Bloodstream infections initiated by ESBL-producing Escherichia coli in neonates and infants at two hospitals in Yaoundé, Cameroon

Early diagnosis and probabilistic antibiotic therapy based on known bacterial ecology and antibiotic sensibility can reduce mortality and morbidity in pathologies caused by a bacterial infection. This study aimed at determining the prevalence and risk factors of extended-spectrum β-lactamases (ESBLs)-producing Escherichia coli isolated from blood cultures of neonates and infants population. We conducted a cross-sectional study during which pathogenic bloodstream isolates were identified. Antibiotic susceptibility test was performed on Escherichia coli isolates and phenotypic confirmation of ESBL production by Escherichia coli was performed by a double-disc synergy test. Over the course of this study, 298 blood cultures were performed and 129 (43.3%) positive cultures were obtained. Of the 129 bacterial isolates, 90 (69.7%) were Escherichia coli and 39 (30.2%) were other bacteria strains that included Klebsiella oxytoca, Streptococcus pneumonia, and Coagulase-negative staphylococci. Antibiotic susceptibility test indicated that Escherichia coli isolates were resistant to cephalosporin, penicillin, sulfonamide, and aminoglycoside antibiotic families. Further analysis indicated that 31 (34.4%) Escherichia coli strains were ESBL producers and risk factors for bloodstream infection by ESBL-producing Escherichia coli were prior to exposure to antibiotics and immune system depression. These findings clearly extend our understanding of the type of resistant initiated by ESBL-producing Escherichia coli in bloodstream infection of neonates, and infants and also provides useful information that can guide the establishment of an efficient therapeutic strategy for the community- and hospital-acquired bloodstream infection

ANTIMICROBIAL AND ANTIDIARRHEAL EFFECTS OF FOUR CAMEROON MEDICINAL PLANTS: DICHROCEPHALA INTEGRIFOLIA, DIOSCOREA PREUSII, MELENIS MINUTIFLORA, AND TRICALYSIA OKELENSIS

Objective: In order to verify the antidiarrheal activities of Dichrocephala integrifolia, Dioscorea preusii, Melenis minutiflora, Tricalysia okelensis, the in vitro antimicrobial effect on Salmonella typhi, Escherichia coli, Shigella dysenteriae A1 and Candida albicans, and the in vivo antidiarrheal activities on the intestine transit of the hydroethanol (v/v) plants extracts were studied. Methods: The antimicrobial effect of the extracts was assayed in vitro by the disc diffusion and the agar dilution methods. For in vivo study, male and female mice received per os castor oil and one hour later different doses of the extracts. Results: In vitro, D. integrifolia, D. preusii, M. minutiflora, and T. okelensis extract showed concentration-dependent activity against all the tested microbial strains with the inhibition zone ranged from 08 to 24 mm. D. integrifolia 0.5 mg/mL showed the lowest MIC on Candida albicans. The M. minutiflora and D. integrifolia MIC was 3 mg/mL on Escherichia coli and Shigella dysenteriae A1. In vivo, D. integrifolia, D. preusii, T. okelensis extract at 50 and 100 mg/kg bw and M. minutiflora 75 and 150 mg/kg bw significantly (P< 0.01) inhibited the intestinal charcoal transit. D. integrifolia 100 mg/kg bw exhibited the highest inhibition rate, 70%. Conclusion: These results suggest that D. integrifolia, D. preusii, M. minutiflora and T. okelensis extracts possesses antimicrobial and antidiarrheal properties, could be effective for diarrhea treating, and could thus justify their use in traditional medicine to treat diarrhea. D. integrifolia could have the most efficiency antimicrobial properties

Epidemiology of rotavirus diarrhea in children under 5 years in Northern Cameroon

