Detection of Brucella antibodies in domestic animals of southern Cameroon: Implications for the control of brucellosis.

Brucellosis is one of the world's most widespread bacterial zoonoses caused by Brucella. It leads to considerable economic losses as a result of low productivity of infected animals and the long debilitating illness in humans. Despite its impact on human and animal health, little attention has been paid on Brucella infections in domestic animals. It is in this light that the prevalence of Brucella antibodies was determined in domestic animals with the overarching goal of improving our knowledge on brucellosis in southern Cameroon. During cross-sectional studies conducted from December 2016 to August 2018 in five sites of southern Cameroon, blood samples were collected in cattle, sheep, goat, pig and dog. Plasma was obtained from each blood sample and Brucella antibodies were detected using the Rose Bengal test and the enzyme-linked immunosorbent assay (ELISA). From 1873 animals that were sampled, the overall prevalence of Brucella antibodies using Indirect enzyme-linked immunosorbent assay (i-ELISA) was 6.35% (118/1873): 9.12% (78/855) in cattle; 8.04% (30/373) in sheep; 6.06% (2/33) in dog, 1.87% (3/160) in pig and 1.1% (5/452) in goat. Between animal species (p-value < .0001, x2  = 33.63) as well as sampling sites (p-value = .0001, x2  = 18.97), significant differences were observed in the prevalence of Brucella antibodies. Yoko and Noun localities have shown the highest prevalence of 8.6% (30/348) and 7.2% (78/1070), respectively. This prevalence was significantly higher (p = .03, x2  = 1.25) in female than male cattle. Between adult (16.923%) and young cattle (7.8%), significant difference (p = .04, x2  = 6.42) was observed in the prevalence of Brucella antibodies. This study shows that the prevalence of Brucella antibodies varies between animal species and localities. It also shows several domestic animals of southern Cameroon that have been in contact with Brucella. It enabled to identify villages where investigations on the transmission dynamic must be focused for the final goal of developing control measures for this neglected zoonotic disease.

First detection of Theileria parva in cattle from Cameroon in the absence of the main tick vector Rhipicephalus appendiculatus.

A major risk factor for the spread of livestock diseases and their vectors is the uncontrolled transboundary movement of live animals for trade and grazing. Such movements constrain effective control of tick-transmitted pathogens, including Theileria parva. Only limited studies have been undertaken to identify ticks and tick-borne diseases (TTBDs) affecting cattle in central African countries, including Cameroon. We hereby report the collection of baseline data on the prevalence of T. parva in Cameroon through a countrywide cross-sectional survey, conducted in 2016, involving collection of blood samples from cattle from 63 sites across the five agro-ecological zones (AEZs) of the country. ELISA-based surveillance of infected cattle was performed on 479 randomly selected samples and revealed specific antibodies to T. parva in 22.7% and T. mutans in 41.1% of cattle. Screening of 1,340 representative DNA samples for the presence of T. parva identified 25 (1.86%) positives using a p104 antigen gene-based nested PCR assay. The positives were distributed across agro-ecological zones I, II, III and V. None of the p104 positive cattle exhibited clinical symptoms of East Coast fever (ECF). Using reverse line blot (RLB), 58 (4.3%) and 1,139 (85%) of the samples reacted with the T. parva and T. mutans oligonucleotide probes, respectively. This represents the first report of T. parva from Cameroon. Surprisingly, no Rhipicephalus appendiculatus ticks, the main vector of T. parva, were identified in a parallel study involving comprehensive morphological and molecular survey of tick species present in the country. Only two of the 25 p104 positive cattle were PCR-positive for the CD8+ T-cell target schizont-expressed antigen gene Tp1. Cloning and sequencing of Tp1 amplicons revealed sequence identity with the reference T. parva Muguga. This new finding raises serious concerns of a potential spread of ECF into the central African region.

A comprehensive survey of the prevalence and spatial distribution of ticks infesting cattle in different agro-ecological zones of Cameroon.

BACKGROUND: Ticks and tick-borne diseases are a major impediment to livestock production worldwide. Cattle trade and transnational transhumance create risks for the spread of ticks and tick-borne diseases and threaten cattle production in the absence of an effective tick control program. Few studies have been undertaken on cattle ticks in the Central African region; therefore, the need to assess the occurrence and the spatial distribution of tick vectors with the aim of establishing a baseline for monitoring future spread of tick borne-diseases in the region is urgent. RESULTS: A total of 7091 ixodid ticks were collected during a countrywide cross-sectional field survey and identified using morphological criteria. Of these, 4210 (59.4%) ticks were Amblyomma variegatum, 1112 (15.6%) Rhipicephalus (Boophilus) microplus, 708 (10.0%) Rhipicephalus (Boophilus) decoloratus, 28 (0.4%) Rhipicephalus (Boophilus) annulatus, 210 (3.0%) Hyalomma rufipes, 768 (10.8%) Hyalomma truncatum, and 19 (0.3%) Rhipicephalus sanguineus. Three ticks of the genus Hyalomma spp. and 33 of the genus Rhipicephalus spp. were not identified to the species level. Cytochrome c oxidase subunit 1 (cox1) gene sequencing supported the data from morphological examination and led to identification of three additional species, namely Hyalomma dromedarii, Rhipicephalus sulcatus and Rhipicephalus pusillus. The finding of the invasive tick species R. microplus in such large numbers and the apparent displacement of the indigenous R. decoloratus is highly significant since R. microplus is a highly efficient vector of Babesia bovis. CONCLUSIONS: This study reports the occurrence and current geographical distribution of important tick vectors associated with cattle in Cameroon. It appears that R. microplus is now well established and may be displacing native Rhipicephalus (Boophilus) species, such as R. decoloratus. This calls for an urgent response to safeguard the livestock sector in western central Africa.

