Identification of termites from Gabon using MALDI-TOF MS.

Termites are one of the most common pests that damage wood and other cellulosic materials. Although Africa has more varieties of termite species than any other continent, few entomological studies have been conducted in Gabon. Identifying termites poses significant difficulties for entomologists. The aim of this study was to evaluate the reliability and confirm the significance of MALDI-TOF MS in identifying fresh termites collected in equatorial Africa. A total of 108 termites were collected from 13 termite nests during a field mission in 2021 in Lekedi and Bongoville, Gabon. Termites were morphologically identified and subjected to MALDI-TOF MS, then molecular analyses using the COI and 12S rRNA genes. Four termite species were morphologically identified in this study: Pseudacanthotermes militaris, Macrotermes muelleri, Macrotermes nobilis, and Noditermes indoensis. However, when using molecular biology, only three species were identified, namely Macrotermes bellicosus, P. militaris, and N. indoensis, because the specimens initially identified as M. muelleri and M. nobilis were found to be M. bellicosus. The MALDI-TOF MS spectral profiles of the termites were all of good quality, with intra-species reproducibility and inter-species specificity. The spectra of 98 termites were blind tested against our upgraded database, which included the spectra of ten termite specimens. All tested spectra were correctly matched to their respective species, with log score values (LSVs) ranging from 1.649 to 2.592. The mean LSV was 2.215 ± 0.203, and the median was 2.241. However, 95.91% (94/98) of our spectra had LSVs above 1.8. This study demonstrates how a proteomic approach can overcome termites' molecular and morphological identification limitations and serve as a useful taxonomic tool.

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry traces the geographical source of Biomphalaria pfeifferi and Bulinus forskalii, involved in schistosomiasis transmission.

BACKGROUND: Freshwater snails of the genera Bulinus spp., Biomphalaria spp., and Oncomelania spp. are the main intermediate hosts of human and animal schistosomiasis. Identification of these snails has long been based on morphological and/or genomic criteria, which have their limitations. These limitations include a lack of precision for the morphological tool and cost and time for the DNA-based approach. Recently, Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight (MALDI-TOF) mass spectrometry, a new tool used which is routinely in clinical microbiology, has emerged in the field of malacology for the identification of freshwater snails. This study aimed to evaluate the ability of MALDI-TOF MS to identify Biomphalaria pfeifferi and Bulinus forskalii snail populations according to their geographical origin. METHODS: This study was conducted on 101 Bi. pfeifferi and 81 Bu. forskalii snails collected in three distinct geographical areas of Senegal (the North-East, South-East and central part of the country), and supplemented with wild and laboratory strains. Specimens which had previously been morphologically described were identified by MALDI-TOF MS [identification log score values (LSV) ≥ 1.7], after an initial blind test using the pre-existing database. After DNA-based identification, new reference spectra of Bi. pfeifferi (n = 10) and Bu. forskalii (n = 5) from the geographical areas were added to the MALDI-TOF spectral database. The final blind test against this updated database was performed to assess identification at the geographic source level. RESULTS: MALDI-TOF MS correctly identified 92.1% of 101 Bi. pfeifferi snails and 98.8% of 81 Bu. forskalii snails. At the final blind test, 88% of 166 specimens were correctly identified according to both their species and sampling site, with LSVs ranging from 1.74 to 2.70. The geographical source was adequately identified in 90.1% of 91 Bi. pfeifferi and 85.3% of 75 Bu. forskalii samples. CONCLUSIONS: Our findings demonstrate that MALDI-TOF MS can identify and differentiate snail populations according to geographical origin. It outperforms the current DNA-based approaches in discriminating laboratory from wild strains. This inexpensive high-throughput approach is likely to further revolutionise epidemiological studies in areas which are endemic for schistosomiasis.

Description of Streptococcus thalassemiae sp. nov., a Bacterium Isolated from Human Blood.

