Molecular Epidemiology of Peste Des Petits Ruminants Virus in West Africa: Is Lineage IV Replacing Lineage II in Burkina Faso?

This study aimed at investigating the genetic lineages of peste des petits ruminants virus (PPRV) currently circulating in Burkina Faso. As part of PPR surveillance in 2021 and 2022, suspected outbreaks in different regions were investigated. A risk map was produced to determine high-risk areas for PPR transmission. Based on alerts, samples were obtained from three regions and all sampled localities were confirmed to fall within PPR high risk areas. We collected swab samples from the eyes, mouth, and nose of sick goats. Some tissue samples were also collected from dead animals suspected to be infected by PPRV. In total, samples from 28 goats were analysed. Virus confirmation was performed with RT-PCR amplification targeting the nucleocapsid (N) gene. Partial N gene sequencing (350 bp) was carried out using the RT-PCR products of positives samples to characterise the circulating lineages. Eleven sequences, including ten new sequences, have been obtained. Our study identified the presence of the PPRV lineage IV in the three studied regions in Burkina Faso with a genetic heterogeneity recorded for the sequences analysed. Previously published data and results of this study suggest that PPRV lineage IV seems to be replacing lineage II in Burkina Faso.

Evaluation of synergism in essential oils against the cattle tick Rhipicephalus microplus in Burkina Faso.

The cattle tick Rhipicephalus microplus affects animal production economically by reducing weight gain and milk production and causing diseases, such as babesiosis and anaplasmosis. Using synthetic acaricides to reduce their incidence has caused the emergence of resistant tick populations. The present study aimed to assess the in vitro acaricidal activity of combinations of essential oils (EOs) from Ocimum americanum, Ocimum gratissimum, and Lippia multiflora against R. microplus larvae. In fact, numerous biological properties have been reported on EOs from these three plants, including acaricidal properties. Hence, a larval immersion test was performed using a population of R. microplus resistant to synthetic acaricides used in Burkina Faso. Results revealed that EO from O. gratissimum was the most effective on R. microplus larvae with LC50 and LC90 values at 10.36 and 15.51 mg/mL, respectively. For EO combinations, the most significant synergistic effect was obtained by combination 6 (1/3 O. americanum + 2/3 O. gratissimum +1/6 L. multiflora), with a combination index value of 0.44. All combinations presented dose reduction index >1, indicating a favorable dose reduction. According to the literature, this is the first study to determine the combination effect of EOs from the abovementioned plants in controlling R. microplus activity in vitro. Thus, the combination of these EOs is an alternative to control the resistant populations of invasive cattle ticks.

Efficacy of local essential oils against Amblyomma variegatum tick from Burkina Faso.

The native tick Amblyomma variegatum remains one of the most important tick species affecting cattle in West Africa. This hinders animal production by negatively impacting the health and reproduction of animals infested with the tick. Given the negative consequences on production quality and environmental health, the use of chemical products for tick control is increasingly being discouraged. Therefore, this study aimed to assess the acaricidal activity of essential oils such as Ageratum conyzoïdes, Cymbopogon citratus, Cymbopogon giganteus, Lippia multiflora and Ocimum gratissimum against specimens of A. variegatum from Burkina Faso. A larval immersion test was performed to investigate the larvicidal activities of these essential oils. Gas chromatography-mass spectrometry was used to determine the chemical compositions of essential oils. The chemical composition was predominantly oxygenated monoterpenes in A. conyzoïdes (48.71 %), C. citratus (99.9 %) and C. giganteus (73.63 %), while hydrocarbon monoterpenes were the most abundant in O. gratissimum (63.7 %) and hydrocarbon sesquiterpenes in L. multiflora (71.719 %). The recorded larvicidal activity, varied according to the species of plants and the dose applied. At a dose of 12.5 mg/mL, all essential oils studied, except L. multiflora (7.54 %), induced 100 % larval mortality. In this study, we highlight the promising larvicidal effects of local essential oils against A. variegatum. These essential oils can be used as bio-acaricides, which are effective and environmentally-friendly alternatives to chemical products. However, further investigations are required to determine the mechanisms of action of these essential oils for in vivo experimentation and their practical application in the control of A. variegatum ticks.