A countrywide molecular survey leads to a seminal identification of the invasive cattle tick Rhipicephalus (Boophilus) microplus in Cameroon, a decade after it was reported in Cote d'Ivoire.

The cattle tick Rhipicephalus microplus is the most important arthropod vector of livestock diseases globally. Since its introduction in West Africa a decade ago, it has been reported in Ivory Coast, Benin, Togo, Mali, Burkina Faso and Nigeria with potentially far-reaching adverse impacts on the livestock sector in the region. Cameroon is located on a major route for transboundary cattle trade between Central and West Africa and it is therefore at risk from R. microplus invasion. This study investigated the occurrence of R. microplus in Cameroon, the genetic polymorphism of the tick and population structure of isolates from different regions of the country to provide data that underpin the design of future vector control programs. A cross-sectional survey was conducted in which ticks were collected from cattle at 54 sites across the five Agroecological zones (AEZs) within Cameroon. Tick identity (sex and species) was assigned using taxonomic keys. Species identity was confirmed through amplification and sequencing of the mitochondrial COI and 16S rRNA genes. A total of 7091 ticks were collected out of which 1112 (15.6%) were morphologically identified as R. microplus. The presence of R. microplus was confirmed in 4 out of 5 agroecological zones. Only two haplotypes were identified by both COI and 16S rRNA genes, indicating a very low divergence in the genetic structure of the R. microplus population in Cameroon. 16S rRNA sequence analysis revealed a new haplotype specific to Cameroon. Phylogenetic trees revealed that all isolates of R. microplus from Cameroon were grouped into the previously described Africa/Americas clade. Application of a niche modelling algorithm to R. microplus distribution in Cameroon predicted that suitable habitat for the tick extended into southern Nigeria. This study demonstrated for the first time the presence of R. microplus in Cameroon. Genetic diversity tests indicate that the tick has not evolved significantly since the initial introduction to West Africa. We suggest further longitudinal studies to better define the spatial and temporal expansion of the range of the tick and the drivers of this spread.

Identification of different trypanosome species in the mid-guts of tsetse flies of the Malanga (Kimpese) sleeping sickness focus of the Democratic Republic of Congo.

BACKGROUND: The Malanga sleeping sickness focus of the Democratic Republic of Congo has shown an epidemic evolution of disease during the last century. However, following case detection and treatment, the prevalence of the disease decreased considerably. No active survey has been undertaken in this focus for a couple of years. To understand the current epidemiological status of sleeping sickness as well as the animal African trypanosomiasis in the Malanga focus, we undertook the identification of tsetse blood meals as well as different trypanosome species in flies trapped in this focus. METHODS: Pyramidal traps were use to trap tsetse flies. All flies caught were identified and live flies were dissected and their mid-guts collected. Fly mid-gut was used for the molecular identification of the blood meal source, as well as for the presence of different trypanosome species. RESULTS: About 949 Glossina palpalis palpalis were trapped; 296 (31.2%) of which were dissected, 60 (20.3%) blood meals collected and 57 (19.3%) trypanosome infections identified. The infection rates were 13.4%, 5.1%, 3.5% and 0.4% for Trypanosoma congolense savannah type, Trypanosoma brucei s.l., Trypanosoma congolense forest type and Trypanosoma vivax, respectively. Three mixed infections including Trypanosoma brucei s.l. and Trypanosoma congolense savannah type, and one mixed infection of Trypanosoma vivax and Trypanosoma congolense savannah type were identified. Eleven Trypanosoma brucei gambiense infections were identified; indicating an active circulation of this trypanosome subspecies. Of all the identified blood meals, about 58.3% were identified as being taken on pigs, while 33.3% and 8.3% were from man and other mammals, respectively. CONCLUSION: The presence of Trypanosoma brucei in tsetse mid-guts associated with human blood meals is indicative of an active transmission of this parasite between tsetse and man. The considerable number of pig blood meals combined with the circulation of Trypanosoma brucei gambiense in this focus suggests a transmission cycle involving humans and domestic animals and could hamper eradication strategies. The various species of trypanosomes identified in the Malanga sleeping sickness focus indicates the coexistence of animal and human African Trypanosomiasis. The development of new strategies integrating control measures for human and animal trypanosomiasis may enable the reduction of the control costs in this locality.