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Volume 7,
Issue 5,
2025
Volume 7, Issue 5, 2025
- Research Articles
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Is the Zika virus re-emerging as a distinct genetic lineage in India?
show More to view fulltext, buy and share links for: show Less to hide fulltext, buy and share links for:An outbreak of Zika fever occurred in Thiruvananthapuram City, Kerala, India, during 2021. At the request of the Kerala state health administration, we investigated the same, towards proposing requisite containment strategies for the disease outbreak. Epidemiological investigations indicated a clustering pattern of Zika fever cases with the presumed index case from a multi-speciality hospital in the city. Preliminary reports on the same had been already reported elsewhere during 2021. Further, entomological surveys carried out evinced the predominant mosquito species in the city, viz. Aedes albopictus (65.55%), Aedes aegypti (22.0%) and Aedes vittatus (12.0%) were naturally infected with Zika virus (ZIKV), the minimum infection rates being 17.9, 7.8 and 3.6, respectively. Also, trans-ovarian transmission was recorded in both Ae. aegypti and Ae. albopictus. This is the first report on the detection of ZIKV from Ae. albopictus in India. Analysis of phylogenetically informative genes of the ZIKV genome indicated the emergence of a distinct lineage of the Asian strain of virus, with five unique non-synonymous mutations, viz. ‘A22T’ and ‘I160M’ (pre-membrane) and ‘D348N’, ‘T470A’ and ‘V473L’ (envelope), that were involved in the outbreak. The altered gene expression pattern and evolutionary implications of these unique mutations remain to be investigated. Genetic analysis of the virus isolates from this and other investigations carried out on sporadic outbreaks of ZIKV in the country subsequently indicated that ZIKV is re-emerging as a distinct genetic lineage in India. These findings and other recent reports on ZIKV outbreaks warrant an urgent need for a systematic countrywide surveillance strategy, towards the prevention/preparedness/containment of a massive outbreak of this emerging neurovirulent arboviral disease.
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Haematological abnormalities as diagnostic indicators of malaria in returning travellers: a retrospective study at Mohamed V Military Instruction Hospital
show More to view fulltext, buy and share links for: show Less to hide fulltext, buy and share links for:Introduction. Malaria remains a significant global health concern, particularly in travellers returning from endemic regions. Haematological abnormalities are often associated with malaria and can serve as diagnostic indicators, especially when clinical symptoms are nonspecific.
Objective. This study aims to identify the most relevant haematological parameters for diagnosing malaria in travellers returning from endemic areas, who sought care at the Mohamed V Military Instruction Hospital in Rabat.
Methods. We conducted a retrospective comparative study involving 829 patients who returned from malaria-endemic regions between January 2017 and December 2023. Data collected included demographic information, parasitological test results and comprehensive haematological profiles. Statistical analysis was performed to determine the sensitivity and specificity of various haematological parameters in diagnosing malaria.
Results. Thrombocytopenia, lymphocytopenia and anaemia were the most significant haematological abnormalities associated with malaria. Thrombocytopenia, defined as a platelet count below 150×10³ µl−1, demonstrated a sensitivity of 75.91% and a specificity of 84.11%. Lymphocytopenia, with a threshold of less than 1.5×10³ µl−1, showed a sensitivity of 69.47% and a specificity of 78.39%. Anaemia, defined by haemoglobin levels below 13 g dl−1 in men and 12 g dl−1 in women, also significantly correlated with malaria diagnosis.
Discussion. This study highlights the significance of haematological abnormalities as key diagnostic markers for imported malaria cases. By analysing retrospective data, we observed that these abnormalities, especially thrombocytopenia and anaemia, are common amongst returning travellers with confirmed malaria. These findings suggest that clinicians can use such markers as a valuable tool for early malaria diagnosis, potentially improving patient outcomes. Additionally, the study reinforces the need for heightened awareness amongst healthcare providers in non-endemic regions regarding the presentation of malaria in travellers.
Conclusion. Haematological parameters such as thrombocytopenia, lymphocytopenia and anaemia are valuable diagnostic tools for malaria in returning travellers. These findings suggest that these parameters should be integrated into diagnostic protocols to improve the accuracy and timeliness of malaria diagnosis, particularly in clinical settings with limited access to advanced diagnostic tools.
