Background: Public interest for tick-borne pathogens in cattle livestock is rising due to their veterinary and zoonotic importance. Consequently, correct identification of these potential pathogens is crucial to estimate the level of exposition, the risk and the detrimental impact on livestock and the human population.Results: Conventional PCR with generic primers was used to identify groups of tick-borne pathogens in cattle breeds from northern Cameroon. The overall prevalence in 1260 blood samples was 89.1%, with 993 (78.8%) positive forTheileria/Babesiaspp., 959 (76.1%) forAnaplasma/Ehrlichiaspp., 225 (17.9%) forBorreliaspp., and 180 (14.3%) forRickettsia spp. Sanger sequencing of a subset of positively-tested samples revealed the presence ofTheileria mutans(92.2%, 130/141),T. velifera(16.3%, 23/141),Anaplasma centrale(10.9%, 15/137),A. marginale(30.7%, 42/137),A. platys(51.1%, 70/137),Anaplasmasp. ‘Hadesa’ (10.9%, 15/137),Ehrlichia ruminantium(0.7%, 1/137),E. canis(0.7%, 1/137),Borrelia theileri(91.3%, 42/46), Rickettsia africae(59.4%, 19/32) andR. felis(12.5%, 4/32). A high level of both intra- and inter-generic co-infections (76.0%) was observed. To the best of our knowledge,B. theileri, T. mutans,T. velifera,A. platys,Anaplasmasp. ‘Hadesa’,R. felisandE. canisare reported for the first time in cattle from Cameroon, and forR. felisit is the first discovery in the cattle host.Babesiaspp. were not detected by sequencing. The highest number of still identifiable species co-infections was up to four pathogens per genus group. Multifactorial analyses revealed a significant association of infection withBorrelia theileriand anemia. Whereas animals of older age had a higher risk of infection, the Gudali cattle had a lower risk compared to the other local breeds.Conclusion: Co-infections of tick-borne pathogens with an overall high prevalence were found in all five study sites, and were more likely to occur than single infections. Fulani, Namchi and Kapsiki were the most infected breed in general; however, with regions as significant risk factor. A better-adapted approach for tick-borne pathogen identification in co-infected samples is a requirement for epidemiological investigations and tailored control measures.

Background: African indigenous taurine cattle display unique adaptive traits shaped by husbandry management, regional climate and exposure to endemic pathogens. They are less productive with respect to milk and meat production which has been associated with amongst others, small size, traditional beliefs, husbandry practices, limited feed resources, disease burden and lack of sustained breeding for trait improvement. This resulted in the severe dwindling of their population size rendering them vulnerable to extinction.The Namchi taurine cattle breed is referred to as [Namchi (Doayo)] and shows resistance traits against trypanosome infection and exposure to tick infestation. Nonetheless, the historically later introduced Zebu cattle are the main cattle breeds in Africa today, even though they suffer more from locally prevailing pathogens.By using a whole genome sequencing approach, we sequenced with high depth for the first time the genomes of five cattle breeds from Cameroon in order to provide a valuable genetic resource for future African cattle breeding: the Namchi, an endangered trypano-tolerant taurine breed, the Kapsiki, an indigenous trypano-susceptible taurine breed, and three Zebu (Bos indicus indicus) breeds: Ngaoundere Gudali, White Fulani and Red Fulani.Results: Approximately 167 Gigabases of raw sequencing data were generated for each breed and mapped to the cattle reference genomes ARS-UCD1.2 and UMD3.1.The coverage was 103 to 140-fold when aligning the reads to ARS-UCD1.2 with an average mapping rate of ~ 99%, and 22 to 30-fold when aligning the reads to UMD3.1 with an average mapping rate of ~ 64%. The single nucleotide polymorphisms (SNPs) obtained from analysis using the genome ARS-UCD1.2 were compared with reference genomes of EuropeanBos taurusHolstein, the AsianBos indicusBrahman, and the African trypanotolerant N’Dama breeds.A total of ~ 100 million (M) SNPs were identified and 7.7 M of those were breed-specific. An approximately 11.1 M constituted of small insertions and deletions. By using only breed-specific non-synonymous variants we identified genes as genetic signatures and associated Gene Ontology (GO) terms that could explain certain cattle-breed specific phenotypes such as increased tolerance against trypanosome parasites in the Namchi breed and heat tolerance in the Kapsiki breed. Phylogenetic analysis grouped, except for Namchi, theBos taurusbreeds Kapsiki, N’Dama and Holstein together while theB. indicusbreeds White and Red Fulani, Gudali and Brahman clustered separately. The deviating result for Namchi indicates a hybrid status of the selected animal with a recent introgression of Zebu genes into its genome.Conclusions: The findings provide the first comprehensive set of genome-wide variant data of the most important Cameroonian cattle breeds. The genomic data shall constitute a foundation for breed amelioration whilst exploiting the heritable traits and support conservation efforts for the endangered local cattle breeds.

