Diversity of the diploid sequence type of Candida albicans clinical isolates from a tertiary-care hospital in Mwanza, Tanzania.

Geographical strain variations of Candida albicans causing different clinical conditions in susceptible individuals have been reported. In this study, the distribution of diploid sequence type of C. albicans was investigated in Mwanza, Tanzania. A total of 64 C. albicans were selected on the basis of their antifungal susceptibility patterns, followed by multilocus sequence typing (MLST) to establish the circulating sequence types (STs). Forty-eight MLST were obtained out of 64 isolates amounting to 75% population structure differences. Out of these STs, 27 (56.3%) were new diploid ST types. C. albicans isolates with new ST were more diverse than isolates with known STs (27/29, 93.1% vs. 21/35, 60%, p 0.002). In conclusion, C. albicans from clinical specimens were highly diverse, with more than half of the detected diploid ST not previously reported in the MLST database, thus confirming the genetic differences of C. albicans from different geographical regions.

Deciphering risk factors for blood stream infections, bacteria species and antimicrobial resistance profiles among children under five years of age in North-Western Tanzania: a multicentre study in a cascade of referral health care system.

BACKGROUND: Blood stream infections (BSIs) cause a complex cascade of inflammatory events, resulting in significant morbidity and mortality in children in Tanzania. This study was designed to delineate circulating bacterial species, antimicrobial resistance (AMR) profiles and risk factors for BSIs and mortality among children in the cascade of referral health care facilities so as to guide comprehensive BSIs management. METHODS: A multiple cross sectional analytical study was conducted between July 20, 2016 to October 04, 2017 involving 950 children less than five years of age in the North-western part of Tanzania. Children with clinical features suggestive of BSIs were included. Demographic, clinical and laboratory information on culture and antimicrobial susceptibility testing was collected from children; and analyzed using STATA version 13.0 software. RESULTS: The prevalence of BSIs among children was 14.2% (95% CI: 12.1-16.6%), with specific prevalence in the district, regional and tertiary hospitals being 8.3, 6.4 and 20.0%, respectively. The most common bacterial pathogens isolated from 135 culture-positive children were Klebsiella pneumoniae (55, 40.4%), Staphylococcus aureus (23, 17.0%), and Escherichia coli (17, 12.6%). Multi-drug resistance (MDR) was higher in isolates from children at Bugando Medical Centre (BMC) tertiary hospital than isolates from district and regional hospitals [OR (95% CI): 6.36 (2.15-18.76); p = 0.001]. Independent risk factors for BSIs were neonatal period [OR (95% CI): 1.93 (1.07-3.48); p = 0.003] and admission at BMC [2.01 (1.08-3.74); p = 0.028)]. Approximately 6.6% (61/932) of children died, and risk factors for mortality were found to be children attending BMC [OR (95% CI): 4.95 (1.95-12.5); p = 0.001)], neonatal period [OR (95% CI): 2.25 (1.02-5.00); p = 0.045)], and children who had blood culture positive results [OR (95% CI): 1.95 (1.07-3.56); p = 0.028)]. CONCLUSIONS: The prevalence of BSIs (14.2%) in this multi-centre study is high and predominantly caused by the MDR K. pneumoniae. Priority interventional measures to combat BSIs and mortality, specifically among neonates at BMC are urgently recommended.

Multicentre evaluation of significant bacteriuria among pregnant women in the cascade of referral healthcare system in North-western Tanzania: Bacterial pathogens, antimicrobial resistance profiles and predictors.

OBJECTIVES: The aim of this multicentre study was to evaluate the magnitude of significant bacteriuria (SB) as well as the implicated bacterial pathogens, antimicrobial resistance (AMR) profiles and risk factors for SB among pregnant women attending different levels of healthcare facilities (HCFs) in Tanzania in order to guide antimicrobial therapy and preventive measures. METHODS: Information on sociodemographic and clinical characteristics, midstream urine culture and antimicrobial susceptibility testing was collected from 1828 pregnant women between March 2016 and May 2017. Data were analysed using STATA v.13.0 software. RESULTS: The prevalence of SB among pregnant women was 17.7% (323/1828; 95% CI 16.0-19.5%), with a predominance of Escherichia coli (164/323; 50.8%), Klebsiella spp. (55/323; 17.0%) and Staphylococcus aureus (28/323; 8.7%). Moreover, 37.5% (121/323) of bacteria were multidrug-resistant [84.3% (102/121) Gram-negative bacteria and 15.7% (19/121) in Gram-positive bacteria; P<0.001]. Third-generation cephalosporin resistance in E. coli, Klebsiella spp. and other Enterobacteriaceae was 13.4%, 21.8% and 27.5%, respectively, and was higher in strains from a tertiary hospital (OR=3.27, 95% CI 1.02-10.49; P=0.046) compared with lower HCFs. Predictors of SB among pregnant women were lack of formal occupation, current hospital admission and presence of co-morbidities. CONCLUSIONS: The prevalence of SB among pregnant women in this study was high (17.7%) and was within the same range reported 10 years ago in a single-centre baseline study. However, there is an increase in AMR in the cascade of referral healthcare system, underscoring the need for health facility level-specific antimicrobial stewardship.