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Detection and antibiotic resistance of Salmonella isolates from selected poultry farms in Dar es Salaam, Tanzania
show More to view fulltext, buy and share links for: show Less to hide fulltext, buy and share links for:Introduction. Salmonella is one of the most prevalent foodborne bacteria, posing a significant global health concern and responsible for ~155,000 deaths and 93.8 million human foodborne illnesses annually. The rampant use of antibiotic agents to combat salmonellosis in poultry has contributed to the emergence of resistance against commonly used antibiotics.
Methodology. The cross-sectional study was conducted between January and June 2023 across three districts in the Dar es Salaam region, Tanzania. Salmonella isolates were detected and confirmed by using standard microbiological conventional methods and molecular methods such as PCR and genomic sequencing. PCR was used for detecting the presence of the invA gene, and partial DNA sequencing was performed to identify species and their close relatedness. A Kirby–Bauer disc diffusion method was employed to evaluate Salmonella sensitivity to seven different commonly used antibiotics, namely ampicillin, azithromycin, chloramphenicol, ciprofloxacin, gentamycin, trimethoprim/sulfamethoxazole and tetracycline. Salmonella strain with reference number ATCC 8739 was used as a control.
Results. The overall Salmonella isolates from faecal droppings were 6.04% (n=796). Of the PCR-confirmed isolates, 64.3% (n=28) were resistant to more than two classes of antibiotics and hence considered multidrug resistant. The highest resistance was observed with ampicillin (92.9%, n=26), followed by tetracycline (71.43%, n=20), ciprofloxacin (42.9%, n=12), sulphonamide (42.85%, n=12), gentamicin (35.7%, n=10) and azithromycin (28.9%, n=8). All the isolates were susceptible to chloramphenicol (100%, n=28). Twenty-eight isolates were sent for sequencing, out of which 16 sequences (OR021717-OR021739) met the criteria for phylogenetic analysis. All 16 sequences had a per cent identity to EU348369 strains Senftenberg isolated from China, OL581594 Salmonella Newport isolated from China and EU348368 Salmonella Pullorum isolates from China. Other sequences diverged more distantly; these are Salmonella Abony with accession number CP007541 and Salmonella Kentucky with accession number OL581592. The tree also included an outgroup species, Salmonella bongori, which was downloaded from GenBank with accession numbers NC015761 and NC021870 S. bongori.
Conclusion. The high level of antibiotic resistance found in this study could be due to the misuse of antibiotics in poultry management, and/or, probably, there were circulating resistant Salmonella strains in the environment. To reverse the trend observed, immediate interventions such as advocating for the prudent use of antibiotics in poultry production systems by strengthening extension services to poultry farmers and the use of a farmer field school model to improve poultry management through improved farm biosecurity are required.
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Derivatization of pBACpAK entrapment vectors for enhanced mobile genetic element transposition detection in multidrug-resistant Escherichia coli
show More to view fulltext, buy and share links for: show Less to hide fulltext, buy and share links for:Aim. Antimicrobial resistance poses a critical global health threat, driven by the dissemination of resistance genes via mobile genetic elements (MGEs). This study aims to enhance the detection of MGE insertions in multidrug-resistant Escherichia coli by derivatizing the pBACpAK entrapment vector.
Methods and results. Three derivatives were constructed with additional nucleotides upstream of the cI repressor gene, based on conserved regions identified from GenBank sequences containing known IS26 and IS1 insertions. Using colony PCR, intracellular transposition screening was performed on 194 tetracycline-resistant colonies from four E. coli ESI123 strains carrying different pBACpAK constructs. The derivatives showed increased MGE capture rates (10.7–73.1 %) compared to the WT vector (3.75%), identifying multiple MGEs, including the novel composite transposon Tn7824. Tn7824 harbours the bla OXA-181 carbapenem resistance gene and the qnrS1 quinolone resistance gene, highlighting the clinical relevance of these findings. Long-read sequencing of transposants confirmed the accuracy of MGE identification and structural characterization, which also revealed chromosomal integration events of the pBACpAK derivatives mediated by flanking insertion sequences.
