Old Agents and Novel Variants of Tick-borne Microorganisms From Angola, 2017

The study of microorganisms from ticks collected in cattle from Angola is reported herein, demonstrating the circulation of the pathogen R. aeschlimannii and potential novel tick-borne microorganisms with unknown pathogenicity belonging to Ehrlichia, Spiroplasma, Coxiella, Babesia and Francisella spp. and corroborating the presence of Rickettsia africae and Babesia bigemina.

Molecular survey of cattle ticks in Burundi: First report on the presence of the invasive Rhipicephalus microplus tick

A recent research study on prevalence of tick-borne pathogens in Burundi reported high prevalence and endemicity of Theileria parva, Anaplasma marginale and Babesia bigemina infections in cattle. Detailed information about tick species infesting animals, their distribution and genetic diversity in Burundi is outdated and limited. This study therefore assessed the prevalence and genetic diversity of tick species infesting cattle across agroecological zones (AEZs) in Burundi. A cross-sectional study on the occurrence of tick species was conducted in 24 districts of Burundi between October and December 2017. Differential identification and characterization of ticks collected was conducted using tick morphological keys and molecular tools (cox1 and 12S rRNA gene). Chi-square test was used to test for association between agroecological zones and the prevalence of tick species. Phylogenetic relationships were inferred using bayesian and maximum likelihood algorithms. A total of 483 ticks were collected from the five AEZs sampled. Six tick species comprising of Rhipicephalus appendiculatus, R. sanguineus, R. evertsi evertsi, R. microplus, R. decoloratus and Amblyomma variegatum were observed. Rhipicephalus appendiculatus were the most prevalent ticks (~45%). A total of 138 specimens (28%) were found to be Rhipicephalus microplus, suggesting an emerging threat for cattle farmers. Twelve R. appendiculatus cox1 haplotypes were obtained from 106 specimens that were sequenced. Two cox1 haplotypes of R. microplus which clustered into previously reported Clade A were observed. Rhipicephalus sanguineus and R. evertsi evertsi ticks, the vectors of numerous zoonotic pathogens, were collected from cattle, which constitute a high risk for public health. These findings reveal an overlapping distribution of tick vectors in Burundi. The design of ticks and tick-borne diseases control strategies should consider the distribution of different vectors across the AEZs particularly the presence of the highly invasive R. microplus tick in Burundi and the potential risk of introducing the pathogenic Babesia bovis.

Detection of Babesia spp. in High Altitude Cattle in Ecuador, Possible Evidence of the Adaptation of Vectors and Diseases to New Climatic Conditions

Background: Babesia species are intraerythrocytic protozoa, distributed in tropical and subtropical areas of the world, causing anemic diseases in many animals, including cattle. This disease, called babesisosis, is transmitted from one animal to another through ticks (Tick Borne-Disease or TBD). On the other hand, Ecuador has a tropical climate that allows the development of the vector Rhipicephalus microplus, and therefore favors the transmission of Babesia spp. in cattle. Methods and principal findings: We determined the presence of Babesia spp. by PCR using 18s ribosomal gene as target (18s PCR) in 20 farms in the area of El Carmen (zone below 300 m above sea level) and 1 farm in Quito (2469 m.a.s.l.). In addition, we analyzed parameters such as age, sex, and packed cell volume (PCV) as explanatory variable associated with the disease. Results: The 18s PCR test showed that 18.94% (14.77% Babesia bovis and 4.17% Babesia bigemina) and 20.28% (14.69% B. bovis and 5.59% B. bigemina) of the cattle were positive for Babesia spp in farms sampled in El Carmen and in Quito, respectively. Age influenced the presence of animals positive for Babesia spp., but sex and PCV did not. The phylogenetic analysis of sequences showed 4 isolates of B. bovis and 3 isolates of B. bigemina in the 2 study zones, with similarities between 99.73 and 100% with other sequences. One B. bovis isolate was similar in the zone of El Carmen and Quito. Conclusion and significance: This work is the first molecular characterization of B. bigemina and B. bovis in Ecuador, and it is also the first evidence of Babesia spp. in cattle in the area of Quito at an altitude of 2469 m.a.s.l., being the highest altitude reported for animals with babesiosis and for the tick R. microplus. Climatic factors as well as mobility of tick-carrying animals without any control allow the presence of Babesiosis outbreaks in new geographical areas.

Molecular prevalence and risk factors associated with tick-borne pathogens in cattle in western Kenya

Background Tick-borne pathogens (TBPs) are of global importance, especially in sub-Saharan Africa where they represent a major constraint to livestock production. Their association with human disease is also increasingly recognized, signalling their zoonotic importance. It is therefore crucial to investigate TBPs prevalence in livestock populations and the factors associated with their presence. We set out to identify TBPs present in cattle and to determine associated risk factors in western Kenya, where smallholder livestock production is important for subsistence and market-driven income. Results Tick-borne pathogen infections in blood samples collected from cattle at livestock markets and slaughterhouses between May 2017 and January 2019 were identified by high-resolution melting analysis and sequencing of PCR products of genus-specific primers. Of the 422 cattle sampled, 30.1% (127/422) were infected with at least one TBP, while 8.8% (37/422) had dual infections. Anaplasma spp. (19.7%) were the most prevalent, followed by Theileria (12.3%), Ehrlichia (6.6%), and Babesia (0.2%) spp. Sequence analysis of the TBPs revealed them to be Anaplasma platys-like organisms (13.5%), Theileria velifera (7.4%), Anaplasma marginale (4.9%), Theileria mutans (3.1%), Theileria parva (1.6%), and Babesia bigemina (0.2%). Ehrlichia ruminantium, Rickettsia spp., and arboviruses were not detected. Exotic breeds of cattle were more likely to be infected with A. marginale compared to local breeds (OR: 7.99, 95% CI: 3.04–22.02, p < 0.001). Presence of ticks was a significant predictor for Anaplasma spp. (OR: 2.18, 95% CI: 1.32–3.69, p = 0.003) and Ehrlichia spp. (OR: 2.79, 95% CI: 1.22–7.23, p = 0.022) infection. Cattle sampled at slaughterhouses were more likely to be positive for Anaplasma spp. (OR: 1.64, 95% CI: 1.01–2.70, p = 0.048) and A. marginale (OR: 3.84, 95% CI: 1.43–12.21, p = 0.012), compared to those sampled at livestock markets. Conclusion This study reports TBP prevalence and associated risk factors in western Kenya, factors which are key to informing surveillance and control measures.