Background: African animal trypanosomosis remains the major constraint of livestock production and livelihood of pastoral communities in Cameroon. Despite several decades of vector and parasite control efforts, it has not been eradicated. Alternative and sustainable control strategies require a sound knowledge of the local species, strains and vectors. In the Sudano-Sahelian and Guinea Savannah of Cameroon the prevalence and genetic diversity of trypanosomes infecting cattle was investigated by microscopy of cattle blood buffy coat and molecular methods using generic primers targeting parts of the internal transcribed spacer 1 (ITS-1) and encoded glycosomal glyceraldehyde 3-phosphate dehydrogenase-gene (gGAPDH). Results: A total of 1176 randomly chosen cattle from five divisions in the Sudano-Sahelian and Guinea Savannah of Cameroon were examined. The overall prevalence of trypanosomes by microscopy was 5.9% (56/953) in contrast to 53.2% (626/1176) when molecular tools were used. This indicated a limited sensitivity of microscopy in subclinical infections with frequently low parasitemia. Three trypanosome species were identified by light microscopy: T. vivax (2.3%), T. brucei (3.7%) and T. congolense (3.0%), whereas five were identified by PCR, namely T. grayi/T. theileri (30.8%), T. vivax (17.7%), T. brucei (14.5%) and T. congolense (5.1%). Unexpected cases of T. grayi (n = 4) and T. theileri (n = 26) were confirmed by sequencing. Phylogenetic analysis of the gGAPDH revealed the presence of T. vivax, clade A and T. vivax clade C, which were co-endemic in the Faro et Deo division. T. grayi/T. theileri were the predominant species infecting cattle in tsetse free areas. In contrast, T. vivax, T. brucei and T. congolense were more abundant in areas where the Glossina-vectors were present. Conclusions: The abundance of pathogenic trypanosomes in tsetse infested areas is alarming and even more, the occurrence of T. vivax, T. brucei, T. congolense, T. theileri and T. grayi in tsetse-free areas implies that tsetse control alone is not sufficient to control trypanosomosis in livestock. To implement control measures that reduce the risk of spread in tsetse free areas, close monitoring using molecular tools and a thorough search for alternative vectors of trypanosomes is recommended.