Conclusions. The modifications introduced in the pBACpAK derivatives could increase the detection of transposition events by alleviating spatial constraints, allowing for more robust MGE detection.
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Multi-metal-resistant Staphylococcus warneri strain TWSL_1: revealing heavy metal-resistant genomic features by whole-genome sequencing and analysis
show More to view fulltext, buy and share links for: show Less to hide fulltext, buy and share links for:This study explores the genomic basis of heavy metal resistance in Staphylococcus warneri strain TWSL_1, isolated from industrial textile effluent. The strain exhibited strong resistance to Cd²+, Pb²+ and Cu²+, with MICs of 50, 1,200 and 75 mg l−1, respectively. Whole-genome sequencing revealed a 2.66 Mb genome with 2,567 CDSs and a 99.81% average nucleotide identity to S. warneri WS479. Comparative genomic analysis at the genus level revealed that S. warneri strain TWSL_1 possesses a unique and expanded repertoire of heavy metal resistance genes, including the cadmium efflux system accessory protein and cadmium resistance protein, which are absent in pathogenic Staphylococcus sp. used for the comparison. Phylogenetic analysis confirmed its classification within S. warneri, with strong bootstrap support (100). Functional annotation highlighted metabolic versatility and stress response capabilities, supporting its adaptation to metal-rich environments. S. warneri TWSL_1 demonstrated high Pb²+ removal efficiency, reducing concentrations by over 70%. These findings highlight S. warneri TWSL_1 as a promising candidate for heavy metal bioremediation with potential applications in environmental detoxification and monitoring strategies.
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Are reads required? High-precision variant calling from bacterial genome assemblies
show More to view fulltext, buy and share links for: show Less to hide fulltext, buy and share links for:Accurate nucleotide variant calling is essential in microbial genomics, particularly for outbreak tracking and phylogenetics. This study evaluates variant calls derived from genome assemblies compared to traditional read-based variant-calling methods, using seven closely related Staphylococcus aureus isolates sequenced on Illumina and Oxford Nanopore Technologies platforms. By benchmarking multiple assembly and variant-calling pipelines against a ground truth dataset, we found that read-based methods consistently achieved high accuracy. Assembly-based approaches performed well in some cases but were highly dependent on assembly quality, as errors in the assembly led to false-positive variant calls. These findings underscore the need for improved assembly techniques before the potential benefits of assembly-based variant calling (such as reduced computational requirements and simpler data management) can be realized.
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- Short Communications
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Classification of unsequenced Mycobacterium tuberculosis strains in a high-burden setting using a pairwise logistic regression approach
show More to view fulltext, buy and share links for: show Less to hide fulltext, buy and share links for:Over the past three decades, molecular epidemiological studies have provided new opportunities to investigate the transmission dynamics of Mycobacterium tuberculosis. In most studies, a sizable fraction of individuals with notified tuberculosis cannot be included, either because they do not have culture-positive disease (and thus do not have specimens available for molecular typing) or because resources for conducting sequencing are limited. A recent study introduced a regression-based approach for inferring the membership of unsequenced tuberculosis cases in transmission clusters based on host demographic and epidemiological data. This method was able to identify the most likely cluster to which an unsequenced strain belonged with an accuracy of 35%, although this was in a low-burden setting where a large fraction of cases occurred among foreign-born migrants. Here, we apply a similar model to M. tuberculosis whole-genome sequencing data from the Republic of Moldova, a setting of relatively high local transmission. Using a maximum cluster span of ~40 single nucleotide polymorphisms (SNPs) and a cluster size cutoff of n≥10, we could best predict the specific cluster to which each clustered case was most likely to be a member with an accuracy of 17.2 %. In sensitivity analyses, we found that a more restrictive (~20 SNPs threshold) or permissive (~80 SNPs) threshold did not improve performance. We found that increasing the minimum cluster size improved prediction accuracy. These findings highlight the challenges of transmission inference in high-burden settings like Moldova.