The potential of Rhipicephalus microplus as a vector of Ehrlichia ruminantium in West Africa.

Heartwater, or cowdriosis, is a virulent tick-borne rickettsial disease of ruminants caused by Ehrlichia ruminantium, biologically transmitted by Amblyomma species (A. variegatum in West Africa). In West Africa, this bacterium was recently reported to naturally infect the invasive cattle tick, Rhipicephalus microplus (Rm) through trans-ovarian transmission from replete adult females to offspring. A 'sheep-tick-sheep' cycle was set up to determine whether feeding the progeny of these ticks on naïve sheep could lead to infection, and to compare clinical outcomes resulting from this transmission with those observed following infection by the natural A. variegatum (Av) vector. Using local strains of ticks (KIMINI-Rm and KIMINI-Av) and of E. ruminantium (BK242), we recorded, using the PCR technique, the presence of bacterial DNA in ticks (larvae for Av and females for Rm) engorged on sheep inoculated by BK242-infected blood. The bacterial DNA was also detected in the next stages of the lifecycle of R. microplus (eggs and larvae), and in sheep infested either by those R. microplus larvae or by A. variegatum nymphs moulted from larvae engorged on blood-inoculated sheep. Bacterial infection in these sheep was demonstrated by detecting antibodies to E. ruminantium using the MAP1-B ELISA and by isolation of the bacterium on cell culture from blood. The sequences of PCS20 gene detected in ticks and sheep were identical to that of the BK242 strain. Our results confirm that R. microplus can acquire and transmit E. ruminantium to the next stage. However, this transmission resulted in a mild subclinical disease whereas severe clinical disease was observed in sheep infested by A. variegatum infected nymphs, suggesting differences in the tick/bacteria relationship. Future studies will focus on replicating these findings with ticks of different isolates and life stages to determine if R. microplus is playing a role in the epidemiology of heartwater in West Africa. Additionally, studies will investigate whether sheep that are seropositive due to infestation by E. ruminantium-infected R. microplus are subsequently protected against heartwater. Such data will add to our understanding of the possible impact of R. microplus in areas where it has become recently established.

New geographical records for tick-borne pathogens in ticks collected from cattle in Benin and Togo.

BACKGROUND: Ticks are obligate hematophagous arthropods capable of transmitting a great variety of endemic and emerging pathogens causing diseases in animals and humans. OBJECTIVES: The aim of this study was to investigate the presence of Bartonella spp., Rickettsia spp., Borrelia burgdorferi sensu lato (s.l.) and Anaplasma phagocytophilum in ticks collected from cattle in Benin and Togo. METHODS: Overall, 396 (148 males, 205 females and 43 nymphs) ticks were collected from cattle in 17 districts (Benin and Togo) between 2019 and 2020. Ticks were pooled into groups of 2-6 ticks per pool according to individual host, location, species and developmental stage. The DNA of each pool was extracted for molecular screening. RESULTS: PCR results revealed that 20 tick pools were positive for Bartonella spp. (Benin and Togo) and 23 tick pools positive for Rickettsia spp. (Benin), while all pools were negative for A. phagocytophilum and B. burgdorferi s.l. Sequence analysis of positive Rickettsia samples revealed the presence of Rickettsia aeschlimannii. CONCLUSIONS: The present study highlights the presence of zoonotic agents in ticks collected from cattle in Benin and Togo. This information will raise awareness of tick-borne diseases among physicians and veterinarians, stimulate further studies to monitor these pathogens, and advise on necessary measures to control the spread of these zoonoses.