High carriage rate of extended-spectrum beta-lactamase-producing Enterobacteriaceae among patients admitted for surgery in Tanzanian hospitals with a low rate of endogenous surgical site infections.

INTRODUCTION: Despite the high reported rates of surgical site infections (SSIs) caused by extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae (EPE) in low-income countries, including Tanzania, the role of EPE carriage in subsequent occurrence of SSIs is not known. This study investigated the rates of EPE carriage among surgical patients at the time of admission and discharge, and linked EPE genotype with SSIs. METHODS: EPE were confirmed among isolates from rectal and wound/pus swabs using VITEK-2. Polymerase chain reaction and sequencing were performed to detect beta-lactamase genes. Multi-locus sequence typing was used to determine the genotypes of EPE isolates. RESULTS: Among 930 patients enrolled, EPE carriage was significantly higher on discharge than admission (36.4% vs 23.7%, P<0.001). Of 272 patients who tested negative on admission, 78 (28.7%) acquired EPE during hospitalization. History of hospital stay within the previous three months was an independent predictor of EPE acquisition [hazard ratio 2, 95% confidence interval (CI) 1.04-3.98, P=0.038]. Of the 536 patients who were successfully followed-up after surgery, 78 (14.6%, 95% CI 11.6-17.5) developed SSIs. Of 57 SSIs investigated, 33 (58%) were caused by enteric Gram-negative bacteria, of which 63.6% (21/33) were EPE. Escherichia coli sequence type (ST)131 pandemic clone and Klebsiella pneumoniae ST391 predominated among wound isolates. The blaCTX-M-15 gene was detected in 37 (97.3%) of 38 ESBL isolates. Male sex was an independent predictor of SSI (odds ratio 2.92, 95% CI 1.73-4.91, P<0.001). CONCLUSION: These findings warrant implementation of strict infection control measures, antimicrobial stewardship and exploration of the transmission dynamics of EPE in surgical wards.

Preliminary insights into the occurrence of similar clones of extended-spectrum beta-lactamase-producing bacteria in humans, animals and the environment in Tanzania: A systematic review and meta-analysis between 2005 and 2016.

The emergence and spread of extended-spectrum beta-lactamase producing Enterobacteriaceae (ESBL-PE) are complex and of the public health concern across the globe. This review aimed at assessing the ESBL-PE clones circulating in humans, animals and the environment to provide evidence-based insights for combating ESBL-PE using One Health approach. Systematic search from Medline/PubMed, Google Scholar and African Journals Online was carried out and retrieved nine eligible articles (of 131) based on phenotypic and genotypic detection of ESBL-PE between 2005 and 2016 in Tanzania. Analysis was performed using STATA 11.0 software to delineate the prevalence of ESBL-PE, phenotypic resistance profiles and clones circulating in the three interfaces. The overall prevalence of ESBL-PE in the three interfaces was 22.6% (95% CI: 21.1-24.2) with the predominance of Escherichia coli (E. coli) strains (51.6%). The majority of ESBL-PE were resistant to the commonly used antimicrobials such as trimethoprim-sulfamethoxazole and tetracycline/doxycycline, 38%-55% were resistant to ciprofloxacin and all were sensitive to meropenem/imipenem. ESBL-PE infections were more associated with deaths compared to non-ESBL-PE infections. Strikingly, E. coli ST38, ST131 and ST2852 were found to intersect variably across the three interfaces. The predominant allele, blaCTX-M-15, was found mostly in the conjugative IncF plasmids connoting transmission potential. The high prevalence of ESBL-PE and shared clones across the three interfaces, including the global E. coli ST131 clone, indicates wide and inter-compartmental spread that calls for One Health genomic-driven studies to track the resistome flow.