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Draft genome sequences of four potential new species of the genus Bradyrhizobium isolated from root nodules of native legumes in Costa Rican forests
show More to view fulltext, buy and share links for: show Less to hide fulltext, buy and share links for:Here, we report the draft genome sequences of four Bradyrhizobium spp. isolates obtained from root nodules of the native legumes Pentaclethra macroloba, Chamaecrista nictitans, Erythrina fusca and Zygia engelsingii in tropical forests of Costa Rica. Genomes ranged from 8.6 to 9.8 Mb with GC contents between 62.8% and 63.8%. Phylogenomic analysis, along with average nucleotide identity (ANI) and digital DNA–DNA hybridization (dDDH) comparisons, confirmed that these isolates represent potential new species. ANI values ranged from 88.3% to 90.3%, and dDDH values from 28.8% to 41.8%, compared with their closest Bradyrhizobium species. Functional annotation revealed some genes related to nitrogen fixation (nifA, nifB, nifH) and nodulation capacity (nodB, nodC, nodJ). These results provide insights into the diversity and symbiotic capabilities of Bradyrhizobium in tropical ecosystems.
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- Case Reports
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Cardiac tamponade revealing tuberculous pericarditis: a case report
show More to view fulltext, buy and share links for: show Less to hide fulltext, buy and share links for:Tuberculosis remains a major public health issue in Morocco. Pulmonary tuberculosis is the most common form, but various extrapulmonary forms exist. Tuberculous pericarditis is a rare form of extrapulmonary tuberculosis that can be complicated by cardiac tamponade, pericardial constriction or their combination, which can threaten the patient’s life. Its clinical and radiological signs are nonspecific, and the clinical presentation can be misleading and incomplete, sometimes even with an initial tamponade. We report the case of a 68-year-old female patient admitted for intense retrosternal chest pain associated with acute dyspnoea, evolving in the context of unquantified weight loss and general deterioration. Additionally, she reported a history of fever and night sweats. Clinical examination revealed a conscious, febrile, hypotensive, tachycardic, polypneic patient with good oxygen saturation, signs of right heart failure and muffled heart sounds on auscultation. Chest X-ray revealed cardiomegaly, and the ECG showed diffuse low voltage. Given the presence of Beck’s triad suggestive of cardiac tamponade, a transthoracic echocardiogram was performed, revealing a large pericardial effusion with a ‘swinging heart’. A chest CT scan also confirmed the large pericardial effusion. The diagnosis of cardiac tamponade was made based on the clinical and radiological findings, and pericardial drainage was performed, after which the patient showed clinical improvement. PCR GenXpert MTB/RIF Ultra detected the presence of Mycobacterium tuberculosis in the pericardial fluid, with no resistance to rifampicin. Culture was positive for M. tuberculosis. The diagnosis of tuberculous pericarditis was, thus, confirmed, and the patient was started on quadruple antituberculosis therapy with good clinical progress.
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Primary iliac bone tuberculosis: a case report
show More to view fulltext, buy and share links for: show Less to hide fulltext, buy and share links for:Tuberculosis (TB) remains one of the world’s leading causes of morbidity and mortality. It occurs in both pulmonary and extra-pulmonary forms. Primary iliac bone TB remains a rare clinical entity, even in endemic areas. The diagnosis of the disease can be challenging due to its similarity to other bone diseases. We report a rare case of primary iliac bone TB in a 63-year-old patient who was on peritoneal dialysis and had a medical history of hypertension and type II diabetes, which was complicated by diabetic retinopathy and diabetic kidney disease. Magnetic resonance imaging revealed osteomyelitis in the iliac bone, while real-time polymerase chain reaction using the GeneXpert® system on a gluteal collection sample confirmed the diagnosis of TB. The integration of advanced molecular tools, such as GeneXpert®, represents significant progress, enabling rapid and accurate diagnosis of TB and facilitating early initiation of treatment.
