked and given to the patient without boiling. Today, unpasteurized dairy products continue to be consumed, especially in rural areas. One of the most important steps to prevent zoonotic diseases is to raise awareness about not consuming raw milk and undercooked meat. To elucidate the epidemiological link in childhood, taking a good anamnesis, including questioning raw milk consumption, is essential in the diagnosis of tuberculosis.</p>","PeriodicalId":18509,"journal":{"name":"Mikrobiyoloji bulteni","volume":"57 3","pages":"473-480"},"PeriodicalIF":1.0,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9831695","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Yeliz Tanrıverdi Çaycı, Kübra Hacıeminoğlu Ülker, Gülşah Karacan Temür, Doğa Beliz Güney, Mustafa Ertokatlı, Asuman Birinci
Escherichia coli ST131 clone was first reported in 2008 and defined as a 'high-risk pandemic clone'. This clone plays a critical role in the spread of antimicrobial resistance worldwide. It was reported in many studies that the E.coli ST131 clone is widespread worldwide and can carry virulence and antibiotic resistance genes. E.coli ST131 clone is associated with a fluoroquinolone and broad-spectrum cephalosporin resistance. The agents used in the treatment of infections caused by the E.coli ST131 clone are limited. For this reason, monitoring the resistance status of these limited agents is crucial. In this study, we aimed to investigate the prevalence of the O25b-ST131 clone, and the presence of carbapenem and fosfomycin resistance genes in fluoroquinolone-resistant E.coli isolates isolated from mid-stream urine samples. For the detection of the O25b-ST131 clone in fluoroquinolone-resistant E.coli isolates, amplification was performed with primers O25pabBspe-F and O25pabBspe-R. For the determination of resistance genes, carbapenem resistance genes blaNDM, blaVIM, blaKPC, blaIMP, and blaOXA-48 in carbapenem resistant isolates and plasmid-mediated fosfomycin resistance genes fosA3 and fosC2 in fosfomycin resistant isolates were investigated by multiplex PCR method. According to PCR results, the prevalence of E.coli O25b-ST131 isolates was 51.2%. Carbapenem resistance rate was 3.41%, and fosfomycin resistance rate was 3.41% in fluoroquinolone-resistant E.coli isolates. Carbapenem and fosfomycin resistance rates in E.coli O25bST131 isolates were determined as 0.83% and 2.5%, respectively. At least one of the carbapenem-resistance genes (blaNDM and/or blaOXA-48) was detected in six of the eight carbapenem-resistant isolates. Fosfomycin resistance gene fosA3 was seen in four of eight fosfomycin-resistant isolates. fosC2 gene was not detected in any of the isolates. In addition, the plasmid-mediated fosfomycin resistance gene fosA3 was detected in an E.coli O25b-ST131 isolate, and this result was confirmed by sequence analysis. To the best of our knowledge, this is the second report about fosA3 positivity in E.coli ST131 isolates from the world and the first reported from Europe and Türkiye. As a result, approximately half of the fluoroquinolone-resistant E.coli isolates were identified as E.coli ST131 clones. Despite the high rate of this clone, the carbapenem and fosfomycin resistance rates are still relatively low, which is pleasing for the future of treatment. However, it should not be forgotten that resistance rates and the prevalence of resistance genes should be constantly monitored.
{"title":"[Investigation of O25b-ST131 Clone Frequency and Presence of Carbapenem and Fosfomycin Resistance Genes in Escherichia coli Isolates: First Detection of fosA3 from Escherichia coli O25bSTf131 Clone from Türkiye].","authors":"Yeliz Tanrıverdi Çaycı, Kübra Hacıeminoğlu Ülker, Gülşah Karacan Temür, Doğa Beliz Güney, Mustafa Ertokatlı, Asuman Birinci","doi":"10.5578/mb.20239937","DOIUrl":"https://doi.org/10.5578/mb.20239937","url":null,"abstract":"<p><p>Escherichia coli ST131 clone was first reported in 2008 and defined as a 'high-risk pandemic clone'. This clone plays a critical role in the spread of antimicrobial resistance worldwide. It was reported in many studies that the E.coli ST131 clone is widespread worldwide and can carry virulence and antibiotic resistance genes. E.coli ST131 clone is associated with a fluoroquinolone and broad-spectrum cephalosporin resistance. The agents used in the treatment of infections caused by the E.coli ST131 clone are limited. For this reason, monitoring the resistance status of these limited agents is crucial. In this study, we aimed to investigate the prevalence of the O25b-ST131 clone, and the presence of carbapenem and fosfomycin resistance genes in fluoroquinolone-resistant E.coli isolates isolated from mid-stream urine samples. For the detection of the O25b-ST131 clone in fluoroquinolone-resistant E.coli isolates, amplification was performed with primers O25pabBspe-F and O25pabBspe-R. For the determination of resistance genes, carbapenem resistance genes blaNDM, blaVIM, blaKPC, blaIMP, and blaOXA-48 in carbapenem resistant isolates and plasmid-mediated fosfomycin resistance genes fosA3 and fosC2 in fosfomycin resistant isolates were investigated by multiplex PCR method. According to PCR results, the prevalence of E.coli O25b-ST131 isolates was 51.2%. Carbapenem resistance rate was 3.41%, and fosfomycin resistance rate was 3.41% in fluoroquinolone-resistant E.coli isolates. Carbapenem and fosfomycin resistance rates in E.coli O25bST131 isolates were determined as 0.83% and 2.5%, respectively. At least one of the carbapenem-resistance genes (blaNDM and/or blaOXA-48) was detected in six of the eight carbapenem-resistant isolates. Fosfomycin resistance gene fosA3 was seen in four of eight fosfomycin-resistant isolates. fosC2 gene was not detected in any of the isolates. In addition, the plasmid-mediated fosfomycin resistance gene fosA3 was detected in an E.coli O25b-ST131 isolate, and this result was confirmed by sequence analysis. To the best of our knowledge, this is the second report about fosA3 positivity in E.coli ST131 isolates from the world and the first reported from Europe and Türkiye. As a result, approximately half of the fluoroquinolone-resistant E.coli isolates were identified as E.coli ST131 clones. Despite the high rate of this clone, the carbapenem and fosfomycin resistance rates are still relatively low, which is pleasing for the future of treatment. However, it should not be forgotten that resistance rates and the prevalence of resistance genes should be constantly monitored.</p>","PeriodicalId":18509,"journal":{"name":"Mikrobiyoloji bulteni","volume":"57 3","pages":"454-462"},"PeriodicalIF":1.0,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10239541","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Cankut Karabulut, Tülay Aksoy, Ahmet Yıldırım, I Cüneyt Balcıoğlu
Leishmania RNA virus (LRV) is a double-stranded RNA (dsRNA) virus that is thought to contribute to the severe inflammatory response of the causative Leishmania parasite in the mammalian host by being present in many isolates of Leishmania spp. In our study, it was aimed to obtain data on the presence of Leishmania RNA Virus 2 (LRV2), which is thought to cause a change in the clinical course of leishmaniasis, in Leishmania major and Leishmania tropica isolates isolated from cutaneous leishmaniasis (CL) patients in Türkiye. Leishmania strains stored in liquid nitrogen tank by cryopreservation in Manisa Celal Bayar University Faculty of Medicine Parasite Bank were resuscitated under suitable conditions and cultivated in NNN and RPMI-1640 media. Then, the isolates were allowed to enter the logarithmic phase in a 26ºC incubator and DNA isolations were made using the "High Pure PCR Template Preparation Kit". Real-time polymerase chain reaction (Rt-PCR) melting analyzes were applied to the DNAs obtained by using primers and probes specific to the internal transcribed spacer-1 (ITS-1) gene region of Leishmania. After RNA isolation from promastigote suspension, cDNA synthesis was performed by reverse transcription. After gel electrophoresis with PCR amplification products, dsRNA band formation was evaluated in terms of LRV2 positivity under ultraviolet light. Among the 20 examined Leishmania spp. isolates (10 L.tropica and 10 L.major), four (three L.tropica, one L.major) were found to be positive for the presence of LRV2. Although the mechanism of LRV in recent studies has not been fully understood, it is known that it exacerbates the clinic of the disease and even has an effect on the formation of drug resistance by the parasite. It is important to obtain data on the presence of LRV in our country and to contribute to various clinical, drug development, prevalence studies, diagnosis and treatment of the disease in the future.