Tick species infesting cattle in the central region of Burkina Faso: Presence of Rhipicephalus microplus less than ten years after its first identification in the Southwest part of the country.

Ticks are among the most important vectors for medical and veterinary importance in African countries, representing thus an obstacle for development and improvement of livestock production. The central part of Burkina Faso is an arid region which has a low diversity of tick species and in which Rhipicephalus microplus has been absent. This study aimed to examine ticks infesting cattle within this region, focusing on the peripheral area of Ouagadougou. Tick sampling was undertaken monthly on 30 selected cattle in the state breeding station of Loumbila, from June 2017 to February 2018. Morphological identification was carried out based on previous established keys and catalogue. A total of 1,076 ticks belonging to four genera and seven species were collected. Amblyomma variegatum (50.5%) was the most abundant species followed by Hyalomma rufipes (27.4%), Hyalomma truncatum (13.7%), Rhipicephalus decoloratus (4.5%), Hyalomma impressum (2.4%), Rhipicephalus microplus (1.5%), and Rhipicephalus sanguineus sensu lato (0.1%). The results suggested a decrease of A. variegatum and Hyalomma spp. load compared to data collected in 1996 from the same study area. Presence of R. microplus and R. decoloratus has been highlighted in the studied area for the first time. This finding highlights the relevance of regular investigations on tick species infesting livestock utilizing collaboration with different stakeholders.

Modelling habitat suitability of the invasive tick Rhipicephalus microplus in West Africa.

Ticks have medical and economic importance due to their ability to transmit pathogens to humans and animals. In tropical and sub-tropical countries, tick-borne diseases (TBD) are among the most important diseases affecting livestock and humans. The fast spread of ticks and TBD requires a quick development and application of efficient prevention and/or control programs. Therefore, prior investigations on TBD and related vectors epidemiology, for instance, through accurate epidemiological models, are mandatory. This study aims to develop models to forecast suitable habitat for Rhipicephalus microplus distribution in West Africa. Tick occurrences were assembled from 10 different studies carried out in six West African countries in the past decade. Six statistical models (maximum entropy in a single model and generalised linear model, generalised additive model, random forest, boosted regression tree and support vector machine model in an ensemble model) were applied and compared to predict the habitat suitability of R. microplus distribution in West Africa. Each model was evaluated with the area under the receiver operating characteristic curve (AUC), the true skill statistic (TSS) and the Boyce index (BI). The selected models had good performance according to their AUC (above .8), TSS (above .7) and BI (above .8). Temperature played a key role in MaxEnt model, whereas normalised difference vegetation index (NDVI) was the most important variable in the ensemble model. The model predictions showed coastal countries of West Africa as more suitable for R. microplus. However, some Sahelian areas seems also favourable. We stress the importance of vector surveillance and control in countries that have not yet detected R. microplus but are in the areas predicted to host suitable habitat. Indeed, awareness-raising and training of different stakeholders must be reinforced for better prevention and control of this tick in these different countries according to their status.

Models for Studying the Distribution of Ticks and Tick-Borne Diseases in Animals: A Systematic Review and a Meta-Analysis with a Focus on Africa.