Background: Rotavirus still remains the major cause of diarrhea in children below 5 years. No data on rotavirus epidemiology is available in the Northern regions of Cameroon. We aimed to determine the prevalence of group A rotavirus (RVA) in children below 5 years with diarrhea in two regions of Northern Cameroon (North West and Far North Regions) so as to improve our knowledge on the burden of rotavirus disease for imminent introduction of a rotavirus vaccine. Methods: Stool samples were collected during 2010 and 2011 from 390 children below 5 years presenting with diarrhea in four hospitals in Northern Cameroon and were screened for rotavirus group A by reverse transcription-polymerase chain reaction. Results: This study revealed that 42.8% of the children below 5 years had group A rotavirus infection, 46.5% in the Far North region while the North West had a prevalence of 33.9%. Of the 252 hospitalized and the 138 outpatient children, 124(49.2%) and 43(31.2%) (P=0.00085), respectively, were positive for group A rotavirus. Children below 24 months were most affected (44.7%), while the age group 49-60 months had the lowest prevalence (25%). The RVA prevalence was 44.6% in the urban and 28.9% in the rural settings of our study. It was observed that the proportion of children with diarrhea who had rotavirus accompanied with fever and vomiting in the outpatient group and inpatient group were 13.0% and 28.6% respectively, P=0.03. Conclusion: This study showed high incidence of rotavirus infection especially among hospitalized children in Northern Cameroon, suggesting that rotavirus is a major cause of childhood morbidity and mortality in this area

Streptococcus agalactiae prevalence and antimicrobial susceptibility pattern in vaginal and anorectal swabs of pregnant women at a tertiary hospital in Cameroon

Abstract Objective Group B Streptococcus (GBS) or Streptococcus agalactiae is part of the normal flora of the gut and genital tract, thus carrier pregnant women can transmit this germ to newborns which could cause early neonatal infection. In Cameroon, few studies have been conducted on GBS, thus this study sought to detect the rectal and vaginal colonization rates and the antibiotic susceptibility profile of the identified strains in pregnant women. We therefore conducted a cross-sectional study over a 6 months period analysing vaginal and anorectal samples obtained from 100 pregnant women. Cultures for the isolation of GBS were carried out according to standard microbiological methods and grouping done using the Pastorex strep Kit. All strains isolated were used for susceptibility test to various antibiotics as recommended by the French microbiology society, using the disk-diffusion method. Results The detected colonization rate was 14%. No resistance to ampicillin, oxacillin, amoxycillin–clavulanate, cefotaxime, pristinamycin, vancomycin and clindamycin was found. Just 12, 94 and 82% of strains showed sensitivity to gentamycin, erythromycin and cefoxitin respectively. This study therefore revealed that at least one out of every ten women is GBS colonized and strains showed uniform sensitivity to beta lactamines. However, decreased sensitivity to erythromycin was detected

Occurrence of blaTEM and blaCTXM Genes and Biofilm-Forming Ability among Clinical Isolates of Pseudomonas aeruginosa and Acinetobacter baumannii in Yaoundé, Cameroon

Background: Pseudomonas aeruginosa (PSA) and Acinetobacter baumannii (ACB) are non-fermentative bacteria mostly associated with nosocomial infections in humans. Objective: This study aimed to determine the antimicrobial resistance profiles and virulence gene of PSA and ACB previously isolated from humans in selected health facilities in Yaoundé, Cameroon. Methods: A total of 77 and 27 presumptive PSA and ACB isolates, respectively, were collected from the Yaoundé teaching hospital. These isolates were previously isolated from various samples including pus, blood and broncho-alveolar lavage. The identities of the isolates were determined through polymerase chain reaction (PCR) amplification of PSA and ACB specific sequences. Antimicrobial susceptibility testing (AST) was performed using the Kirby–Bauer disc diffusion method. Phenotypical expression of AmpC β-lactamases (AmpC), extended spectrum β-lactamases (ESBLs) and metallo β-Lactamases (MBLs) were determined using the combined disc method. Bacterial genomes were screened for the presence of β-lactamases blaTEM and blaCTXM genes using specific PCR. The pathogenicity of PSA and ACB was assessed through amplification of the lasB, exoA, pslA and exoS as well as OmpA and csuE virulence genes, respectively. Results: Of the 77 presumptive PSA isolates, a large proportion (75 to 97.4%) were positively identified. All (100%) of the presumptive 27 ACB harbored the ACB-specific ITS gene fragment by PCR. Twenty five percent of the PSA isolates produced ESBLs phenotypically while more than 90% of these isolates were positive for the lasB, exoA, pslA and exoS genes. A large proportion (88%) of the ACB isolates harboured the OmpA and csuE genes. blaTEM and blaCTXM were detected in 17 and 4% of PSA, respectively, while a much higher proportion (70 and 29%) of the ACB isolates possessed these resistance determinants respectively. Conclusion: Our findings reveal the occurrence of both virulence and drug-resistant determinants in clinical PSA and ACB isolates from patients in health care settings in Yaoundé, Cameroon, thus suggesting their role in the pathological conditions in patients