{"title":"[Investigation of Leishmania RNA Virus 2 in Leishmania major and Leishmania tropica Strains Isolated from Cutaneous Leishmaniasis Patients in Türkiye].","authors":"Cankut Karabulut, Tülay Aksoy, Ahmet Yıldırım, I Cüneyt Balcıoğlu","doi":"10.5578/mb.20239938","DOIUrl":"https://doi.org/10.5578/mb.20239938","url":null,"abstract":"<p><p>Leishmania RNA virus (LRV) is a double-stranded RNA (dsRNA) virus that is thought to contribute to the severe inflammatory response of the causative Leishmania parasite in the mammalian host by being present in many isolates of Leishmania spp. In our study, it was aimed to obtain data on the presence of Leishmania RNA Virus 2 (LRV2), which is thought to cause a change in the clinical course of leishmaniasis, in Leishmania major and Leishmania tropica isolates isolated from cutaneous leishmaniasis (CL) patients in Türkiye. Leishmania strains stored in liquid nitrogen tank by cryopreservation in Manisa Celal Bayar University Faculty of Medicine Parasite Bank were resuscitated under suitable conditions and cultivated in NNN and RPMI-1640 media. Then, the isolates were allowed to enter the logarithmic phase in a 26ºC incubator and DNA isolations were made using the \"High Pure PCR Template Preparation Kit\". Real-time polymerase chain reaction (Rt-PCR) melting analyzes were applied to the DNAs obtained by using primers and probes specific to the internal transcribed spacer-1 (ITS-1) gene region of Leishmania. After RNA isolation from promastigote suspension, cDNA synthesis was performed by reverse transcription. After gel electrophoresis with PCR amplification products, dsRNA band formation was evaluated in terms of LRV2 positivity under ultraviolet light. Among the 20 examined Leishmania spp. isolates (10 L.tropica and 10 L.major), four (three L.tropica, one L.major) were found to be positive for the presence of LRV2. Although the mechanism of LRV in recent studies has not been fully understood, it is known that it exacerbates the clinic of the disease and even has an effect on the formation of drug resistance by the parasite. It is important to obtain data on the presence of LRV in our country and to contribute to various clinical, drug development, prevalence studies, diagnosis and treatment of the disease in the future.</p>","PeriodicalId":18509,"journal":{"name":"Mikrobiyoloji bulteni","volume":"57 3","pages":"463-472"},"PeriodicalIF":1.0,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9831698","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Ebru Yücebağ, Neşe Arslan, Yeşim Tok, Okan Kadir Nohut, Seda Salman Yılmaz, Mert Ahmet Kuşkucu, Kenan Midilli
<p><p>Coronavirus disease-2019 (COVID-19) emerged in the last months of 2019 and caused a pandemic effecting the whole world. Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), the causative agent of COVID-19 has changed by various mutations since the day it was first identified, causing the pandemic to continue. Age, male gender, obesity, and comorbidity, which are general risk factors for COVID-19, can also cause prolonged PCR positivity. In this report, a case of 37-year-old male who is working in the hospital's COVID-19 molecular diagnostics laboratory was presented. He was vaccinated with three doses of inactivated vaccine, CoronaVac (Sinovac Biotech, Beijing-China), within the context of the vaccination program carried out in Türkiye. His first SARS-CoV-2 positivity was detected on 12.01.2021, four months after the last vaccination, and he continued to be detected positive for SARS-CoV-2 throughout a period of 39 days by quantitative reverse transcription polymerase chain reaction (qRT-PCR) tests performed with 2-3-day intervals. The patient has a 20-pack/year smoking history and his body mass index (BMI) was 29.8 kg/m2 at the time of his COVID-19. The case, which was clinically defined as mild COVID-19 with symptoms including back and headache, cough, fever (38.5°C), and loss of taste-smell, and without any additional complications or respiratory distress during the disease process. In the radiological examination, the lung was found within normal ranges. Prophylactic enoxaparin sodium anti-xa IU/0.6 ml was administered to the patient due to his cardiovascular risk, and no additional treatment was given. Whole genome sequencing was performed from nasopharyngeal swab samples of the patient at the beginning and 16th day of the infection to investigate the the specific genomic features and mutation pattern of the virus in the host over time, due to the prolonged SARS-CoV-2 PCR positivity. Library preparation for the whole next-generation sequencing (NGS) was performed by the SARS-CoV-2 Panel, Paragon CleanPlex kit (Paragon Genomics, USA), and indexing of the library was done by Clean-Plex Dual-Indexed PCR Primers for Illumina Set B kit (Paragon Genomics, USA). NGS analysis was performed on the Illumina Miniseq (Illumina, USA) platform. As a result of the bioinformatics evaluation, both samples were determined as SARS-CoV-2 Delta variant (Nextclade; 21J-Delta variant, Pango lineage; AY.43). Remarkably, the SARSCoV-2 sequences in the two samples taken 15 days apart; several identical mutations; such as D614G in the S gene, P323L in the ORF 1b gene region, and P1228L in the Nsp3 gene region, were detected. Besides that, when compared to the first sample, three additional mutations (P383L, P539S, L838I) were observed in the sequence of the second sample, which led to three amino acid changes, the clinical significance of which has not yet been determined in the literature. It is thought that; these mutations that change amino acid expr
