Surveillance of Fleas and Their Small Mammal Hosts for Plague Risks in Some Main Seaports of the Islands of the Southwestern Indian Ocean.

Since ancient times, seaports have been the hot spots for plague introduction into free countries. Infected ship rats reached new areas, and epizootics occurred prior to human infection via flea bites. Beginning in the 1920s/1930s, rodent and flea surveillance was carried out as part of plague hazard management in seaports of the world. Nowadays, such activity is not done regularly. In the southwestern Indian Ocean (SWIO) region, plague surveillance is of great importance given plague endemicity in Madagascar and thus the incurred risk for neighboring islands. This study reports animal-based surveillance aimed at identifying fleas and their small mammal hosts in SWIO seaports as well as Yersinia pestis detection. Small mammal trappings were performed in five main seaports of Madagascar (Toamasina and Mahajanga), Mauritius (Port Louis), and the Union of Comoros (Moroni and Mutsamudu). Mammals were euthanized and their fleas collected and morphologically identified before Y. pestis detection. In total, 145 mammals were trapped: the brown rat Rattus norvegicus (76.5%), the black rat Rattus rattus (8.3%), and the Asian house shrew Suncus murinus (15.2%). Fur brushing allowed collection of 1,596 fleas exclusively identified as Xenopsylla cheopis. All tested fleas were negative for Y. pestis DNA. This study shows that both well-known plague mammal hosts and flea vectors occur in SWIO seaports. It also highlights the necessity of carrying out regular animal-based surveillance for plague hazard management in this region.

First report of Fusarium wilt of Cavendish bananas caused by Fusarium oxysporum f. sp. cubense Tropical Race 4 in the Grande Comoros Island.

Fusarium wilt of banana, caused by the soil-borne pathogen Fusarium oxysporum f. sp. cubense (Foc), is a major constraint to banana production worldwide (Viljoen et al., 2020). Currently, Cavendish bananas are severely affected by Foc Tropical Race 4 (TR4) globally. In Africa, Foc TR4 was first detected in northern Mozambique in 2013 (Viljoen et al., 2020), and has since been found on the island of Mayotte in the Mozambique Channel off the coast of southeastern Africa (Aguayo et al., 2021). In early 2023, severe leaf-yellowing and wilting of Cavendish banana plants was observed on Cavendish banana plants at several small holder farmer properties in Grande Comoros (Ngazidja) including in Ntsinimoipanga (-11,790054 S, 43°25'47,04384 E), Batou (-11,499716 S, 43°21'51,71976''E), Madjeweni (-11,8217 S , 43°16'41''E) and Mdé (-11°41'54'' S, 43°15'20''E). When the pseudostems of these plants were split open, a reddish-brown internal discoloration of the vascular tissue became apparent. Discolored strands of diseased plants were collected, and the causal agent identified using DNA-based techniques, vegetative compatibility group (VCG) analysis and pathogenicity testing. The samples were plated onto potato dextrose agar and single-spored and isolated from individual diseased plants identified as F. oxysporum based on cultural and morphological characteristics. These include the production of white fungal colonies with a purple center, infrequent production of macroconidia, but an abundance of microconidia on short monophialides, and terminal or intercalary chlamydospores (Leslie and Summerell, 2006). Foc TR4 was identified from seven isolates by conventional (Dita et al., 2010) and quantitative-PCR (Matthews et al., 2020), and with loop-mediated isothermal amplification (LAMP) (Ordóñez et al., 2021). All seven isolates were confirmed as members of the VCG 01213/16 complex when nit-1 mutants of the unknown Foc isolates were compatible with Nit-M mutants of the Foc VCG 01213 and VCG 01216 tester strains. Two isolates were then selected for pathogenicity testing, and 2-month-old tissue culture-derived Cavendish plants (cv. Williams) inoculated by using the method described by Ndayihanzamaso et al. (2022). After 4 weeks, the Foc TR4-inoculated plants produced wilting symptoms and internal rhizome discoloration typical of Fusarium wilt. Foc TR4 was reisolated from the inoculated plants and identified by qPCR (Matthews et al. 2020), thereby fulfilling Koch's postulates. These results provide scientific proof of the presence of Foc TR4 in a second island in the Comoros archipelago. Comprehensive surveys will be conducted in all three of the Comoros Islands to assess the presence and impact of Foc TR4 to implement containment strategies. Collaborative initiatives and coordinated actions among growers and other stakeholders are needed to prevent the spread of Foc TR4 to more Southwest Indian Ocean islands and countries on the East African coasts. The importance of banana for food security and livelihoods, and the unique genetic diversity of bananas found on the Comoros islands, requires the eradication and isolation of diseased bananas on the short term, and the screening of local banana varieties for Foc TR4 resistance on the longer term.

Large-scale mitochondrial DNA analysis of native honey bee Apis mellifera populations reveals a new African subgroup private to the South West Indian Ocean islands.

BACKGROUND: The South West Indian Ocean (SWIO) archipelagos and Madagascar constitute a hotspot of biodiversity with a high rate of endemism. In this area, the endemic subspecies A. m. unicolor has been described in Madagascar. It belongs to the African lineage, one of the four described evolutionary lineages in honey bees. Despite a long beekeeping tradition and several recorded European introductions, few studies have been carried out on the diversity and proportion of honey bee subspecies. In order to identify and define which evolutionary lineages and potential sub-lineages are present in the SWIO, the COI-COII intergenic region and the ND2 gene of the mtDNA were sequenced in honey bee colonies from three archipelagos. An extensive sampling (n = 1184 colonies) was done in the Mascarene (La Réunion, Mauritius, Rodrigues), Seychelles (Mahé, Praslin, La Digue) and Comoros (Grande Comore, Mohéli, Anjouan, Mayotte) archipelagos. Islands genetic diversity was compared to newly sampled populations from Madagascar, continental African and European populations. RESULTS: African lineage haplotypes were found in all islands (except for Rodrigues). Madagascar, Comoros and Seychelles had 100% of A lineage, 95.5% in La Réunion and 56.1% in Mauritius. Among all African colonies detected in the SWIO, 98.1% (n = 633) of COI-COII haplotypes described the presence of the subspecies A. M. unicolor. Both genetic markers revealed i) a new private AI mitochondrial group shared by the SWIO archipelagos and Madagascar distant from continental populations; ii) the private African haplotypes for each island suggested diversity radiation in the archipelagos; iii) the detection of the Comoros archipelago as a possible contact area between insular and continental African populations. The exotic European C and M lineages were only detected in the Mascarene archipelago, but striking differences of proportion were observed among islands. Merely 4.6% of European colonies were found in La Réunion whereas Mauritius cumulated 44%. Here, among the 84 observed COI-COII haplotypes, 50 were newly described including 13 which were private to the SWIO archipelagos and Madagascar. Similarly, 24 of the 34 found ND2 haplotypes were novel which included six haplotypes particular to the SWIO populations. CONCLUSION: A new African subgroup was described in the SWIO region with mitochondrial genetic evidence that A. m. unicolor is the indigenous subspecies of the archipelagos surrounding Madagascar.