Detection of Extended-Spectrum Beta-Lactamase-Producing Escherichia coli in Market-Ready Chickens in Zambia.

The frequent administering of antibiotics in the treatment of poultry diseases may contribute to emergence of antimicrobial-resistant strains. The objective of this study was to detect the presence of extended-spectrum ß-lactamase- (ESBL-) producing Escherichia coli in poultry in Zambia. A total of 384 poultry samples were collected and analyzed for ESBL-producing Escherichia coli. The cultured E. coli isolates were subjected to antimicrobial susceptibility tests and the polymerase chain reaction for detection of bla CTX-M, bla SHV, and bla TEM genes. Overall 20.1%, 77/384, (95% CI; 43.2-65.5%) of total samples analyzed contained ESBL-producing Escherichia coli. The antimicrobial sensitivity test revealed that 85.7% (66/77; CI: 75.7-92) of ESBL-producing E. coli isolates conferred resistance to beta-lactam and other antimicrobial agents. These results indicate that poultry is a potential reservoir for ESBL-producing Escherichia coli. The presence of ESBL-producing Escherichia coli in poultry destined for human consumption requires strengthening of the antibiotic administering policy. This is important as antibiotic administration in food animals is gaining momentum for improved animal productivity in developing countries such as Zambia.

Pulmonary TB bacteriologically confirmed by induced sputum among children at Bugando Medical Centre, Tanzania.

BACKGROUND: The diagnosis of paediatric tuberculosis (TB) remains difficult in resource-: poor settings. OBJECTIVE: To evaluate induced sputum collection and examination using microscopy, culture and Xpert(®) MTB/RIF assay for the diagnosis of pulmonary TB (PTB) in a Tanzanian hospital vs. PTB diagnosis using clinical scoring tools alone. METHODS: We conducted a cross-sectional study from October 2013 to April 2014 at our hospital in northwestern Tanzania. Children presumed to have TB were assessed using four TB score charts and sputum examination. Sputum samples were analyzed using fluorescence microscopy, solid culture and Xpert. The number of cases microbiologically confirmed was compared to the number of TB cases suspected based on TB score charts. RESULTS: A total of 192 patients were enrolled. Sputum specimens were successfully obtained in 187 (97.4%) patients without any major complications. Ten (5.2%) children were confirmed to have PTB by sputum examination. More than half (50-90%) of the confirmed cases were not detected by score charts alone. CONCLUSION: Sputum induction is both safe and feasible in a severely resource-limited hospital, and can lead to microbiological PTB diagnosis that would not be detected by clinical criteria alone.

Clostridium difficile infections among adults and children in Mwanza/Tanzania: is it an underappreciated pathogen among immunocompromised patients in sub-Saharan Africa?

Little is known regarding the epidemiology Clostridium difficile in developing countries. Fresh stool samples from patients with diarrhoea were cultured anaerobically. C. difficile was detected in nine (6.4%) of 141 (95% confidence interval 4.2-13.1), of which seven (77.8%) were from children. HIV infection, prolonged hospitalization and antibiotic use were independent factors associated with the occurrence of C. difficile in the gastrointestinal tract. Two of the toxigenic isolates were typed as ribotype 045, and the other two had unknown ribotype. All C. difficile isolates were susceptible to metronidazole, moxifloxacin and clarithromycin, while three isolates were resistant to clarithromycin. C. difficile may be an important pathogen causing diarrhoea in sub-Saharan Africa among immunocompromised patients.

Multiple ST clonal complexes, with a predominance of ST131, of Escherichia coli harbouring blaCTX-M-15 in a tertiary hospital in Tanzania.

The molecular epidemiology of 32 non-duplicate, CTX-M-15 extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli strains, isolated from clinical samples, was investigated. Multilocus sequence typing revealed multiple sequence type clonal complexes: ST131 (12), ST405 (4), ST638 (3), ST38 (2), ST827 (2), ST224 (1), ST648 (1), ST46 (1) and two new sequence type clonal complexes (1845 and 1848) in 22 pulsed field gel electrophoresis clusters. The bla(CTX-M-15) gene was located on conjugative IncF plasmids. This is the first report of the worldwide emerging clonal complex ST131 linked to bla(CTX-M-15) in Tanzania and demonstrates the need for constant surveillance in developing countries to prevent the spread of these multiresistant isolates.

Role of microscopic examination of stool specimens in the diagnosis of campylobacter infection from children with acute diarrhoea in Kampala, Uganda.