{"title":"[P323L Mutation in a Case with Prolonged SARS-CoV-2 PCR Positivity].","authors":"Ebru Yücebağ, Neşe Arslan, Yeşim Tok, Okan Kadir Nohut, Seda Salman Yılmaz, Mert Ahmet Kuşkucu, Kenan Midilli","doi":"10.5578/mb.20239941","DOIUrl":"https://doi.org/10.5578/mb.20239941","url":null,"abstract":"<p><p>Coronavirus disease-2019 (COVID-19) emerged in the last months of 2019 and caused a pandemic effecting the whole world. Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), the causative agent of COVID-19 has changed by various mutations since the day it was first identified, causing the pandemic to continue. Age, male gender, obesity, and comorbidity, which are general risk factors for COVID-19, can also cause prolonged PCR positivity. In this report, a case of 37-year-old male who is working in the hospital's COVID-19 molecular diagnostics laboratory was presented. He was vaccinated with three doses of inactivated vaccine, CoronaVac (Sinovac Biotech, Beijing-China), within the context of the vaccination program carried out in Türkiye. His first SARS-CoV-2 positivity was detected on 12.01.2021, four months after the last vaccination, and he continued to be detected positive for SARS-CoV-2 throughout a period of 39 days by quantitative reverse transcription polymerase chain reaction (qRT-PCR) tests performed with 2-3-day intervals. The patient has a 20-pack/year smoking history and his body mass index (BMI) was 29.8 kg/m2 at the time of his COVID-19. The case, which was clinically defined as mild COVID-19 with symptoms including back and headache, cough, fever (38.5°C), and loss of taste-smell, and without any additional complications or respiratory distress during the disease process. In the radiological examination, the lung was found within normal ranges. Prophylactic enoxaparin sodium anti-xa IU/0.6 ml was administered to the patient due to his cardiovascular risk, and no additional treatment was given. Whole genome sequencing was performed from nasopharyngeal swab samples of the patient at the beginning and 16th day of the infection to investigate the the specific genomic features and mutation pattern of the virus in the host over time, due to the prolonged SARS-CoV-2 PCR positivity. Library preparation for the whole next-generation sequencing (NGS) was performed by the SARS-CoV-2 Panel, Paragon CleanPlex kit (Paragon Genomics, USA), and indexing of the library was done by Clean-Plex Dual-Indexed PCR Primers for Illumina Set B kit (Paragon Genomics, USA). NGS analysis was performed on the Illumina Miniseq (Illumina, USA) platform. As a result of the bioinformatics evaluation, both samples were determined as SARS-CoV-2 Delta variant (Nextclade; 21J-Delta variant, Pango lineage; AY.43). Remarkably, the SARSCoV-2 sequences in the two samples taken 15 days apart; several identical mutations; such as D614G in the S gene, P323L in the ORF 1b gene region, and P1228L in the Nsp3 gene region, were detected. Besides that, when compared to the first sample, three additional mutations (P383L, P539S, L838I) were observed in the sequence of the second sample, which led to three amino acid changes, the clinical significance of which has not yet been determined in the literature. It is thought that; these mutations that change amino acid expr","PeriodicalId":18509,"journal":{"name":"Mikrobiyoloji bulteni","volume":"57 3","pages":"490-497"},"PeriodicalIF":1.0,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9834220","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
<p><p>The rate of extensively drug-resistant and pan-resistant gram-negative rods isolated as infectious agents is increasing around the world and in Türkiye. One of the important options in the treatment of these infections is the combined use of antibiotics. Therefore, the aim of this study was to investigate the in vitro effect of meropenem/colistin and meropenem/fosfomycin combinations on carbapenem-resistant gram-negative bacilli isolated as infectious agents. Escherichia coli (n= 6), Klebsiella pneumoniae (n= 10), Pseudomonas aeruginosa (n= 5), and Acinetobacter baumannii (n= 6) isolates were recovered from blood and tracheal aspirate samples of patients hospitalized in our hospital's intensive care unit were included in the study. In the first stage of the combination study, minimal inhibitory concentrations (MIC) were investigated by broth microdilution for meropenem and colistin, and agar dilution methods for fosfomycin. In the second stage of the study, synergy, partial synergy, indifference, and antagonistic effects were investigated with the checkerboard method for the meropenem/colistin combination and the agar dilution method for the meropenem/fosfomycin combination. The checkerboard results were interpreted as follows: fractional inhibitory concentration index (FICI) values ≤ 0.5 synergy, < 0.5-≤ 1 partial synergy, > 1-≤ 4 indifference and FIC values of > 4 antagonism. MIC values obtained in the study were interpreted according to European Committee on Antimicrobial Susceptibility Testing (EUCAST) criteria. Of the 27 isolates studied with the broth microdilution method, 63% were found to be colistin-resistant and 37% susceptible. The MIC values of fosfomycin against Enterobacterales group bacteria were found to be in the range of 2-2048 mg/L. Two of the six E.coli isolates and nine of the 10 K.pneumoniae isolates were found to be resistant to fosfomycin (IV). The MIC values of ≥ 128 mg/L were found in all 11 non-fermentative gram-negative rods with intrinsic resistance to fosfomycin. In the combination of meropenem/ colistin, synergy and partial synergy were observed in 11 (40.7%) of 27 isolates, an indifference effect was observed in 13 (48.2%), and antagonistic effects were observed in three (11.1%) of the isolates. The synergy and partial synergy effects of this combination were 37.5% for Enterobacterales group bacteria, 50% for E.coli, and 30% for K.pneumoniae. Regarding the 11 non-fermentative gram-negative rods included in the study, 83.3% synergy and partial synergy was found in A.baumannii for the meropenem/colistin combination, while no synergy and partial synergistic effect was found in P.aeruginosa. Meropenem/fosfomycin synergy and partial synergy effects were 83.3% (5/6) for E.coli, 100% (8/8) for K.pneumoniae, 100% (6/6) for A.baumannii, and 25% (1/4) for P.aeruginosa. In all of the isolates studied, meropenem/fosfomycin combination was found to be more effective than the meropenem/colistin combination. It would be mea