Ticks are obligate blood-sucking parasites of wild animals and transmit many zoonotic microorganisms that can spread to domesticated animals and then to humans. In Cameroon, little is known about tick diversity among wildlife, especially for animals which are hunted for human consumption. Therefore, this survey was undertaken to investigate tick and Rickettsia species diversity parasitizing the wild animals sold in bush meat markets in Cameroon. In total, 686 ticks were collected and identified to the species level based on morphology, and some were genetically analyzed using the 16S rRNA gene. Eighteen tick species belonging to five genera were identified: Amblyomma spp. (Amblyomma compressum, Amblyomma flavomaculatum, and Amblyomma variegatum), Haemaphysalis spp. (Haemaphysalis camicasi, Haemaphysalis houyi, Haemaphysalis leachi, and Haemaphysalis parmata), Hyalomma spp. (Hyalomma nitidum, Hyalomma rufipes, and Hyalomma truncatum), Ixodes spp. (Ixodes rasus and Ixodes moreli), and Rhipicephalus spp. (Rhipicephalus guilhoni, Rhipicephalus moucheti, Rhipicephalus muhsamae, Rhipicephalus microplus, Rhipicephalus camicasi, and Rhipicephalus linnaei). In terms of Rickettsia important for public health, two Rickettsia spp., namely Rickettsia aeschlimannii and Rickettsia africae, were detected in Hyalomma spp. and Amblyomma spp., respectively. Distinct tick–pathogen patterns were present for divergent sequences of R. africae associated with exclusively A. variegatum vectors (type strain) versus vectors comprising A. compressum, A. flavomaculatum, and A. variegatum. This suggests possible effects of vector species population dynamics on pathogen population circulation dynamics. Furthermore, Candidatus Rickettsia africaustralis was detected for the first time in Cameroon in I. rasus. This study highlights the high diversity of ticks among wildlife sold in bush meat markets in Cameroon.

The front line molecules from filarial worms and other nematodes or helminthes are their Excretory-Secretory (ES) products. Their interaction with the host cells, proteins and immune system accounts for the skin and eye pathology or hyposensitivity observed in human onchocerciasis. ES products and adult worms’ crude extracts from Onchocerca ochengi, a filarial nematode that infects the African zebu cattle, were utilized in the present study as a model for studying Onchocerca volvulus that causes river blindness in man. The ES products were generated from adult male and female worms in vitro and analyzed with poly acrylamide gel electrophoresis (PAGE) and enzyme-linked immunosorbent assay (ELISA) using sera from Onchocerca-infected cattle and humans. The cattle sera were collected from a herd that had been exposed for six years to natural transmission of Onchocerca spp. The expressed reactivity was evaluated and differences analyzed statistically using Kruskal-Wallis rank and Chi-square tests. The gel electrophoretic analyses of 156 ES products from O. ochengi female and male worms and of two somatic extracts from three females and 25 males revealed differences in the protein pattern showing pronounced bands at 15, 30–50 and 75 kDa for male ES proteins and 15, 25 and 40–75 kDa for somatic extracts, respectively and less than 100 kDa for female worms. Proteins in the ES products and somatic extracts from female and male Onchocerca ochengi worms were recognized by IgG in sera from both Onchocerca-exposed cattle and humans. Bovine serum antibodies reacted more strongly with proteins in the somatic extracts than with those in the ES products. Interestingly, the reaction was higher with male ES products than with ES products from female worms, suggesting that the males which migrate from one nodule to another are more exposed to the host immune system than the females which remain encapsulated in intradermal nodules. This study demonstrates that O. ochengi ES products and, in particular, extracts from male filariae may represent a good source of immunogenic proteins and potential vaccine candidates.

In onchocerciasis-endemic communities, black fly vectors of Onchocerca volvulus cause annoyance. No vector control was performed in Cameroon to complement ivermectin treatment despite high fly densities in the savannah and forest areas. This study assessed the knowledge and practices of four onchocerciasis-endemic communities in Sudan and the Guinea savannah in Cameroon. We surveyed four communities using a structured questionnaire. We interviewed 452 individuals (Mawong: 136, Befang: 160, Soramboum: 88, and Galim: 68) aged 14–50 years. Most respondents (90%) knew about black flies being a nuisance but had misconceptions about their bite’s effect, location, and season. Only 9.3% knew that black fly bites could lead to blindness, whereas 34.1% correctly regarded evenings as the biting period. In Savannah, Sudan, 30.9% of the interviewees stated that black flies bite both outdoors and indoors, and 37.0% of the enrolees believed that black flies bite any exposed body part. In the Guinea savannah, 61.1% of respondents agreed that black flies typically bite during the dry season. Proper clothing can protect against black fly bites, but the transmission mode and vector biology are poorly understood. Implementing community-based vector surveillance can help fight onchocerciasis and reduce black fly nuisance.