Background Antimicrobial resistance (AMR) of Neisseria gonorrhoeae is a threat to public health as strains have developed resistance to antimicrobials available for the treatment of gonorrhea. Whole genome sequencing (WGS) can detect and predict antimicrobial resistance to enhance the control and prevention of gonorrhea. Data on the molecular epidemiology of N. gonorrhoeae is sparse in Zambia. This study aimed to determine the genetic diversity of N. gonorrhoeae isolated from patients attending sexually transmitted infection (STI) clinics in Lusaka, Zambia. Methods A cross-sectional study that sequenced 38 N. gonorrhoeae isolated from 122 patients with gonorrhea from 2019 to 2020 was conducted. The AMR profiles were determined by the E-test, and the DNA was extracted using the NucliSens easyMaG magnetic device. Whole genome sequencing was performed on the Illumina NextSeq550 platform. The Bacterial analysis pipeline (BAP) that is readily available at: https://cge.cbs.dtu.dk/services/CGEpipeline-1.1 was used for the identification of the species, assembling the genome, multi-locus sequence typing (MLST), detection of plasmids and AMR genes. Phylogeny by single nucleotide polymorphisms (SNPs) was determined with the CCphylo dataset. Results The most frequent STs with 18.4% of isolates each were ST 7363 , ST 1921 and ST 1582 , followed by ST 1583 (13%), novel ST 17026 (7.9%), ST 1588 (7.9%), ST 1596 (5.3%), ST 11181 (5.3%), ST 11750 (2.6/%) and ST 11241 (2.6%) among the 38 genotyped isolates. The blaTeM-1B and tetM (55%) was the most prevalent combination of AMR genes, followed by blaTeM-1B (18.4%), tetM (15.8%), and the combination of blaTeM-1B , ermT , and tetL was 2.6% of the isolates. The AMR phenotypes were predicted in ciprofloxacin, penicillin, tetracycline, azithromycin, and cefixime. The combination of mutations 23.7% was gryA (S91F), parC (E91G), ponA (L421) and rpsJ (V57M), followed by 18.4% in gyrA (S91F), ponA (L421P), rpsJ (V57M), and 18.4% in gyrA (D95G, S91F), ponA (L421P), and rpsJ (V57M). The combinations in gyrA (D95G, S91F) and rpsJ (V57M), and gyrA (D95G, S91F), parC (E91F), ponA (L421P) and rpsJ (V57M) were 13.2% each of the isolates. Plasmid TEM-1 (84.2%), tetM ( 15.8%), and gonococcal genetic island (GGI) was detected in all isolates. Conclusion This study revealed remarkable heterogeneity of N. gonorrhoeae with bla TEM−1 , tetM, ponA, gyrA , and parC genes associated with high resistance to penicillin, tetracycline, and ciprofloxacin demanding revision of the standard treatment guidelines and improved antimicrobial stewardship in Zambia.

Abstract Background Neisseria gonorrhoeae, the causative agent for sexually transmitted infection (STI) gonorrhoea, has emerged with a significant public health impact on acquiring resistance to antimicrobials available for treatment. The resistance of N. gonorrhoeae limit treatment options and contributed to high morbidity associated with gonorrhoea. Data on antimicrobial resistance (AMR) profiles in N. gonorrhoeae is scares in Zambia. This study aimed to determine the antibiotic susceptibilities in N. gonorrhoeae isolates from Lusaka, Zambia. Methods A prospective cross-sectional study was conducted on 630 STI patients who presented with urethral or vaginal discharge from 2019 to 2020. Urethral and endocervical secretions were cultured on Modified Thayer Martin agar and incubated at 36 °C ± 1 °C in 5% CO2 for 24 h. Identification of N. gonorrhoeae isolates was achieved by Gram stain, oxidase, nitrocefin disk, BactiCard Neisseria, and Viteck® Compact. The AMR profiles were determined using E-test. Statistical significant was determined by Pearson’s Chi-square test, Mann-Whitney U test, or logistic regression with p-values of < 0.05 indicating significance. Results A total of 630 patients were recruited of which 46% (290/630) with the median of 29 years and interquartile range (IQR) of 19–39 years were male. The median of the females was 26 years and IQR of 15–37 years. Neisseria gonorrhoeae was isolated from 19.4% (122/630) patients of which 72.9% (89/122) were male, with highest prevalence of isolation in the age category of 25–34 years. The prevalence of resistance was high to penicillin (85.2%), tetracycline (68.9%) and ciprofloxacin (59.8%) with MIC90 of 32 µg/mL, 8 µg/mL, and 8 µg/mL respectively. The isolates had reduced susceptibility to cefixime (1.6%), spectinomycin (4.9%) and (4.9%) for azithromycin. All isolates were susceptible to ceftriaxone. Risk factors associated with AMR were douching in females (AOR 6.69, 95% CI; 1.11–40.31, p = 0.039), female gender (AOR 7.64, 95% CI; 1.11–52.33, p = 0.048), HIV-positivity (AOR 26.59, 95% CI; 3.67–192.7, p = 0.005), no condom use or unprotected sex (AOR 5.48, 95% CI; 1.17–22.75 p = 0.026), sex trading (AOR 4.19, 95% CI; 1.55–11.33, p = 0.010), and over-counter treatment of ciprofloxacin (AOR 3.44, 95% CI; 1.17–22.75, p = 0.023). Conclusion The N. gonorrhoeae resistance to penicillin, tetracycline and ciprofloxacin was high necessitating revision of the treatment guidelines. However, no resistance to ceftriaxone was detected. Therefore, monitoring of antibiotic resistance remains critical in Zambia.

Human intestinal parasitic infections are still endemic in rural and urban children in developing countries such as Zambia.We conducted a cross-sectional study to compare the occurrence of intestinal parasites in children less than 16 years between a rural hospital, Chilonga Mission Hospital CMH) in Mpika, and an urban hospital, Arthur Davison Hospital ADH) in Ndola, Zambia, in 2017.Hospital laboratory records of stool examinations results performed at both hospitals from January to December 2017 were analysed.The prevalence of intestinal parasites at CMH was 5.7% while that for ADH was 0.4%.The difference between the two prevalences was statistically significant (P<0.005).One case of Balantidium coli, not previously known to be endemic in Africa, was detected at CMH.We made recommendations on how to improve laboratory detection and control of intestinal parasites in children in endemic countries such as Zambia.