{"title":"[Evaluation of In vitro Efficacy of Meropenem/Colistin and Meropenem/Fosfomycin Combinations on Multidrug Resistant Gram-Negative Bacilli].","authors":"Rıza Adaleti, Yaşar Nakipoğlu, Neslihan Arıcı, Nilgün Kansak, Şeyma Çalık, Seniha Şenbayrak, Recep Balık, Sebahat Aksaray","doi":"10.5578/mb.20239930","DOIUrl":"https://doi.org/10.5578/mb.20239930","url":null,"abstract":"<p><p>The rate of extensively drug-resistant and pan-resistant gram-negative rods isolated as infectious agents is increasing around the world and in Türkiye. One of the important options in the treatment of these infections is the combined use of antibiotics. Therefore, the aim of this study was to investigate the in vitro effect of meropenem/colistin and meropenem/fosfomycin combinations on carbapenem-resistant gram-negative bacilli isolated as infectious agents. Escherichia coli (n= 6), Klebsiella pneumoniae (n= 10), Pseudomonas aeruginosa (n= 5), and Acinetobacter baumannii (n= 6) isolates were recovered from blood and tracheal aspirate samples of patients hospitalized in our hospital's intensive care unit were included in the study. In the first stage of the combination study, minimal inhibitory concentrations (MIC) were investigated by broth microdilution for meropenem and colistin, and agar dilution methods for fosfomycin. In the second stage of the study, synergy, partial synergy, indifference, and antagonistic effects were investigated with the checkerboard method for the meropenem/colistin combination and the agar dilution method for the meropenem/fosfomycin combination. The checkerboard results were interpreted as follows: fractional inhibitory concentration index (FICI) values ≤ 0.5 synergy, < 0.5-≤ 1 partial synergy, > 1-≤ 4 indifference and FIC values of > 4 antagonism. MIC values obtained in the study were interpreted according to European Committee on Antimicrobial Susceptibility Testing (EUCAST) criteria. Of the 27 isolates studied with the broth microdilution method, 63% were found to be colistin-resistant and 37% susceptible. The MIC values of fosfomycin against Enterobacterales group bacteria were found to be in the range of 2-2048 mg/L. Two of the six E.coli isolates and nine of the 10 K.pneumoniae isolates were found to be resistant to fosfomycin (IV). The MIC values of ≥ 128 mg/L were found in all 11 non-fermentative gram-negative rods with intrinsic resistance to fosfomycin. In the combination of meropenem/ colistin, synergy and partial synergy were observed in 11 (40.7%) of 27 isolates, an indifference effect was observed in 13 (48.2%), and antagonistic effects were observed in three (11.1%) of the isolates. The synergy and partial synergy effects of this combination were 37.5% for Enterobacterales group bacteria, 50% for E.coli, and 30% for K.pneumoniae. Regarding the 11 non-fermentative gram-negative rods included in the study, 83.3% synergy and partial synergy was found in A.baumannii for the meropenem/colistin combination, while no synergy and partial synergistic effect was found in P.aeruginosa. Meropenem/fosfomycin synergy and partial synergy effects were 83.3% (5/6) for E.coli, 100% (8/8) for K.pneumoniae, 100% (6/6) for A.baumannii, and 25% (1/4) for P.aeruginosa. In all of the isolates studied, meropenem/fosfomycin combination was found to be more effective than the meropenem/colistin combination. It would be mea","PeriodicalId":18509,"journal":{"name":"Mikrobiyoloji bulteni","volume":"57 3","pages":"365-377"},"PeriodicalIF":1.0,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9886295","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Şeyda Şilan Okalın, Ayşe Nur Sarı Kaygısız, Mehmet Ali Öktem
<p><p>Numerous virulence factors are present in the hypervirulent/hypermucoviscous Klebsiella pneumoniae, which primarily causes community-acquired infections. In recent years, hypervirulent/hypermucoviscous K.pneumoniae has acquired resistance genes and has been linked to healthcare-associated infections. The aims of this study were to determine whole genome sequencing of hypermucoviscous K.pneumoniae that induces healthcare-associated bloodstream infection utilizing the Oxford Nanopore (MinION) platform, to identify resistance genes using various databases, and to compare the database results. K.pneumoniae isolates that were sent to the Dokuz Eylül University Research and Training Hospital Center Laboratory and were isolated from blood cultures were included in this study between January 2018 and December 2020. K.pneumoniae isolates were identified using the automated VITEK-2 system. The disc diffusion method was used to characterize the antimicrobial resistance profile, and the string test was used to assess the hypermucoviscous phenotype. By using specific primers blaOXA-48, blaNDM, blaKPC, blaIMP, and blaVIM, carbapenem resistance genes were investigated using the PCR method. To ascertain clonal relatedness among hypermucoviscous K.pneumoniae isolates, PFGE was used. The whole genome sequencing of five K.pneumoniae strains with different origins was determined by Oxford Nanopore (MinION) technology. Using the ResFinder, CARD and BacWGSTdb databases, resistance genes were examined. Capsule regulation genes were analyzed with the BacWGSTdb database. Resistance genes on the plasmid were discovered using the ResFinder database after plasmid analyses were carried out using the PLSDB and PlasmidFinder databases. A total of 244 K.pneumoniae isolates were included in the study. Ten isolates were found to be hypermucoviscous. A carbapenem-resistant hypermucoviscous K.pneumoniae isolate was discovered to carry the blaOXA-48. Five hypermucoviscous isolates of which whole genome sequences were determined had blaSHV types, oqxB, oqxA, and fosA genes on the chromosome. Capsule regulation genes rcsA, rcsB were detected in all five isolates, and rmpA/rmpA2 genes caused hypermucoviscous phenotype was detected in two of the five isolates. In the plasmid analysis, IncFIB (K) pCAV1099- 14, Col (pHAD28), IncFIA (HI1)-IncR, Col (pHAD28), Col440, IncFIB (pNDM_Mar)-IncHI1 (pNDM_MAR), IncL, IncFIB (K)-IncHI1 (K), IncFIB (K), IncR, IncFIB (pNDM_Mar)-IncHI1 (pNDM_MAR)-IncR, repB plasmids were identified. Resistance genes; aac(3)-IId, aph(6)-Id, aph(3")-Ib qnrB1, sul2, dfrA14, blaTEM-1A, blaCTX-M-15, armA, msr(E), mph(E), catB3, blaOXA-1, aac(6')-Ib-cr, catA1 and blaOXA-48 were detected on the plasmid. In the bioinformatic analyzes, it was determined that two study isolates with hypermucoviscous phenotype had various plasmids and carried many resistance genes on these plasmids. Various resistance and virulence genes are spread through plasmids and the number of resista
