SARS-CoV-2 antibodies seroprevalence in dogs from France using ELISA and an automated western blotting assay.

Dogs are occasionally susceptible to SARS-CoV-2, developing few or no clinical signs. Epidemiological surveillance of SARS-CoV-2 in dogs requires testing to distinguish it from other canine coronaviruses. In the last year, significant advances have been made in the diagnosis of SARS-CoV-2, allowing its surveillance in both human and animal populations. Here, using ELISA and automated western blotting (AWB) assays, we performed a longitudinal study on 809 apparently healthy dogs from different regions of France to investigate anti-SARS-CoV-2 antibodies. There were three main groups: (i) 356 dogs sampled once before the pandemic, (ii) 235 dogs sampled once during the pandemic, and (iii) 218 dogs, including 82 dogs sampled twice (before and during the pandemic), 125 dogs sampled twice during the pandemic and 11 dogs sampled three times (once before and twice during the pandemic). Using ELISA, seroprevalence was significantly higher during the pandemic [5.5% (25/453)] than during the pre-pandemic period [1.1% (5/449)]. Among the 218 dogs sampled twice, at least 8 ELISA-seroconversions were observed. ELISA positive pre-pandemic sera were not confirmed in serial tests by AWB, indicating possible ELISA cross-reactivity, probably with other canine coronaviruses. A significant difference was observed between these two serological tests (Q = 88, p = 0.008). A clear correlation was observed between SARS-CoV-2 seroprevalence in dogs and the incidence of SARS-CoV-2 infection in human population from the same area. AWB could be used as a second line assay to confirm the doubtful and discrepant ELISA results in dogs. Our results confirm the previous experimental models regarding the susceptibility of dogs to SARS-CoV-2, suggesting that viral transmission from and between dogs is weak or absent. However, the new variants with multiple mutations could adapt to dogs; this hypothesis cannot be ruled out in the absence of genomic data on SARS-CoV-2 from dogs.

Great diversity of Piroplasmida in Equidae in Africa and Europe, including potential new species.

Piroplasms are Apicomplexa tick-borne parasites distributed worldwide. They are responsible for piroplasmosis (theileriosis and babesiosis) in Vertebrata and are therefore of medical and economic importance. Herein, we developed a new real time PCR assay targeting the 5.8S rRNA gene and three standard PCR assays, targeting 18S rRNA, 28S rRNA, and cox1 genes, for the detection of piroplasmids. These assays were first optimized and screened for specificity and sensitivity. Then, they were used to study a total of 548 blood samples and 97 ticks collected from Equidae in four sub-Saharan countries (Senegal, Democratic Republic of the Congo, Chad, and Djibouti) and France (Marseille and Corsica). DNA of piroplasms was detected in 162 of 548 (29.5%) blood samples and in 9 of 97 (9.3%) ticks. The highest prevalence in blood samples was observed in Chad in 2016 with 72.9% positivity rate. Sequencing allowed the identification of four species of piroplasms, including two potentials new species. Theileria equi was mainly found. The highest prevalence was observed in Senegal (14 positive out of 23, 60.87%). Babesia caballi was detected in one horse in Senegal. Two new potential Theileria species were detected: Theileria sp. "Africa", observed in all areas excepted in Marseille and Theileria sp. "Europa", observed in Marseille and Corsica. In conclusion, sensitive and specific PCR assays were developed for epidemiological studies of Piroplasmida. The circulation of multiple species of piroplasms, including two potentials new species, observed among Equidae from sub-Saharan Africa and France.

Molecular investigation and phylogeny of species of the Anaplasmataceae infecting animals and ticks in Senegal.

