Molecular identification and anti-malarial drug resistance profile of Plasmodium falciparum from patients attending Kisoro Hospital, southwestern Uganda

Background The evolution of malaria infection has necessitated the development of highly sensitive diagnostic assays, as well as the use of dried blood spots (DBS) as a potential source of deoxyribonucleic acid (DNA) yield for polymerase chain reaction (PCR) assays. This study identified the different Plasmodium species in malaria-positive patients, and the anti-malarial drug resistance profile for Plasmodium falciparum using DBS samples collected from patients attending Kisoro Hospital in Kisoro district, Southwestern Uganda. Methods The blood samples were prospectively collected from patients diagnosed with malaria to make DBS, which were then used to extract DNA for real-time PCR and high-resolution melting (HRM) analysis. Plasmodium species were identified by comparing the control and test samples using HRM-PCR derivative curves. Plasmodium falciparum chloroquine (CQ) resistance transporter (pfcrt) and kelch13 to screen the samples for anti-malarial resistance markers. The HRM-PCR derivative curve was used to present a summary distribution of the different Plasmodium species as well as the anti-malarial drug profile. Results Of the 152 participants sampled, 98 (64.5%) were females. The average age of the participants was 34.9 years (range: 2 months–81 years). There were 134 samples that showed PCR amplification, confirming the species as Plasmodium. Plasmodium falciparum (N = 122), Plasmodium malariae (N = 6), Plasmodium ovale (N = 4), and Plasmodium vivax (N = 2) were the various Plasmodium species and their proportions. The results showed that 87 (71.3%) of the samples were sensitive strains/wild type (CVMNK), 4 (3.3%) were resistant haplotypes (SVMNT), and 31 (25.4%) were resistant haplotypes (CVIET). Kelch13 C580Y mutation was not detected. Conclusion The community served by Kisoro hospital has a high Plasmodium species burden, according to this study. Plasmodium falciparum was the dominant species, and it has shown that resistance to chloroquine is decreasing in the region. Based on this, molecular identification of Plasmodium species is critical for better clinical management. Besides, DBS is an appropriate medium for DNA preservation and storage for future epidemiological studies.

Classification of dairy properties according to the geometric mean of the somatic cell count and its antimicrobial resistance profile

Milk and its derivatives are highly consumed foods worldwide, with recognized nutritional importance. The search for the production of products with superior quality is constant. For the present work, 26 milk-producing properties were selected, with a total of 506 milk samples collected during the period from October 2019 to May 2020 being evaluated. The objective of this study was to evaluate the quality of milk produced in dairy properties in the region west Paraná, classified as good or bad based on the results of the Somatic Cell Count (SCC) and through sampling (n = 10) to evaluate the resistance profile of enterobacteria and Staphylococcus spp. isolated from milk samples, in addition to the presence of the mecA gene in strains of Staphylococcus spp. resistant to oxacillin. There were significant differences between the good and bad properties for the levels of lactose, SCC (cell/mL), and Standard Plate Count (SPC) (CFU/mL). The strains of Staphylococcus spp. showed differences in the percentage of resistance in relation to the good and bad properties for antibiotics: tetracycline, ciprofloxacin, oxacillin, amikacin, clindamycin, gentamycin, and erythromycin. The mecA gene was not detected in any of the coagulase-negative Staphylococcus isolates that showed resistance to oxacillin. For enterobacteria, the isolated species differed in relation to the classification of properties, with predominance for Escherichia coli (40%) for properties classified as bad and Hafnia alvei (40%) for those classified as good. The percentage of antibiotic resistance compared to enterobacteria isolates was higher in properties classified as good. Monitoring through microbial culture and antibiogram is extremely important, favoring the correct choice for the treatment of animals with a reduced selection of resistant strains.

Prevalence of Bacillus cereus in Ready-to-Eat Foods in Northern Perak, Malaysia

Food poisoning cases in Malaysia showed an increasing trend every year where 496 episodes were reported in 2018 as compared to 401 episodes in the same week of the year 2017. Bacillus cereus is one of the foodborne pathogens related to food poisoning cases in Malaysia. The main cause for the outbreak of B. cereus is the unregulated temperature during holding time. This study was conducted to detect the presence of aerobic bacteria and B. cereus present in ready-to-eat food in Northern Perak. A total of 83 food samples were collected and analyzed for the microbial count. The result shows that aerobic bacteria and B. cereus were detected in 28% of the samples. B. cereus count in food samples tested ranged from 100 cfu/g to 42000 cfu/g, whereas the aerobic bacteria recorded a range of 500 cfu/g to 2100000 cfu/g. The highest percentage of B. cereus was found in rice-based food, followed by meat, poultry, and gravy dishes. Positive colonies of B. cereus were further tested for anti-microbial resistance profile. Most B. cereus isolates showed resistance to tetracycline and clindamycin.