{"title":"[Determination of Whole Genome Sequence and Resistance Genes in Hypermucoviscous Klebsiella pneumoniae with Long-Read Sequencing Technology].","authors":"Şeyda Şilan Okalın, Ayşe Nur Sarı Kaygısız, Mehmet Ali Öktem","doi":"10.5578/mb.20239928","DOIUrl":"https://doi.org/10.5578/mb.20239928","url":null,"abstract":"<p><p>Numerous virulence factors are present in the hypervirulent/hypermucoviscous Klebsiella pneumoniae, which primarily causes community-acquired infections. In recent years, hypervirulent/hypermucoviscous K.pneumoniae has acquired resistance genes and has been linked to healthcare-associated infections. The aims of this study were to determine whole genome sequencing of hypermucoviscous K.pneumoniae that induces healthcare-associated bloodstream infection utilizing the Oxford Nanopore (MinION) platform, to identify resistance genes using various databases, and to compare the database results. K.pneumoniae isolates that were sent to the Dokuz Eylül University Research and Training Hospital Center Laboratory and were isolated from blood cultures were included in this study between January 2018 and December 2020. K.pneumoniae isolates were identified using the automated VITEK-2 system. The disc diffusion method was used to characterize the antimicrobial resistance profile, and the string test was used to assess the hypermucoviscous phenotype. By using specific primers blaOXA-48, blaNDM, blaKPC, blaIMP, and blaVIM, carbapenem resistance genes were investigated using the PCR method. To ascertain clonal relatedness among hypermucoviscous K.pneumoniae isolates, PFGE was used. The whole genome sequencing of five K.pneumoniae strains with different origins was determined by Oxford Nanopore (MinION) technology. Using the ResFinder, CARD and BacWGSTdb databases, resistance genes were examined. Capsule regulation genes were analyzed with the BacWGSTdb database. Resistance genes on the plasmid were discovered using the ResFinder database after plasmid analyses were carried out using the PLSDB and PlasmidFinder databases. A total of 244 K.pneumoniae isolates were included in the study. Ten isolates were found to be hypermucoviscous. A carbapenem-resistant hypermucoviscous K.pneumoniae isolate was discovered to carry the blaOXA-48. Five hypermucoviscous isolates of which whole genome sequences were determined had blaSHV types, oqxB, oqxA, and fosA genes on the chromosome. Capsule regulation genes rcsA, rcsB were detected in all five isolates, and rmpA/rmpA2 genes caused hypermucoviscous phenotype was detected in two of the five isolates. In the plasmid analysis, IncFIB (K) pCAV1099- 14, Col (pHAD28), IncFIA (HI1)-IncR, Col (pHAD28), Col440, IncFIB (pNDM_Mar)-IncHI1 (pNDM_MAR), IncL, IncFIB (K)-IncHI1 (K), IncFIB (K), IncR, IncFIB (pNDM_Mar)-IncHI1 (pNDM_MAR)-IncR, repB plasmids were identified. Resistance genes; aac(3)-IId, aph(6)-Id, aph(3\")-Ib qnrB1, sul2, dfrA14, blaTEM-1A, blaCTX-M-15, armA, msr(E), mph(E), catB3, blaOXA-1, aac(6')-Ib-cr, catA1 and blaOXA-48 were detected on the plasmid. In the bioinformatic analyzes, it was determined that two study isolates with hypermucoviscous phenotype had various plasmids and carried many resistance genes on these plasmids. Various resistance and virulence genes are spread through plasmids and the number of resista","PeriodicalId":18509,"journal":{"name":"Mikrobiyoloji bulteni","volume":"57 3","pages":"335-352"},"PeriodicalIF":1.0,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9831692","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Rukiye İnan Sarıkaya, Ömer Karaşahin, Mustafa Kemal Çoban
Brucellosis is a multisystemic infection produced by a gram-negative bacillus that can develop a variety of clinical symptoms and complications. Involvement of the central nervous system is a challenging and dangerous consequence of systemic brucellosis. The neurobrucellosis clinical spectrum can be classified as central and peripheral. Meningitis, encephalitis, polyradiculoneuritis, cranial nerve involvement, depression, abscess and cerebrovascular events are some of the potential complications that may develop. The link between neurobrucellosis and cerebrovascular accident has been reported infrequently in the literature. In this report, a case of neurobrucellosis confirmed by cerebrospinal fluid agglutination test and who developed subarachnoid hemorrhage associated with cerebral aneurysm, which is a rare condition in its course was presented. Serum Rose Bengal test and serum Brucella standard tube agglutination (STA) tests were positive at a titer of 1/640 in a 38-year-old male patient who had complaints of fever, sweating, myalgia, arthralgia, weakness, head-neck-back pain and difficulty in walking for 14 days. On magnetic resonance imaging, Brucella sacroiliitis was identified. The patient's fever, head and neck pain continued and nuchal rigidity was found to be positive. Neurobrucellosis was diagnosed based on the cerebrospinal fluid (CSF) examination, which revealed a high white blood cell count, high protein, low glucose level, and STA in CSF at 1/640 titers. Imaging of the brain was conducted concurrently with cerebrospinal fluid analysis indicated subarachnoid hemorrhage caused by cerebral aneurysm rupture. In addition to the medical treatment, the aneurysm rupture was closed with surgical intervention. Three months of simultaneous triple antibiotic treatment were administered to the patient. In the third month of the treatment, the patient was completely cured and no longer had any problems. Although uncommon, subarachnoid hemorrhage due to aneurysm rupture is one of the cerebrovascular consequences of neurobrucellosis. In the process of differential diagnosis of cerebrovascular occurrences, particularly in areas where brucellosis is an endemic disease, it is important to keep in mind that neurobrucellosis can imitate a variety of diseases and cause cerebrovascular events.