Ticks and tick-borne diseases (TTBD) are constraints to the development of livestock and induce potential human health problems. The worldwide distribution of ticks is not homogenous. Some places are ecologically suitable for ticks but they are not introduced in these areas yet. The absence or low density of hosts is a factor affecting the dissemination of the parasite. To understand the process of introduction and spread of TTBD in different areas, and forecast their presence, scientists developed different models (e.g., predictive models and explicative models). This study aimed to identify models developed by researchers to analyze the TTBD distribution and to assess the performance of these various models with a meta-analysis. A literature search was implemented with PRISMA protocol in two online databases (Scopus and PubMed). The selected articles were classified according to country, type of models and the objective of the modeling. Sensitivity, specificity and accuracy available data of these models were used to evaluate their performance using a meta-analysis. One hundred studies were identified in which seven tick genera were modeled, with Ixodes the most frequently modeled. Additionally, 13 genera of tick-borne pathogens were also modeled, with Borrelia the most frequently modeled. Twenty-three different models were identified and the most frequently used are the generalized linear model representing 26.67% and the maximum entropy model representing 24.17%. A focus on TTBD modeling in Africa showed that, respectively, genus Rhipicephalus and Theileria parva were the most modeled. A meta-analysis on the quality of 20 models revealed that maximum entropy, linear discriminant analysis, and the ecological niche factor analysis models had, respectively, the highest sensitivity, specificity, and area under the curve effect size among all the selected models. Modeling TTBD is highly relevant for predicting their distribution and preventing their adverse effect on animal and human health and the economy. Related results of such analyses are useful to build prevention and/or control programs by veterinary and public health authorities.

Cross border transhumance involvement in ticks and tick-borne pathogens dissemination and first evidence of Anaplasma centrale in Burkina Faso.

In West Africa, cross-border transhumance, also called seasonal migration, is known to be a very important animal production strategy, as it involves about 70 to 90% of cattle. In spite of the cattle movements, some strategic areas of transhumance remain poorly explored regarding ticks and their associated pathogens investigations. The purpose of this study is to evaluate the involvement of transhumance in the spread of cattle ticks and associated pathogens in Burkina Faso (BF) and Benin (BN), in a context of speedy invasion of West African livestock by Rhipicephalus microplus. A longitudinal survey was performed on 210 cattle from BF, monitored for ticks and tick-borne pathogens (TBP) during one seasonal transhumance. The first sampling coded "T0BF" took place in eastern BF, at the transhumance departure. A second sampling "T1BN" was carried out in northern BN, the transhumance arrival zone. A third sampling "T2BF" was done at the return of cattle in eastern BF. Ticks were morphologically identified and TBP detected with reverse line blot hybridization (RLB) assay. A total of 1027 ticks (7 species), 1006 ticks (11 species) and 1211 ticks (9 species) were respectively found at T0BF, T1BN and T2BF. Some species were collected at the three times of sampling without any significant difference in their relative abundances. However, other tick species appeared only at T1BN and/or T2BF. The TBP species found at the three points surveyed were Theileria annulata, Theileria mutans, Theileria velifera, Babesia bigemina and Anaplasma marginale. The most prevalent was T. mutans with 166/210 (79%), 159/210 (75.7%) and 78/210 (37%) cattle positive respectively at T0BF, T1BN and T2BF. Anaplasma centrale was evidenced with 0.5% and 0.9% respectively at T0BF and T2BF. To our knowledge, this represents its first report in the study area. Overall, the TBP prevalences were significantly lower at T2BF, highlighting the effect of tick populations changes induced by transhumance combined with the seasonal variation influence.

Efficacy of two commercial synthetic pyrethroids (cypermethrin and deltamethrin) on Amblyomma variegatum and Rhipicephalus microplus strains of the south-western region of Burkina Faso.

Since 2011, period of the livestock invasion by the cattle tick Rhipicephalus microplus in Burkina Faso (BF), tick-control problems were exacerbated. Based on farmer's reports, most commonly used commercial acaricides were found to be ineffective in Western South part of the country. To investigate the occurrence and extent of such acaricidal ineffectiveness, we performed the standardized larval packet test (LPT) with commercial deltamethrin (vectocid) and cypermethrin (cypertop), on two cattle tick species, the native Amblyomma variegatum and the invasive R. microplus. The resistance ratios (RR) were computed with susceptible Hounde strain of Rhipicephalus geigyi as reference. The R. microplus population showed resistance to the two acaricides tested with the highest lethal concentration (LC) values, and different resistance ratios higher than 4 (deltamethrin: RR50 = 28.18 and RR90 = 32.41; cypermethrin: RR50 = 8.79 and RR90 = 23.15). In the contrary, A. variegatum population was found to be highly susceptible to acaricides tested with low lethal concentrations and resistance ratio values (deltamethrin: RR50 = 0.5 and RR90 = 0.48; cypermethrin: RR50 = 0.68 and RR90 = 0.79). These data demonstrate high synthetic pyrethroid resistance in R. microplus strain, leading to conclude that the acaricide ineffectiveness in tick populations control remains a concern in BF.