Prevalence of Escherichia coli Producing Extended Spectrum Beta-Lactamase (ESBL) Driven Septicaemia in Children Aged 0–2 Years in Two Districts Hospitals in Yaounde, Cameroon

Septicaemia is public health problem worldwide with a high rate of mortality among children. Epidemiological data on this phenomenon in Cameroon are still scarce. This study aimed to determine the prevalence and associated factors to septicaemia due to E. coli strains producing extended spectrum beta-lactamase (ESBL) in two hospitals in Yaoundé, Cameroon. A prospective, cross-sectional study was conducted on infants aged 0 to 2 years old at the consultation and neonatology care unit of two district hospitals of Yaoundé (UTHY and YGOPH) during a period of seven months (from August 2019 to March 2020). Each blood sample collected per infant was cultured in hemoline performance vials, and bacterial strains were identified using the Api-20 E system. In addition, an antibiotic resistant profile of isolates as well as the ESBL production were performed in accordance with the recommendations of the Antibiogram committee of the French Society of Microbiology 2019. Data were analysed in Epi-Info7.0 and for p less than 0.05, the difference was statistically significant. Of the 300 children enrolled, 130 (43.33%) were blood culture positive, and E. coli. was the most prevalent (69.23% (90/130)). Then antibiotic susceptibility test revealed that 77 over 90 E. coli strains were resistant to penicillin (with 85.55% to amoxicillin), and 34.44% were producing ESBL. Factors such as immunodeficiency, being on antibiotics, and particularly taking β-lactam were significantly associated with E. coli ESBL production ([aOR = 19.93; p = 0.0001], [aOR = 1.97; p = 0.04], and [aOR = 3.54; p = 0.01], respectively). Moreover, co-resistance to aminoglycosides, quinolones, fluoroquinolones, and cotrimoxazole were also found. This study highlighted a high prevalence of E. coli ESBL in blood samples of children aged 0–2 years in Yaoundé and prompts the development of more efficient strategies against E. coli ESBL associated mortality in infants in Cameroon

High prevalence of Panton-Valentine leukocidin positive, multidrug resistant, Methicillin-resistant Staphylococcus aureus strains circulating among clinical setups in Adamawa and Far North regions of Cameroon.

Staphylococcus aureus (S. aureus) is one of the earliest pathogens involved in human infections, responsible for a large variety of pathologies. Methicillin was the first antibiotic used to treat infections due to S. aureus but infections due to Methicillin resistant Staphylococcus aureus (MRSA) originated from hospital settings. Later, severe infections due to MRSA without any contact with the hospital environment or health care workers arose. Prevalence of MRSA has shown an alarming increase worldover including Cameroon. This Cross-sectional study was designed to evaluate the occurrence of MRSA infections in five different, most frequented Hospitals in northern Cameroon. Socio demographic data was recorded through questionnaire and different clinical specimens were collected for bacterial isolation. Identification of S. aureus was confirmed via 16s rRNA amplification using S. aureus specific primers. Molecular characterisation was performed through mecA gene, Luk PV gene screening and SCCmec typing. A total of 380 S. aureus clinical isolates were obtained of which 202 (53.2%) were nonduplicate multidrug resistant isolates containing, 45.5% MRSA. Higher number of MRSA was isolated from pus (30.4%) followed by blood culture (18.5%), and urine (17.4%). Patients aged 15 to 30 years presented high prevalence of MRSA (30.4%). Majority isolates (97.8%) carried the mecA gene, PVL toxin screening indicated 53.3% isolates carried the lukPV gene. Based on PVL detection and clinical history, CA-MRSA represented 53.3% of isolates. SCCmec typing showed that the Type IV was most prevalent (29.3%), followed by type I (23.9%). Amongst MRSA isolates high resistance to penicillin (91.1%), cotrimoxazole (86.7%), tetracycline (72.2%), and ofloxacin (70.0%) was detected. Meanwhile, rifampicin, fusidic acid, lincomycin and minocycline presented high efficacy in bacterial control. This study revealed a high prevalence of MRSA among infections due to S. aureus in Northern Cameroon. All MRSA recorded were multidrug resistant and the prevalence of CA MRSA are subsequently increasing, among population

Activity of aqueous ethanol extract of Euphorbia prostrata ait on Shigella dysenteriae type 1-induced diarrhea in rats