{"title":"[A Case Report on Subarachnoid Hemorrhage Secondary to Neurobrucellosis in a Patient with Cerebral Aneurysm].","authors":"Rukiye İnan Sarıkaya, Ömer Karaşahin, Mustafa Kemal Çoban","doi":"10.5578/mb.20239940","DOIUrl":"https://doi.org/10.5578/mb.20239940","url":null,"abstract":"<p><p>Brucellosis is a multisystemic infection produced by a gram-negative bacillus that can develop a variety of clinical symptoms and complications. Involvement of the central nervous system is a challenging and dangerous consequence of systemic brucellosis. The neurobrucellosis clinical spectrum can be classified as central and peripheral. Meningitis, encephalitis, polyradiculoneuritis, cranial nerve involvement, depression, abscess and cerebrovascular events are some of the potential complications that may develop. The link between neurobrucellosis and cerebrovascular accident has been reported infrequently in the literature. In this report, a case of neurobrucellosis confirmed by cerebrospinal fluid agglutination test and who developed subarachnoid hemorrhage associated with cerebral aneurysm, which is a rare condition in its course was presented. Serum Rose Bengal test and serum Brucella standard tube agglutination (STA) tests were positive at a titer of 1/640 in a 38-year-old male patient who had complaints of fever, sweating, myalgia, arthralgia, weakness, head-neck-back pain and difficulty in walking for 14 days. On magnetic resonance imaging, Brucella sacroiliitis was identified. The patient's fever, head and neck pain continued and nuchal rigidity was found to be positive. Neurobrucellosis was diagnosed based on the cerebrospinal fluid (CSF) examination, which revealed a high white blood cell count, high protein, low glucose level, and STA in CSF at 1/640 titers. Imaging of the brain was conducted concurrently with cerebrospinal fluid analysis indicated subarachnoid hemorrhage caused by cerebral aneurysm rupture. In addition to the medical treatment, the aneurysm rupture was closed with surgical intervention. Three months of simultaneous triple antibiotic treatment were administered to the patient. In the third month of the treatment, the patient was completely cured and no longer had any problems. Although uncommon, subarachnoid hemorrhage due to aneurysm rupture is one of the cerebrovascular consequences of neurobrucellosis. In the process of differential diagnosis of cerebrovascular occurrences, particularly in areas where brucellosis is an endemic disease, it is important to keep in mind that neurobrucellosis can imitate a variety of diseases and cause cerebrovascular events.</p>","PeriodicalId":18509,"journal":{"name":"Mikrobiyoloji bulteni","volume":"57 3","pages":"481-489"},"PeriodicalIF":1.0,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9831697","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
<p><p>Crimean-Congo hemorrhagic fever (CCHF) is an acute febrile hemorrhagic disease that can be fatal. Almost one-eighth of people infected with CCHF develop serious illness. The mortality rate is high due to severe bleeding, diffuse intravascular coagulation, shock, and multiple organ failure. Early detection of serious illness can play a key role in developing effective treatment and follow-up strategies. C-reactive protein (CRP), blood urea nitrogen (BUN), and albumin have previously been evaluated as markers of clinical severity in infectious diseases. This study aimed to evaluate the role of these readily available and inexpensive biomarkers and their ratios as predictors of mortality risk in patients with CCHF. This retrospective observational single-center study was conducted between May and October 2022 in a regional hospital in northeastern Türkiye, where the incidence of CCHF is the highest. Hundred and fifty patients aged 18 years and over with a definitive diagnosis of CCHF were included; patients with chronic kidney disease requiring long-term hemodialysis and those with missing data were excluded from the study. The patients' demographic characteristics, comorbidities, initial complaints, and epidemiological, clinical, and laboratory findings were recorded. Receiver operating characteristics (ROC) curve analysis was used to determine the predictive power of the studied biomarkers. Categorical and continuous variables found to be significant for mortality were evaluated using univariate logistic regression. Variables found to be significant in this test were used to create a multivariate logistic regression model to identify independent risk factors for mortality. The median age of the patients was 49 (18-89) years and 93 (62.0%) were men. Twelve patients (8.0%) required intensive care and 11 (7.3%) died. Complaints of abdominal pain (p= 0.010), hypotension (p= 0.002), somnolence (p< 0.001), and bleeding (p< 0.001) at the time of hospital admission were significantly more common among non-surviving patients. BUN and CRP were the biomarkers with the highest diagnostic power for mortality. A BUN cut-off value of 19.5 mg/dl had 100% sensitivity and 74.1% specificity, while a CRP cut-off value of 31.5 mg/L had 100% sensitivity and 81.8% specificity. CRP/albumin ratio (CAR) and BUN/albumin ratio (BAR) had higher predictive power than all individual biomarkers. At a cut-off point of 0.98, CAR had diagnostic power of 0.942 (95% confidence interval= 0.901-0.984), 100% sensitivity, and 84.9% specificity for mortality. At a cut-off of 0.50, BAR predicted mortality with diagnostic power of 0.932 (95% confidence interval= 0.879-0.984), 100% sensitivity, and 81.3% specificity. In univariate logistic regression analysis, the presence of bleeding, somnolence, and hypotension at the time of admission; higher troponin, total bilirubin, neutrophil count, activated partial thromboplastin time, prothrombin time, and age; and lower platelet count, fibri
{"title":"[The Role of Blood Urea Nitrogen and C-Reactive Protein and Their Ratios to Albumin in Predicting Mortality in Crimean-Congo Hemorrhagic Fever].","authors":"Ömer Karaşahin, Rukiye İnan Sarıkaya","doi":"10.5578/mb.20239934","DOIUrl":"https://doi.org/10.5578/mb.20239934","url":null,"abstract":"<p><p>Crimean-Congo hemorrhagic fever (CCHF) is an acute febrile hemorrhagic disease that can be fatal. Almost one-eighth of people infected with CCHF develop serious illness. The mortality rate is high due to severe bleeding, diffuse intravascular coagulation, shock, and multiple organ failure. Early detection of serious illness can play a key role in developing effective treatment and follow-up strategies. C-reactive protein (CRP), blood urea nitrogen (BUN), and albumin have previously been evaluated as markers of clinical severity in infectious diseases. This study aimed to evaluate the role of these readily available and inexpensive biomarkers and their ratios as predictors of mortality risk in patients with CCHF. This retrospective observational single-center study was conducted between May and October 2022 in a regional hospital in northeastern Türkiye, where the incidence of CCHF is the highest. Hundred and fifty patients aged 18 years and over with a definitive diagnosis of CCHF were included; patients with chronic kidney disease requiring long-term hemodialysis and those with missing data were excluded from the study. The patients' demographic characteristics, comorbidities, initial complaints, and epidemiological, clinical, and laboratory findings were recorded. Receiver operating characteristics (ROC) curve analysis was used to determine the predictive power of the studied biomarkers. Categorical