Antimicrobial Resistance Profile of MDR & Non-MDR Meropenem-Resistant Pseudomonas aeruginosa Isolates of Patients in Intensive Care Unit of Tertiary Hospital

Pseudomonas aeruginosa is one of the gram-negative bacteria that causes infection in the Intensive Care Unit (ICU) which is easily resistant. Patients infected with carbapenem-resistant P. aeruginosa are predicted to have a poor prognosis. This study aims to know the resistance profile of meropenem-resistant P. aeruginosa in the ICU. The results of this study can be used as a measure on the success of antimicrobial resistance control, infection control programs and become a reference for empirical therapy in the ICU. This study used a cross-sectional retrospective descriptive research method and was carried out at the Clinical Microbiology Laboratory of Sanglah Hospital Denpasar for three years, from 2018 to 2020. The results showed 38 of the 93 isolates of P. aeruginosa in the ICU were resistant to meropenem and were derived from sputum and urine. The percentage of meropenem-resistant P. aeruginosa isolates was higher in the multi-drug-resistant group and mostly came from sputum specimens. In 2018, Non-MDR meropenem-resistant P. aeruginosa isolates was that 100% sensitive to all other antibiotics used to treat P. aeruginosa infections, including; ceftazidime, cefepime, ciprofloxacin, gentamicin, amikacin, and piperacillin-tazobactam. In 2019 no meropenem-resistant P. aeruginosa isolates were found. In 2020, its sensitivity to antibiotics ceftazidime and piperacillin-tazobactam was 20.0%, ciprofloxacin 60.0% and to antibiotics gentamicin and amikacin 100%. MDR meropenem-resistant P. aeruginosa isolates in 2018 were still sensitive to ceftazidime (15.4%) and amikacin (69.2%) antibiotics, while in 2019 they were only sensitive to amikacin (37.5%). In 2020, P. aeruginosa isolates were sensitive to the antibiotics ceftazidime and cefepime (11.1%), piperacillin-tazobactam (22.2%), and amikacin (88.9%). Amikacin may be the choice of treatment for MDR meropenem-resistant P. aeruginosa.

Whole Genome Sequencing and Beta-Lactam Resistant Genes in Klebsiella pneumoniae Strains Clinically Isolated from CU Shah Hospital, Gujarat, India

A simple explanation for antimicrobial-resistant opportunistic infections in immunocompromised patients is Klebsiella pneumoniae which gradually being associated in insidious infections globally with high mortality rate. Eight hundred fifty-six antibiotic resistant K. pneumoniae isolates were collected over 3 years period (from different wards and different specimens) from the Microbiology department of C.U. Shah hospital, whose AST checked by Kirby Bauer disk diffusion method. To study AMR genes, virulome, interference of virulence gene with resistance gene, phylogenomic; 6 clinical isolates were proceeded for whole genome sequencing and bio informatics analysis. Klebsiella pneumoniae is a multidrug-resistant (MDR) opportunistic and one of delegate of ESKAPE pathogens groups. This pathogen causes nosocomial infections, urinary tract infections, liver abscesses, wound infections, meningitis. These strains obtain a multidrug resistant phenotype by way of horizontal transfer of ARG transported by either transposons or plasmids. This transfer is generally facilitated by Integrons. In this study antibiotic resistance profile and antibiotic resistance genes analysis as well as virulence gene of K. pneumoniae strains were investigated. The study was carried out using 853 clinical isolates collected during 3 years from C.U. Shah hospital of Surendranagar. Antibiotic resistance profile test was carried out by the VITEK 2 against 21 antibiotics. Out of that 6 samples were proceed for DNA extraction, WGS illumina sequencer and analysis of those raw sequences by TORMES pipeline. In this study antibiotic resistance profile included 13 beta lactam antibiotics which classified under 3 class (Penicillin, Cephalosporin, Carbapenem) of beta lactam and in AMR gene study got total 15 different ESBL resistance genes from 6 different klebsiella pneumoniae strain. All these genes detected with more than 90% identity by CARD. (TORMES Pipeline) CTX-M-15, NDM-5, OKP-B-6, PDC-2, OXA-1, OXA-181, OXA-362, OXA-50, OXA-9, SHV-1, SHV-11, SHV-187, TEM-1, TEM-150. In this study, we’ve analyzed the pattern of antibiotic resistance pattern as a phenotypic characteristic and antibiotic resistance genes as genotypic characteristic and co related the results. As multidrug resistance is a worrying matter, constant observation and regular clinical recognition of resistant bacteria are essential to avoid terrible public health incidents. So, our data should be inferred as a warning for need for prevention and control of the MDR K. pneumoniae in hospital settings.

Prevalence and antibiotic resistance profile of Staphylococcus aureus and Eschericia coli among patients attending urology clinic of Dalhatu Araf specialist hospital (DASH) Lafia, Nasarawa State, Nigeria

No abstract.