Campylobacter species are a frequent cause of enteritis and less often of extraintestinal infections in humans. The diagnosis of campylobacter infection depends mainly on culture which is difficult and expensive to be done as routine in most clinical microbiology laboratories in the developing countries. This study was conducted to determine the sensitivity and specificity of Gram-stain of the stool in diagnosis of campylobacter infection, using culture as the gold standard. A total of 226 stool specimens were obtained from children with acute diarrhoea, attending Mulago Hospital in Kampala, Uganda. Stool smears were made and conventional Gram stain done using 0.3% carbol-fuschin as counter stain for 5 minutes. Mucous part of the stool was cultured in Charcoal Ceferaperazone Deoxycholate Agar and blood contained selective media. A total of 21 stool samples (9.3%) were positive by culture and 17 (7.5%) by Gram stain. Sensitivity and specificity of Gram stain in the diagnosis of campylobacter infection was 76% and 99.5%, respectively with positive predictive value of 94.1%. A total of 127 (56.2%) had white blood cells (WBC) in stool and there was strong association between WBC in stool and the presence of campylobacter infection (P=0.001). Gram stain is a good alternative in diagnosis of campylobacter infection in place where facilities for culture are limited.

Prevalence of urinary tract infection among pregnant women at Bugando Medical Centre, Mwanza, Tanzania.

Urinary tract infections (UTI) are the most common bacterial infections during pregnancy and these infections. Untreated UTI can be associated with serious obstetric complications. This cross-sectional study was carried out to determine the prevalence of UTI among symptomatic and asymptomatic pregnant women attending Bugando Medical centre (BMC) in Mwanza, Tanzania. A total of 247 pregnant women were enrolled, of these 78 (31.5%) were symptomatic and 169 (68.4%) asymptomatic. UTI was diagnosed using mid stream urine (MSU) culture on standard culture media and urinalysis was done using rapid dip stick. The prevalence of bacteriuria among symptomatic and asymptomatic pregnant women were 17.9% and 13.0% respectively, with no significant difference between the two groups (p = 0.307). Using univariate analysis there was no association of parity (p = 0.825), gestational age (p = 0.173), education (p = 0.615), age (p = 0.211) and marital status (p = 0.949) with bacteriuria. The sensitivity and specificity of urine dipstick was 38.9% and 86.7% respectively. Escherichia coli (47.2%) and Enterococcus spp (22.2%) were the most commonly recovered pathogens. The rate of resistance of Escherichia coli to ampicillin, tetracycline, sulfamethaxazole/trimethoprim, gentamicin, ciprofloxacin, nitrofurantoin, ceftriaxone, and imipenem were 53%, 58.8%, 64.7%, 5.9%, 11.8%, 5.9%, 29.4% and 0%, respectively. In conclusion, asymptomatic bacteriuria among pregnant women is prevalent in our setting and majority of Escherichia coli are resistant to ampicillin, tetracycline, SXT and ceftriaxone. Due to low sensitivity of rapid dip stick, routine urine culture and susceptibility testing is recommended to all pregnant women at booking.

Campylobacter spp among children with acute diarrhea attending Mulago hospital in Kampala--Uganda.

BACKGROUND: Campylobacter infections occur worldwide. A recent study in Kampala, Uganda, found that 87% of broiler chickens had Campylobacter jejuni; these are potential source of human infection. Isolation rate in developing countries is between 5-35%. This study aimed at finding prevalence of children with campylobacter infection among children with acute diarrhea attending Mulago hospital. OBJECTIVE: The objective was to establish the proportion of children infected with Campylobacter spp among children with acute diarrhea at Mulago hospital. METHODS: A crossectional study from July to October 2005 was conducted involved 226 children with acute diarrhea. Serial sampling was done a total of 226 stool specimens were obtained and cultured on selective media. Identification was done using biochemical test and susceptibility using standard discs diffusion method. RESULTS: Campylobacter spp were isolated in 21 (9.3%) of 226 stool specimens analyzed. Campylobacter jejuni 17 (80.9%), Campylobacter lari 2 (9.5%), Campylobacter coli 1 (4.5%) and Campylobacter jejuni/coli 1(4.5%). All Campylobacter isolates were sensitive to erythromycin, and 20% had intermediate resistance to Ampicillin. CONCLUSION: Campylobacter spp are prevalent among children with acute diarrhea in Kampala- Uganda. A large multicenter study should be undertaken so that the extent of campylobacter infection in our setting can be established.