First tick and tick damage perception survey among sedentary and transhumant pastoralists in Burkina Faso and Benin.

BACKGROUND: Transhumance, a main ancestral animal production strategy of the West African Countries (WAC), can favour the spread of vectors and vector-borne diseases within and/or across countries. Transhumance has been implicated in such spread as well as that of related tick-borne diseases (TBD). METHODS AND PRINCIPAL FINDINGS: Using a questionnaire survey and statistical modelling, this study explores the perception of herders about ticks and TBD in cattle, their practices in tick control and the social groups involved in cattle farming in eastern Burkina Faso (46 random herds) and in the northern Benin (44 random herds). Results show that most of the herders (79%) are from the Fulani social group. The principal and secondary activities of herders are respectively cattle farming and agriculture. The mean age of pastoralists is between 40 and 50 years depending on the province of origin and 60% of the surveyed herds practice internal or transboundary transhumance. Herders have a clear knowledge of different genus of ticks except the genus Rhipicephalus. Their knowledge of TBD is very limited. These results also reveal that herders in Benin use less acaricides treatment calendar compared with those in Burkina Faso. Transhumant pastoralists (i.e. transhumant cattle farmers) plan more acaricide treatment and have more cows with lost teats (i.e. tick damage) than the sedentary ones. In addition, amitraz appears to be the main acaricide compound used by herders for tick control (68%) but its use is inappropriate and its source is frequently the unregulated market. CONCLUSIONS AND SIGNIFICANCE: All of these findings can induce acaricide resistance especially as the inefficacy of amitraz against Rhipicephalus (Boophilus) microplus has already been reported in previous studies. Such results would help to elaborate suitable strategies of control and prevention of ticks and TBD in Burkina Faso and Benin.

Cattle ticks and associated tick-borne pathogens in Burkina Faso and Benin: Apparent northern spread of Rhipicephalus microplus in Benin and first evidence of Theileria velifera and Theileria annulata.

Babesiosis, theileriosis, anaplasmosis, and heartwater are tick-borne diseases that threaten livestock production in sub-Saharan Africa including Burkina Faso and Benin. For over a decade, these two bordering countries have been facing an invasion of the livestock by the tick Rhipicephalus microplus, a major vector for babesiosis, accidentally introduced in Benin in 2004. The molecular identification of tick-borne pathogens in this border area is of particular interest due to animals seasonal migration between the two countries. In this survey, epidemiological features of ticks and tick-borne pathogens in cattle were investigated to compare the eastern Burkina Faso, corresponding to a seasonal migration departure zone, and the northern Benin, which represents a seasonal migration arrival zone. Ticks and peripheral blood were collected from a total of 946 cattle in the two areas. Ticks were morphologically identified and the DNA samples from bovine blood and ticks were analysed by Reverse Line Blot (RLB) hybridization process. A total of 2856 ticks were collected on 490 cattle in Burkina Faso, eight tick species were identified, while 3583 ticks were collected on 456 cattle in North Benin with nine tick species identified. The invasive tick, R. microplus was not found in eastern Burkina Faso, but its spread farthest north in Benin is reported. Six tick-borne pathogen species were found in cattle blood both in eastern Burkina Faso and in northern Benin. Ranked in decreasing order of overall prevalences, they are: Theileria mutans (91.1%), Theileria velifera (77.8%), Babesia bigemina (10.9%), Anaplasma marginale (4.2%), Babesia bovis (3.3%), and Theileria annulata (1.8%). To the best of our knowledge, this survey represents the first report of T. velifera and T. annulata in the region. Overall, the TBP prevalences were significantly higher in northern Benin than in eastern Burkina Faso, indicating a higher parasitological risk in this area.