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Gummatous mitral valve endocarditis from tertiary syphilis
show More to view fulltext, buy and share links for: show Less to hide fulltext, buy and share links for:A 50-year-old Romanian gentleman presented with fever, myalgia and 30 kg weight loss. He was treated for syphilis after acquiring it 16 years ago. On examination, there was a pansystolic murmur in the axilla, and the patient had an ataxic gait. Blood tests showed raised inflammatory markers. However, standard investigations for infective endocarditis, including multiple blood cultures, serological titres for fastidious organisms and antibody tests were negative. A computed tomography (CT) of the chest, abdomen and pelvis demonstrated hepatosplenomegaly with multiple splenic infarcts. A magnetic resonance imaging (MRI) of the head with contrast showed multiple punctate enhancement in the bilateral hemispheres with leptomeningeal enhancement. Transthoracic echocardiogram demonstrated a large vegetation leading to severe mitral regurgitation. Serum treponemal antibodies were positive; Treponema pallidum particle agglutination (TPPA) was positive at 1 : 1280, and rapid plasma reagin (RPR) 1 : 4 treponemal IgM was negative; lumbar puncture syphilis serology was negative. The patient was treated with an extensive period of intravenous antibiotics, in addition to a prosthetic metallic valve replacement, where unusual ragged calcified valvular tissue was observed. Tertiary syphilis is a difficult diagnosis to confirm, since it can often be indolent and occur in areas of the body where it may go unnoticed. In our case, a diagnosis of probable syphilitic endocarditis was made from a combination of the history, an initial increase in the size of the lesion following antibiotic therapy and observation of likely gumma on the mitral valve during surgery. In such cases, surgery in addition to optimal antimicrobial therapy is necessary for effective treatment. This case adds to the current literature that treatment with penicillin is likely inadequate to prevent late complications.
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First case of infective endocarditis due to NDM-type carbapenemase-producing Serratia marcescens in a preterm infant: a case report
show More to view fulltext, buy and share links for: show Less to hide fulltext, buy and share links for:Serratia marcescens (S. marcescens) is a Gram-negative rod-shaped bacterium belonging to the Enterobacteriaceae family, commonly found in various environments. This opportunistic pathogen can cause urinary tract infections, respiratory infections and septicaemia, but endocarditis is particularly rare and concerning due to its rapid and devastating progression. We report the first documented case worldwide of infective endocarditis (IE) caused by S. marcescens producing NDM-type carbapenemase, and the second reported case of S. marcescens endocarditis in a preterm infant. The patient was a preterm male infant born at 34 weeks of gestation, from a triplet pregnancy, admitted to the neonatal intensive care unit on day 2 of life for respiratory distress. The mother, aged 39, had undiagnosed gestational diabetes. Premature rupture of membranes had occurred 10 days before delivery, necessitating prophylactic treatment with amoxicillin. On day 4 of life, the newborn developed a fever with elevated C-reactive protein (CRP) levels and leucocytosis, leading to antibiotic therapy with colistin, imipenem and amikacin. Blood cultures revealed the presence of carbapenemase-producing S. marcescens sensitive to fluoroquinolones. A cardiac ultrasound showed a vegetation on the mitral valve, confirming the diagnosis of IE. Despite intensive treatment, the newborn died on day 16 of life due to septic shock. This rare case of endocarditis caused by S. marcescens highlights the severity of this infection in preterm infants. Treatment relies on appropriate antibiotic therapy. Prevention requires strict hygiene measures. Further research is needed to establish optimal therapeutic recommendations.
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Fortuitous discovery of a microfilaria of the genus Loa loa during a routine blood smear at the Hematology Laboratory of the Mohamed V Military Instruction Hospital in Rabat: a case report
show More to view fulltext, buy and share links for: show Less to hide fulltext, buy and share links for:Loiasis is a parasitic infection transmitted by a vector, specifically through the bites of Chrysops genus tabanid flies. It is often associated with marked and persistent eosinophilia in affected individuals. We report the case of a 28-year-old Cameroonian male patient. His medical history includes an episode of malaria treated on an outpatient basis. As part of a diving internship in Morocco, the young serviceman underwent a medical fitness examination at the Medical Expertise Center for Aircrew Personnel of the Mohamed V Military Hospital, which included a biological assessment. This revealed a mildly elevated bilirubin level, lactate dehydrogenase activity at the upper limit of normal and eosinophilia at 1500 µl–1, without anaemia or thrombocytopaenia. A blood smear was prepared and stained with May-Grünwald Giemsa, revealing the presence of several small worms, with an appearance consistent with Loa loa microfilariae. This case of L. loa, identified in the haematology laboratory, is one of the rare diagnoses in Morocco. Therefore, biologists need to remain vigilant and carry out a thorough analysis of the blood count and blood smear.
