Safety, tolerability, and immunogenicity of an oral inactivated ETEC vaccine (ETVAX®) with dmLT adjuvant in healthy adults and children in Zambia: An age descending randomised, placebo-controlled trial.

BACKGROUND: Enterotoxigenic Escherichia coli (ETEC) is an important cause of moderate to severe diarrhoea in children for which there is no licensed vaccine. We evaluated ETVAX®, an oral, inactivated ETEC vaccine containing four E. coli strains over-expressing the major colonization factors CFA/I, CS3, CS5, and CS6, a toxoid (LCTBA) and double mutant heat-labile enterotoxin (dmLT) adjuvant for safety, tolerability, and immunogenicity. METHODS: A double-blind, placebo-controlled, age-descending, dose-finding trial was undertaken in 40 adults, 60 children aged 10-23 months, and 146 aged 6-9 months. Adults received one full dose of ETVAX® and children received 3 doses of either 1/4 or 1/8 dose. Safety was evaluated as solicited and unsolicited events for 7 days following vaccination. Immunogenicity was assessed by evaluation of plasma IgA antibody responses to CFA/I, CS3, CS5, CS6, and LTB, and IgG responses to LTB. RESULTS: Solicited adverse events were mostly mild or moderate with only 2 severe fever reports which were unrelated to the vaccine. The most common events were abdominal pain in adults (26.7 % in vaccinees vs 20 % in placebos), and fever in children aged 6-9 months (44 % vs 54  %). Dosage, number of vaccinations and decreasing age had no influence on severity or frequency of adverse events. The vaccine induced plasma IgA and IgG responses against LTB in 100 % of the adults and 80-90 % of the children. In the 6-23 months cohort, IgA responses to more than 3 vaccine antigens after 3 doses determined as ≥2-fold rise was significantly higher for 1/4 dose compared to placebo (56.7 % vs 27.2 %, p = 0.01). In the 6-9 months cohort, responses to the 1/4 dose were significantly higher than 1/8 dose after 3 rather than 2 doses. CONCLUSION: ETVAX® was safe, tolerable, and immunogenic in Zambian adults and children. The 1/4 dose induced significantly stronger IgA responses and is recommended for evaluation of protection in children. CLINICAL TRIALS REGISTRATION: The trial is registered with the Pan African Clinical Trials Registry (PACTR Ref. 201905764389804) and a description of this clinical trial is available on: https://pactr.samrc.ac.za/Trial Design.

Bartonella diversity and zoonotic potential in indigenous Tete Veld rats (Aethomys ineptus) from South Africa.

Bartonellosis is a vector-borne disease that is often misdiagnosed due to a broad range of clinical symptoms, compounded by a lack of awareness regarding the prevalence, diversity and public health impacts of regional strains. Despite recent PCR-based confirmation of Bartonella in 9.7% of non-malarial, acute febrile patients in South Africa, data regarding reservoirs of infection are limited. As the majority of Bartonella species described to date are associated with rodent species globally, including zoonotic species such as B. elizabethae, and as rodent biodiversity is high in southern Africa, we evaluated Bartonella in the Tete Veld rat (Aethomys ineptus), a highly adaptable murid rodent that thrives in both natural and commensal settings. These rodents are infested with a broad range of ectoparasite species, and often occur in sympatry with Micaelamys namaquensis, an indigenous rodent previously shown to host B. elizabethae. DNA extracts from heart samples of 75 A. ineptus trapped over an eight-month period, from the Roodeplaat Nature Reserve (RNR), were evaluated using a multi-locus sequence analysis (MLSA) approach. Nucleotide sequencing and phylogenetic analyses of individual (gltA, ribC, rpoB and nuoG) and concatenated gene datasets confirmed the presence of three discrete Bartonella lineages (I-III). Lineages I and II, are genetically distinct from all currently recognised Bartonella species but cluster with strains present in other indigenous rodents from South and East Africa, whereas lineage III contained B. elizabethae, a zoonotic species associated with Rattus species globally. Records confirming R. tanezumi presence in this nature reserve, which is situated in close proximity to Pretoria, the administrative capital of South Africa, suggests the likelihood of spill-over from invasive to indigenous species. These results together with the high levels of infection (86.7%) and co-infection (33.8%), indicate that A. ineptus is a natural reservoir for multiple Bartonella species in South Africa, including one with zoonotic potential.

Molecular assessment of Bartonella in Gerbillus nanus from Saudi Arabia reveals high levels of prevalence, diversity and co-infection.

Bartonellae bacteria are associated with several re-emerging human diseases. These vector-borne pathogens have a global distribution, yet data on Bartonella prevalence and diversity in the Arabian Peninsula are limited. In this study we assessed the Bartonella infection status of the Baluchistan gerbil (Gerbillus nanus), a species associated with pastoral communities throughout the Middle East region, using a multi-gene PCR screening approach. The results demonstrated that 94 (68.1%) of the 138 gerbils trapped on a monthly basis, over a period of one year, were PCR-positive. Sequencing of the gltA gene region confirmed the presence of four discrete Bartonella lineages (I-IV) and high levels of co-infection (33.0%). Each of the four lineages, varied in overall abundance (7.5%-47.9%) and had discernible seasonal peaks. Bartonella status was significantly correlated with ectoparasite presence, but not with sex, nor with season. Statistical analyses further revealed that co-infected individuals had a significantly higher relative body condition. Multi-locus sequence analysis (MLSA) performed with a concatenated dataset of three genetic loci (gltA, nuoG, and rpoB), 1452 nucleotides (nt) in length confirmed that lineage IV, which occurred in 24 PCR-positive animals (25.5%), is most closely related to zoonotic B. elizabethae. The remaining three lineages (I-III) formed a monophyletic clade which, on the basis of gltA was shown to contain bartonellae from diverse Gerbillinae species from the Middle East, suggestive of a gerbil-associated species complex in this region. Lineage I was identical to a Candidatus B. sanaae strain identified previously in Bushy-tailed jirds (Sekeetamys calurus) from Egypt, wherease MLSA indicate that lineages II and III are novel. The high levels of infection and co-infection, together with the presence of multiple Bartonella lineages indicate that Gerbillus nanus is likely a natural reservoir of Bartonella in the Arabian Peninsula.