First digital characterization of the transhumance corridors through Benin used by cattle herds from Burkina Faso and associated risk scoring regarding the invasion of Rhipicephalus (Boophilus) microplus.

BACKGROUND: Livestock plays a key role in the macro-economy of West Africa and provides livelihoods for millions of people. The main cattle rearing strategy in West Africa is pastoralism, including transhumance, that is a seasonal migration of cattle with their herders. This adaptive strategy aims to optimize livestock access to water and pastures. However, it can favour pathogens and vectors transboundary spread. The aim of this study was to highlight firstly, the corridors and grazing areas used by Burkina Faso transhumant cattle herds going to Benin; secondly, the characteristics of departure and arrival areas of transhumance; and thirdly, the risk score related to introduction and spread of the invasive tick species, Rhipicephalus (Boophilus) microplus, in free areas. METHODS AND PRINCIPAL FINDINGS: Therefore, GPS devices were given to 27 herders to monitor a full transhumance season between East Burkina Faso and North Benin. The analysis of 14,966 spots generated by the GPS devices revealed four main corridors and five main grazing areas used by cattle herds during transhumance. Statistical analysis of normalized difference vegetation index (NDVI), rainfall and temperature data, highlighted significant differences between departure and arrival areas. NDVI and rainfall are significantly higher in Benin than Burkina Faso, whereas temperature is significantly lower. Additionally, using biotic and abiotic parameters, a risk scoring was developed to predict the presence of Rhipicephalus (Boophilus) microplus at municipality level. CONCLUSIONS AND SIGNIFICANCE: The better vegetation, temperature and rainfall conditions during the dry seasons in Benin attract cattle herds from Burkina Faso. The invasiveness and adaptability of Rhipicephalus (Boophilus) microplus added to the frequent stays of transhumant herds in infested areas suggest its potential introduction and establishment in free areas soon. Moreover, frequent intrusions of the transhumant cattle in the wildlife reserves are another risk of vectors and pathogen exchange between domestic and wild animals.

The Recent Evolution of a Maternally-Inherited Endosymbiont of Ticks Led to the Emergence of the Q Fever Pathogen, Coxiella burnetii.

Q fever is a highly infectious disease with a worldwide distribution. Its causative agent, the intracellular bacterium Coxiella burnetii, infects a variety of vertebrate species, including humans. Its evolutionary origin remains almost entirely unknown and uncertainty persists regarding the identity and lifestyle of its ancestors. A few tick species were recently found to harbor maternally-inherited Coxiella-like organisms engaged in symbiotic interactions, but their relationships to the Q fever pathogen remain unclear. Here, we extensively sampled ticks, identifying new and atypical Coxiella strains from 40 of 58 examined species, and used this data to infer the evolutionary processes leading to the emergence of C. burnetii. Phylogenetic analyses of multi-locus typing and whole-genome sequencing data revealed that Coxiella-like organisms represent an ancient and monophyletic group allied to ticks. Remarkably, all known C. burnetii strains originate within this group and are the descendants of a Coxiella-like progenitor hosted by ticks. Using both colony-reared and field-collected gravid females, we further establish the presence of highly efficient maternal transmission of these Coxiella-like organisms in four examined tick species, a pattern coherent with an endosymbiotic lifestyle. Our laboratory culture assays also showed that these Coxiella-like organisms were not amenable to culture in the vertebrate cell environment, suggesting different metabolic requirements compared to C. burnetii. Altogether, this corpus of data demonstrates that C. burnetii recently evolved from an inherited symbiont of ticks which succeeded in infecting vertebrate cells, likely by the acquisition of novel virulence factors.