Blood is a precious biological liquid that is normally sterile. Therefore, bacteria in the bloodstream are shown a priori anomaly. A blood culture is systematically performed to diagnose the cause of the bacteremia. Indeed, a patient received in our service had a thalassemia major and underwent a genoidentical transplant. Then, a blood test was performed to diagnose a four-day fever. In this context, we have isolated strain Marseille-Q2617 from the blood sample. It revealed a new bacterial strain that belongs to the genus Streptococcus. It is a Gram-positive coccus, nonmotile, and nonspore forming. The major fatty acid found is hexadecanoic acid, with 49.5%. A taxonomic method was used to characterize the strain by studying their phenotypic, phylogenetic, and genomic characteristics. In addition, sequence analysis of the 16S rRNA gene shows that the strain Marseille-Q2617 has 99.94% sequence similarity to Streptococcus mitis. Average nucleotide identity (ANI) analysis for strain Marseille-Q2617T showed the highest similarity of 92.9% with S. mitis. The DNA-DNA hybridization value obtained (50.2%) between strain Marseille-Q2607 and S. mitis, its closest related species, was below the recommended threshold (<70%). Strain Marseille-Q2617T has a genome size of 2.02 Mbp with 40.5 mol% of G + C content. Based on these results, we propose a new species of the genus Streptococcus, for which the name Streptococcus thalassemiae sp. nov., Marseille-Q2617T (=CSUR Q2617 = CECT 30109) was proposed.

Bartonella raoultii sp. nov., isolated from infected rodents (Mastomys erythroleucus) in Senegal.

Bartonella species are involved in various human diseases, causing a range of clinical manifestations; animals are considered as the main reservoirs, transmitting diverse species of Bartonella through direct contact and haematophagous insects. Here, we characterize a new species, Bartonella raoultii sp. nov., within the genus Bartonella, using a taxonogenomic polyphasic approach. Strain 094T (= CSUR B1097T=DSM 28004T), isolated from the blood of an infected rodent (Mastomys erythroleucus) in Senegal, is an aerobic and rod-shaped bacterium. The annotated non-contiguous genome sequence is 1 952322 bp long and contains 37.2 mol% G+C content, 1686 protein-coding genes and 50 RNA genes, including seven rRNA genes.

Hymenopteran Parasitoids of Hard Ticks in Western Africa and the Russian Far East.

Some parasitoids of the genus Ixodiphagus (Hymenoptera, Chalcidoidea: Encyrtidae) are well-known natural enemies of ticks. In this study, we investigate the occurrence of parasitoid wasps in adult hard ticks from Western Africa (Côte d'Ivoire and Senegal) and Far Eastern Europe (Russia) using molecular methods. The morphological identification allowed the classification of 785 collected specimens of six species of ticks: Rhipicephalus (Boophilus) microplus (41%), Ixodes persulcatus (33%), Dermacentor silvarum (11%), Haemaphysalis concinna (7%), Amblyomma variegatum (5%), and Haemaphysalis japonica (3%). The newly developed MALDI-TOF MS protocol identified tick species in spite of their different storage (dried or in 70% ethanol) conditions for a long period. Molecular screening of ticks by a new standard PCR system developed in this study revealed the presence of parasitoid wasp DNA in 3% (28/785) of analyzed ticks. Ixodiphagus hookeri was detected in 86% (24/28) of infested ticks, including 13 I. persulcatus, 9 R (B) microplus, and one H. concinna and D. silvarum. While an unidentified parasitoid wasp species from the subfamily Aphidiinae and Braconidae family was detected in the remaining 14% (4/28) infested ticks. These infested ticks were identified as I. persulcatus. Our findings highlight the need for further studies to clarify the species diversity of parasitoid infesting ticks by combining molecular and morphological features. The novel molecular and MALDI-TOF MS protocols could be effective tools for the surveillance and characterization of these potential bio-control agents of ticks.