Aim: Euphorbia prostrata   (Euphorbiaceae) is traditionally used in Cameroon for the treatment of many diseases, including diarrhea. We investigated the acute toxicity and effect of the aqueous ethanol extract of the plant on gastrointestinal propulsion, in vitro bacterial growth and in vivo bacillary dysentery. Materials and Methods: Diarrhea was induced by oral administration of 12 x 10 8 Shigella dysenteriae type 1 (Sd1) cells. Diarrheic rats were treated for 5 days with 10, 20 or 40 mg/kg extract or 20 mg/kg norfloxacin. The faeces frequencies and the number of Sd1 were assessed and the death rate recorded. Results: The aqueous ethanol extract of E. prostrata was not toxic. In vitro, the minimal inhibitory and minimal bactericidal concentrations of the extract were 3,500 and 12,000 µg/ml, respectively. In vivo, diarrhea went along with increase in faeces frequency (P < 0.01 by the 3 rd day), increase in the bacterial population to a maximum on the 2 nd day after infection (P < 0.01). The death rate in diarrheic control group was 100% by day 6. E. prostrata extracts (20 and 40 mg/kg), like norfloxacin, reduced the bacterial growth (P < 0.01), so that by the 6 th day Sd1 density was < 100 and no death was recorded. There was a significant (P < 0.01) reduction in faeces frequencies. The extract exhibited notable (P < 0.01) inhibition of intestinal propulsion. Conclusion: The results suggest that E. prostrata possesses bactericidal and antidiarrheic properties and could be a therapeutic alternative for diarrheas of bacterial etiology

Whole-genome sequence of multi-drug resistant Pseudomonas aeruginosa strains UY1PSABAL and UY1PSABAL2 isolated from human broncho-alveolar lavage, Yaoundé, Cameroon.

Pseudomonas aeruginosa has been implicated in a wide range of post-operation wound and lung infections. A wide range of acquired resistance and virulence markers indicate surviving strategy of P. aeruginosa. Complete-genome analysis has been identified as efficient approach towards understanding the pathogenicity of this organism. This study was designed to sequence the entire genome of P. aeruginosa UY1PSABAL and UY1PSABAL2; determine drug-resistance profiles and virulence factors of the isolates; assess factors that contribute toward stability of the genomes; and thereafter determine evolutionary relationships between the strains and other isolates from similar sources. The genomes of the MDR P. aeruginosa UY1PSABAL and UY1PSABAL2 were sequenced on the Illumina Miseq platform. The raw sequenced reads were assessed for quality using FastQC v.0.11.5 and filtered for low quality reads and adapter regions using Trimmomatic v.0.36. The de novo genome assembly was made with SPAdes v.3.13 and annotated using Prokka v.2.1.1 annotation pipeline; Rapid Annotation using Subsytems Technology (RAST) server v.2.0; and PATRIC annotation tool v.3.6.2. Antimicrobial resistance genes and virulence determinants were searched through the functional annotation data generated from Prokka, RAST and PATRIC annotation pipelines; In addition to ResFinder and Comprehensive Antibiotic Resistance Database (CARD) which were employed to determine resistance genes. The PHAge Search Tool Enhanced Release (PHASTER) web server was used for the rapid identification and annotation of prophage sequences within bacterial genome. Predictive secondary metabolites were identified with AntiSMASH v.5.0. Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and cas genes regions were also investigated with the CRISPRone and CRISPRFinder server. The genome sizes of 7.0 and 6.4 Mb were determined for UY1PSABAL and UY1PSABAL2 strains with G+C contents of 66.1% and 66.48% respectively. β-lactamines resistance genes blaPAO, aminoglycoside phosphorylating enzymes genes aph(3')-IIb, fosfomycine resistance gene fosA, vancomycin vanW and tetracycline tetA were among identified resistance genes harboured in both isolates. UY1PSABAL bore additional aph(6)-Id, aph(3'')-Ib, ciprofloxacin-modifying enzyme crpP and ribosomal methylation enzyme rmtB. Both isolates were found harbouring virulence markers such as flagella and type IV pili; and also present various type III secretion systems such as exoA, exoS, exoU, exoT. Secondary metabolites such as pyochelin and pyoverdine with iron uptake activity were found within the genomes as well as quorum-sensing systems, and various fragments for prophages and insertion sequences. Only the UY1PSABAL2 contains CRISPR-Cas system. The phylogeny revealed a very close evolutionary relationship between UY1PSABAL and the similar strain isolated from Malaysia; the same trend was observed between UY1PSABAL2 and the strain from Chinese origin. Complete analyses of the entire genomes provide a wide range of information towards understanding pathogenicity of the pathogens in question