and continuous variables found to be significant for mortality were evaluated using univariate logistic regression. Variables found to be significant in this test were used to create a multivariate logistic regression model to identify independent risk factors for mortality. The median age of the patients was 49 (18-89) years and 93 (62.0%) were men. Twelve patients (8.0%) required intensive care and 11 (7.3%) died. Complaints of abdominal pain (p= 0.010), hypotension (p= 0.002), somnolence (p< 0.001), and bleeding (p< 0.001) at the time of hospital admission were significantly more common among non-surviving patients. BUN and CRP were the biomarkers with the highest diagnostic power for mortality. A BUN cut-off value of 19.5 mg/dl had 100% sensitivity and 74.1% specificity, while a CRP cut-off value of 31.5 mg/L had 100% sensitivity and 81.8% specificity. CRP/albumin ratio (CAR) and BUN/albumin ratio (BAR) had higher predictive power than all individual biomarkers. At a cut-off point of 0.98, CAR had diagnostic power of 0.942 (95% confidence interval= 0.901-0.984), 100% sensitivity, and 84.9% specificity for mortality. At a cut-off of 0.50, BAR predicted mortality with diagnostic power of 0.932 (95% confidence interval= 0.879-0.984), 100% sensitivity, and 81.3% specificity. In univariate logistic regression analysis, the presence of bleeding, somnolence, and hypotension at the time of admission; higher troponin, total bilirubin, neutrophil count, activated partial thromboplastin time, prothrombin time, and age; and lower platelet count, fibri","PeriodicalId":18509,"journal":{"name":"Mikrobiyoloji bulteni","volume":"57 3","pages":"419-431"},"PeriodicalIF":1.0,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9862397","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
<p><p>The aim of this study was to investigate the frequency of Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis and Mycoplasma genitalium in men living with HIV in terms of sociodemographic characteristics and behavioral risk factors. In this cross-sectional, single center study, all HIV-infected male patients, aged ≥ 18 years, including those being followed-up (n= 142) and the new admissions (n= 16) at Hacettepe University, Department of Infectious Diseases between March 1st, 2017 and May 1st, 2018 were included. After obtaining the informed consent form; age, follow-up days in STI-clinic, marital status, education, employment status; STI-related sign and symptoms, prior STI diagnosis, multiple sexual partners during the last year, exchanging sex for money, sexual orientation, drug use, condom use with regular and casual partner and also risk factors regarding partners were inquired as behavioural risk factors. A sample of first-voided urine of each participant was tested for the presence of Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis and Mycoplasma genitalium by using nucleic acid amplification test (NAAT) (BD-MAX system, BD Diagnostics, USA) and BD MAX Mycoplasma-Ureaplasma-OSR for BioGX, (BD Diagnostics, The Netherlands). All participants living with HIV, men who have sex with men (MSM) and heterosexual men were grouped as STI-positive and STI-negative and compared. For all statistical analysis, SPSS 24 software was used. During the period of 14 months; the data was determined as follows: median follow-up time was 1138 (IQR= 159.5- 1494.5) days, median age was 35 (IQR= 28-42) years, 73.3% were single, 68.3% were at least college graduates or had higher educational attainment, 78.1% were currently employed. Of the participants, 26.9% reported STI-related sign and symptoms, 50.0% at least one STI episode in the past. Nine (5.6%) M.genitalium, five (3.1%) N.gonorrhoeae, and four (2.5%) C.trachomatis were detected in the urine samples of 17 (10.7%) individuals. N.gonorrhoeae and C.trachomatis were detected simultaneously in only one patient's urine sample. STI-positive patients (n= 17) were determined to be younger compared to STI-negative group [(p= 0.02; 27 years (IQR= 24-37) vs 35 years (IQR= 28-42)], had prominent STI-related signs and symptoms (p< 0.001) and had more multiple sexual partners (p= 0.03). The median CD4+ T lymphocyte count were relatively lower (p= 0.03) in STI-positive patients and plasma HIV RNA level was higher compared to the STI-negative participants (p= 0.05). STI-positive MSM group were younger [p= 0.01; 26 years (IQR= 23.5-29) vs 33 years, (IQR= 28-40)], STI-related signs and symptoms were more prominent (p= 0.02), the frequency of exchanging sex for money/drugs among their partners (p= 0.03) was higher compared to their STI-negative counterparts. Among STI-positive heterosexual patients, the presence of STI-related signs and symptoms (p= 0.04), drug use among their partners (
{"title":"[Investigation of the Frequency of Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis and Mycoplasma genitalium in Men Living with HIV in Terms of Sociodemographic Characteristics and Behavioral Risk Factors].","authors":"Çağlayan Merve Ayaz, Nesrin Damla Karakaplan, Ahmet Çağkan İnkaya, Banu Çakır, Serhat Ünal, Pınar Zarakolu","doi":"10.5578/mb.20239931","DOIUrl":"https://doi.org/10.5578/mb.20239931","url":null,"abstract":"<p><p>The aim of this study was to investigate the frequency of Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis and Mycoplasma genitalium in men living with HIV in terms of sociodemographic characteristics and behavioral risk factors. In this cross-sectional, single center study, all HIV-infected male patients, aged ≥ 18 years, including those being followed-up (n= 142) and the new admissions (n= 16) at Hacettepe University, Department of Infectious Diseases between March 1st, 2017 and May 1st, 2018 were included. After obtaining the informed consent form; age, follow-up days in STI-clinic, marital status, education, employment status; STI-related sign and symptoms, prior STI diagnosis, multiple sexual partners during the last year, exchanging sex for money, sexual orientation, drug use, condom use with regular and casual partner and also risk factors regarding partners were inquired as behavioural risk factors. A sample of first-voided urine of each participant was tested for the presence of Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis and Mycoplasma genitalium by using nucleic acid amplification test (NAAT) (BD-MAX system, BD Diagnostics, USA) and BD MAX Mycoplasma-Ureaplasma-OSR for BioGX, (BD Diagnostics, The Netherlands). All participants living with HIV, men who have sex with men (MSM) and heterosexual men were grouped as STI-positive and STI-negative and compared. For all statistical analysis, SPSS 24 software was used. During the period of 14 months; the data was determined as follows: median follow-up time was 1138 (IQR= 159.5- 1494.5) days, median age was 35 (IQR= 28-42) years, 73.3% were single, 68.3% were at least college graduates or had higher educational attainment, 78.1% were currently employed. Of the participants, 26.9% reported STI-related sign and symptoms, 50.0% at least one STI episode in the past. Nine (5.6%) M.genitalium, five (3.1%) N.gonorrhoeae, and four (2.5%) C.trachomatis were detected in the urine samples of 17 (10.7%) individuals. N.gonorrhoeae and C.trachomatis were detected simultaneously in only one patient's urine sample. STI-positive patients (n= 17) were determined to be younger compared to STI-negative group [(p= 0.02; 27 years (IQR= 24-37) vs 35 years (IQR= 28-42)], had prominent STI-related signs and symptoms (p< 0.001) and had more multiple sexual partners (p= 0.03). The median CD4+ T lymphocyte count were relatively lower (p= 0.03) in STI-positive patients and plasma HIV RNA level was higher compared to the STI-negative participants (p= 0.05). STI-positive MSM group were younger [p= 0.01; 26 years (IQR= 23.5-29) vs 33 years, (IQR= 28-40)], STI-related signs and symptoms were more prominent (p= 0.02), the frequency of exchanging sex for money/drugs among their partners (p= 0.03) was higher compared to their STI-negative counterparts. Among STI-positive heterosexual patients, the presence of STI-related signs and symptoms (p= 0.04), drug use among their partners (","PeriodicalId":18509,"journal":{"name":"Mikrobiyoloji bulteni","volume":"57 3","pages":"378-389"},"PeriodicalIF":1.0,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9886294","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Didem Özgür, Leyla Ersoy, Mahmut Ülger, Seda Tezcan Ülger, Gönül Aslan