BACKGROUND: Our study aimed to assess the diversity of the species of Anaplasmataceae in Senegal that infect animals and ticks in three areas: near Keur Momar Sarr (northern region), Dielmo and Diop (Sine Saloum, central region of Senegal), and in Casamance (southern region of Senegal). METHODS: A total of 204 ticks and 433 blood samples were collected from ruminants, horses, donkeys and dogs. Ticks were identified morphologically and by molecular characterization targeting the 12S rRNA gene. Molecular characterization of species of Anaplasmataceae infecting Senegalese ticks and animals was conducted using the 23S rRNA, 16S rRNA, rpoB and groEL genes. RESULTS: Ticks were identified as Rhipicephalus evertsi evertsi (84.3%), Hyalomma rufipes (8.3%), Hyalomma impeltatum (4.9%), R. bursa (1.5%) and R. muhsamae (0.9%). The overall prevalence of Anaplasmataceae infection in ticks was 0.9%, whereas 41.1% of the sampled animals were found infected by one of the species belonging to this family. We identified the pathogen Anaplasma ovis in 55.9% of sheep, A. marginale and A. centrale in 19.4% and 8.1%, respectively, of cattle, as well as a putative new species of Anaplasmataceae. Two Anaplasma species commonly infecting ruminants were identified. Anaplasma cf. platys, closely related to A. platys was identified in 19.8% of sheep, 27.7% of goats and 22.6% of cattle, whereas a putative new species, named here provisionally "Candidatus Anaplasma africae", was identified in 3.7% of sheep, 10.3% of goats and 8.1% of cattle. Ehrlichia canis and Anaplasma platys were identified only from dogs sampled in the Keur Momar Sarr area. Ehrlichia canis was identified in 18.8% of dogs and two R. e. evertsi ticks removed from the same sheep. Anaplasma platys was identified in 15.6% of dogs. Neither of the dogs sampled from Casamance region nor the horses and donkeys sampled from Keur Momar Sarr area were found infected by an Anaplasmataceae species. CONCLUSIONS: This study presents a summary of Anaplasmataceae species that infect animals and ticks in three areas from the northern, central and southern regions of Senegal. To our knowledge, our findings demonstrate for the first time the presence of multiple Anaplasmataceae species that infect ticks and domestic animals in Senegal. We recorded two potentially new species commonly infecting ruminants named here provisionally as Anaplasma cf. platys and "Candidatus Anaplasma africae". However, E. canis was the only species identified and amplified from ticks. None of the other Anaplasmataceae species identified in animals were identified in the tick species collected from animals.

Leptospirosis, one neglected disease in rural Senegal.

A serological study was carried out in two Senegalese villages located in the Sine-Saloum region in order to estimate the presence of anti-leptospiral antibodies in humans and animals, and to identify the predominant serogroups. Seven hundred and forty-nine serum samples were collected from humans (n = 545), dogs (n = 33), donkeys (n = 20), goats (n = 52), sheep (n = 43) and N'Dama cattle (n = 56), all originated from Dielmo and Ndiop villages. All samples were tested for different serovars of pathogenic Leptospira species by the microscopic agglutination test. Considering titres ≥ 1:100, 7.7% [CI 95:5.5 to 9.9] on the 545 human blood samples tested and 42.2% [CI95 :35.4 to 48.9] on the 204 animal blood samples tested were found to be positive to one or more serovars. The results obtained indicate that the Australis serogroup is the most prevalent serogroup in human (67.3%) and cattle (27.3%). Serogroup Icterohaemorhagiae is the most frequent serogroup in goat (55.6%) and donkey (37.5%). Canicola (23.4%), Icterohaemorhagiae (21.1%) and Australis (12.5%) serogroups are the most prevalent serogroups in dogs. This study shows that diverse Leptospira serovars occur in a wide range of wild and domestic mammal species, as well as in humans in Senegal. However, further studies are needed to better understand the complexity of Leptospira epidemiology in Africa, identify the reservoirs of different serogroups and estimate its impact on livestock. Understanding the multi-host epidemiology of leptospirosis is essential to control and prevent the disease.

Bartonella bovis and Candidatus Bartonella davousti in cattle from Senegal.

In Senegal, domestic ruminants play a vital role in the economy and agriculture and as a food source for people. Bartonellosis in animals is a neglected disease in the tropical regions, and little information is available about the occurrence of this disease in African ruminants. Human bartonellosis due to Bartonella quintana has been previously reported in Senegal. In this study, 199 domestic ruminants, including 104 cattle, 43 sheep, and 52 goats were sampled in villages from the Senegalese regions of Sine Saloum and Casamance. We isolated 29 Bartonella strains, all exclusively from cattle. Molecular and genetic characterization of isolated strains identified 27 strains as Bartonella bovis and two strains as potentially new species. The strains described here represent the first Bartonella strains isolated from domestic ruminants in Senegal and the first putative new Bartonella sp. isolated from cattle in Africa.

Study of ehrlichiosis in kennel dogs under treatment and prevention during seven months in Dakar (Senegal).

In Dakar kennels where morbidity and mortality attributed to diseases transmitted by ticks were high, we conducted a field study to assess the prevalence of Ehrlichia canis, Anaplasma platys and Babesia spp. infections in two kennels (n = 34 dogs) and to study the impact of tick protection. The first day of the study, the E. canis PCR were positive in 18 dogs (53%). A. platys was found in one dog and all dogs were negative for Babesia spp. After one month of doxycycline treatment, the number of PCR positive dogs decreased significantly to 2 (5.9%). During seven months, all dogs were treated monthly topically with a novel combination (Certifect(®), Merial) delivering at least 6.7 mg fipronil/kg body weight, 8.0mg amitraz/kg and 6 mg (S)-methoprene/kg. The number of PCR positive dogs remained stable all over the seven months, with 4 dogs being positive at Day 90 and 2 at Day 210. The combination of treatment and monthly prevention had a significant effect in the two kennels. All dogs remained healthy, which was not the case in previous years.