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Crystalline lens dislocation as a presenting sign of Streptococcus pyogenes invasive infections
show More to view fulltext, buy and share links for: show Less to hide fulltext, buy and share links for:Introduction. To describe two cases of crystalline lens dislocation as a presenting feature of invasive group A Streptococcus (GAS) infection and its management.
Case presentation. We report on a 58-year-old woman and a 36-year-old man who presented in 2024 with acute vision loss and severe ocular and systemic symptoms. Both patients were found to have lens dislocation and were diagnosed with invasive GAS infection. The 58-year-old woman had a complicated clinical course leading to enucleation, while the 36-year-old man responded favourably to early and aggressive treatment with systemic and intravitreal antibiotics. The responsible GAS strains were sequence type (ST) 28 and ST433, respectively.
Conclusion. These cases highlight the importance of recognizing crystalline lens dislocation as a potential sign of ocular GAS infection. Two specific strain types of GAS associated with these findings, ST28 and ST433, are reported. In patients with GAS sepsis presenting with corneal oedema and zonular loss, clinicians should immediately initiate treatment, including intravitreal antibiotic injections and systemic therapy. Prompt and aggressive management can be crucial in preserving ocular structures.
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A cautionary tale of false-positive histoplasma urine antigen in an HIV patient: a case report
show More to view fulltext, buy and share links for: show Less to hide fulltext, buy and share links for:Introduction. Coccidioidomycosis, or Valley fever, is a fungal disease caused by Coccidioides species, prevalent in parts of the southwestern United States. It usually results from inhaling spores from soil and is a common cause of pneumonia in these regions.
Case Presentation. We present a unique case of coccidioidomycosis in an immunodeficient male patient secondary to human immunodeficiency virus infection with poor adherence to anti-retroviral treatment. After presenting with non-specific symptoms and pre-syncope, he was initially diagnosed with pneumonia based on chest X-ray findings, but his symptoms failed to improve with antibiotics. He was treated for presumed pulmonary histoplasmosis following a positive histoplasma urine antigen test. However, the patient worsened clinically. Following a computed tomography scan demonstrating a large necrotic lung consolidation, fungal stain and culture of tissue biopsied through endobronchial ultrasound confirmed coccidioidomycosis. The patient received 2 weeks of liposomal amphotericin with clinical improvement before discharge with itraconazole.
Conclusion. The histoplasma antigen test can be falsely positive due to cross-reaction with other fungal infections like blastomycosis, paracoccidioidomycosis or talaromycosis, and less frequently, coccidioidomycosis or aspergillosis. Diagnosis of coccidioidomycosis requires a high index of suspicion outside the expected geographic distribution in the appropriate clinical setting. Our case highlights the risk of false-positive antigen test results and the importance of invasive diagnostics, including bronchoscopy to obtain fungal cultures, if the diagnosis remains uncertain.
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- Corrigenda
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Prevalence and resistance pattern of uropathogens from community settings of different regions: an experience from India
Sarita Mohapatra, Rajashree Panigrahy, Vibhor Tak, Shwetha J. V., Sneha K. C., Susmita Chaudhuri, Swati Pundir, Deepak Kocher, Hitender Gautam, Seema Sood, Bimal Kumar Das, Arti Kapil, Pankaj Hari, Arvind Kumar, Rajesh Kumari, Mani Kalaivani, Ambica R., Harshal Ramesh Salve, Sumit Malhotra and Shashi Kant
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Lower respiratory tract microbiome composition and community interactions in smokers
Michael Campos, Trevor Cickovski, Mitch Fernandez, Melita Jaric, Adam Wanner, Gregory Holt, Elio Donna, Eliana Mendes, Eugenia Silva-Herzog, Lisa Schneper, Jonathan Segal, David Moraga Amador, Juan Daniel Riveros, Vanessa Aguiar-Pulido, Santanu Banerjee, Matthias Salathe, Kalai Mathee and Giri Narasimhan
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