<p><p>Tuberculosis (TB) is caused by Mycobacterium tuberculosis, still one of the most common life-threatening infectious diseases worldwide. Although drug resistance in M.tuberculosis is mainly due to spontaneous chromosomal mutations in genes encoding drug target or drug activating enzymes, the resistance cannot be explained only by these mutations. Low permeability of the cell wall, drug inactivating enzymes and especially efflux pumps (EPs) are other mechanisms of drug resistance in mycobacteria. Efflux pump inhibitors (EPIs) binding to M.tuberculosis EPs were shown to inhibit efflux of anti-TB drugs, to enhance M.tuberculosis killing, to reduce drug resistance and to produce synergistic effects with first line anti-TB drugs. In this study, we aimed to determine the minimum inhibitory concentration (MIC) of first-line anti-TB drugs in the presence of verapamil (VER) and the expression of 21 putative EP genes belonged to the ATP-binding cassette (ABC), major facilitator superfamily (MFS) and resistance-nodulation-division (RND) families which might have caused the resistance in nine M.tuberculosis complex clinical isolates resistant to all of the first line anti-TB drugs. MIC values of the isolates were determined in 96-well U-bottom plates by the resazurin microtiter test (REMA) method based on the color change principle. According to the determined MIC values of each isolate, freshly grown cultures in Middlebrook 7H9 broth were exposed to first-line anti-TB drugs and MIC of first-line anti-TB drugs in the presence of VER (½ MIC) at 37°C for 48 hours for RNA extraction. The non-drug exposed cultures were used as control. Total RNA was extracted using the RNeasy Mini Kit (Qiagen GmbH, Hilden, Germany) and then treated with DNase I (Thermo Fischer Scientific Inc., Waltham, MA). Complementary DNA (cDNA) from the extracted RNAs was synthesized with the "First strand cDNA synthesis kit" (Thermo Fischer Scientific Inc., Waltham, MA) using oligo primers. The expression levels of efflux pump genes by quantitative realtime polymerase chain reaction (qRt-PCR) were performed using the QuantiTect SYBR Green Rt-PCR Kit (Qiagen, Germany). The housekeeping sigma factor gene sigA (Rv2703) was used as internal control in qRt‑PCR assays. Relative quantification of the clinical isolates was determined by the 2-∆∆Ct method by comparing the expression levels of efflux genes in cultures exposed to primary anti-TB drugs and VER with those of non-drug exposed cultures. MIC values of nine isolates by REMA method was determined between 32-512 μg/mL, 1-128 μg/mL, 2-32 μg/mL, 4-16 μg/mL and 15.62-250 μg/mL for streptomycin (SM), isoniazid (INH), rifampicin (RIF), ethambutol (EMB) and VER, respectively. In the presence of ½ MIC VER, it was determined that the MIC of SM decreased 2-32 fold in eight isolates, the MIC of INH decreased by 2-8 fold in nine isolates, the MIC of RIF decreased by 2-16 fold in eight isolates, and the MIC of EMB decreased 2-4 fold in only five iso
{"title":"[Investigation of Efflux Pump Genes in Resistant Mycobacterium tuberculosis Complex Clinical Isolates Exposed to First Line Antituberculosis Drugs and Verapamil Combination].","authors":"Didem Özgür, Leyla Ersoy, Mahmut Ülger, Seda Tezcan Ülger, Gönül Aslan","doi":"10.5578/mb.20239916","DOIUrl":"https://doi.org/10.5578/mb.20239916","url":null,"abstract":"<p><p>Tuberculosis (TB) is caused by Mycobacterium tuberculosis, still one of the most common life-threatening infectious diseases worldwide. Although drug resistance in M.tuberculosis is mainly due to spontaneous chromosomal mutations in genes encoding drug target or drug activating enzymes, the resistance cannot be explained only by these mutations. Low permeability of the cell wall, drug inactivating enzymes and especially efflux pumps (EPs) are other mechanisms of drug resistance in mycobacteria. Efflux pump inhibitors (EPIs) binding to M.tuberculosis EPs were shown to inhibit efflux of anti-TB drugs, to enhance M.tuberculosis killing, to reduce drug resistance and to produce synergistic effects with first line anti-TB drugs. In this study, we aimed to determine the minimum inhibitory concentration (MIC) of first-line anti-TB drugs in the presence of verapamil (VER) and the expression of 21 putative EP genes belonged to the ATP-binding cassette (ABC), major facilitator superfamily (MFS) and resistance-nodulation-division (RND) families which might have caused the resistance in nine M.tuberculosis complex clinical isolates resistant to all of the first line anti-TB drugs. MIC values of the isolates were determined in 96-well U-bottom plates by the resazurin microtiter test (REMA) method based on the color change principle. According to the determined MIC values of each isolate, freshly grown cultures in Middlebrook 7H9 broth were exposed to first-line anti-TB drugs and MIC of first-line anti-TB drugs in the presence of VER (½ MIC) at 37°C for 48 hours for RNA extraction. The non-drug exposed cultures were used as control. Total RNA was extracted using the RNeasy Mini Kit (Qiagen GmbH, Hilden, Germany) and then treated with DNase I (Thermo Fischer Scientific Inc., Waltham, MA). Complementary DNA (cDNA) from the extracted RNAs was synthesized with the \"First strand cDNA synthesis kit\" (Thermo Fischer Scientific Inc., Waltham, MA) using oligo primers. The expression levels of efflux pump genes by quantitative realtime polymerase chain reaction (qRt-PCR) were performed using the QuantiTect SYBR Green Rt-PCR Kit (Qiagen, Germany). The housekeeping sigma factor gene sigA (Rv2703) was used as internal control in qRt‑PCR assays. Relative quantification of the clinical isolates was determined by the 2-∆∆Ct method by comparing the expression levels of efflux genes in cultures exposed to primary anti-TB drugs and VER with those of non-drug exposed cultures. MIC values of nine isolates by REMA method was determined between 32-512 μg/mL, 1-128 μg/mL, 2-32 μg/mL, 4-16 μg/mL and 15.62-250 μg/mL for streptomycin (SM), isoniazid (INH), rifampicin (RIF), ethambutol (EMB) and VER, respectively. In the presence of ½ MIC VER, it was determined that the MIC of SM decreased 2-32 fold in eight isolates, the MIC of INH decreased by 2-8 fold in nine isolates, the MIC of RIF decreased by 2-16 fold in eight isolates, and the MIC of EMB decreased 2-4 fold in only five iso","PeriodicalId":18509,"journal":{"name":"Mikrobiyoloji bulteni","volume":"57 2","pages":"207-219"},"PeriodicalIF":1.0,"publicationDate":"2023-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9687958","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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