Frequência de Babesia spp. em bezerras holandesas no município de Patos de Minas - MG / Babesia spp. occurance in holstine calves in Patos de Minas - MG

As babesioses bovinas tratam-se de doenças parasitárias causadas por Babesia bovis e Babesia bigemina, sendo que o único vetor biológico é o carrapato Rhipicephalus (Boophilus) microplus. O aspecto mais relevante da patogenicidade desse agente reside na anemia grave, ocasionando um alto índice de morbimortalidade nos rebanhos, além de queda da produção de leite e carne, abortos, redução de fertilidade, entre outros prejuízos. A raça holandesa tem origem nos países baixos europeus e tem sido evidenciada como uma das principais raças leiteiras entre as bovinas, sendo expandida a criação da mesma em quase todos os continentes. Todas as raças bovinas são suscetíveis à Babesia spp., mas o Bos indicus apresenta maior resistência do que o Bos taurus. O Brasil é considerado como um país enzoótico para a Babesiose devido à constante transmissão dos agentes. O objetivo geral deste trabalho é determinar a prevalência de infecção por Babesia bovis em bezerras da raça Holandesa no município de Patos de Minas, MG, através da análise de esfregaços de sangue periférico. O estudo foi realizado em 10 (dez) propriedades de rebanho leiteiro da raça Holandesa (HPB), no período de julho de 2016. Foram avaliadas bezerras entre 90 e 270 dias, utilizando-se amostras de sangue periférico de 160 animais destas propriedades. Os resultados obtidos demonstram que (46%) dos animais foram considerados infectados por Babesia spp., confirmando a presença deste hemoparasito no município Patos de Minas, MG.

Molecular Detection of Tick-Borne Pathogens in American Bison (Bison bison) at El Uno Ecological Reserve, Janos, Chihuahua, Mexico

American bison (Bison bison) is listed as near-threatened and in danger of extinction in Mexico. Recent studies have demonstrated the presence of several emerging pathogens at the Janos Biosphere Reserve (JBR), inhabited by one wild herd of American bison. Blood samples were collected from 26 American bison in the JBR. We tested for the presence of Anaplasma marginale, Babesia bigemina, B. bovis, Borrelia burgdorferi sensu lato, and Rickettsia rickettsii DNA using nested and semi-nested PCR protocols performing duplicates in two different laboratories. Results showed three animals (11.5%) positive for B. burgdorferi s. l., three more (11.5%) for Rickettsia rickettsii, and four (19.2%) for B. bovis. Two individuals were co-infected with B. burgdorferi s. l. and B. bovis. We found no animals positive for A. marginale and B. bigemina. This is the first report in America of R. rickettsii in American bison. American bison has been described as an important reservoir for pathogens of zoonotic and veterinary importance; thus, the presence of tick-borne pathogen DNA in the JBR American bison indicates the importance of continuous wildlife health surveys.

Multiple vector-borne pathogens of domestic animals in Egypt

Vector Borne Diseases (VBDs) are considered emerging and re-emerging diseases that represent a global burden. The aim of this study was to explore and characterize vector-borne pathogens in different domestic animal hosts in Egypt. A total of 557 blood samples were collected from different animals using a convenience sampling strategy (203 dogs, 149 camels, 88 cattle, 26 buffaloes, 58 sheep and 33 goats). All samples were tested for multiple pathogens using quantitative PCR and standard PCR coupled with sequencing. We identified Theileria annulata and Babesia bigemina in cattle (15.9 and 1.1%, respectively), T. ovis in sheep and buffaloes (8.6 and 7.7%, respectively) and Ba. canis in dogs (0.5%) as well as Anaplasma marginale in cattle, sheep and camels (20.4, 3.4 and 0.7%, respectively) and Coxiella burnetii in sheep and goats (1.7 and 3%; respectively). New genotypes of An. centrale, An. ovis, An. platys-like and Borrelia theileri were found in cattle (1.1,3.4, 3.4 and 3.4%, respectively), An. platys-like in buffaloes (7.7%), An. marginale, An. ovis, An. platys-like and Bo. theileri in sheep (3.4, 1.7, 1.7 and 3.4%, respectively), An. platys, An. platys-like and Setaria digitata in camels (0.7, 5.4 and 0.7%, respectively) and Rickettsia africae-like, An. platys, Dirofilaria repens and Acanthocheilonema reconditum in dogs (1.5, 3.4, 1 and 0.5%, respectively). Co-infections were found in cattle, sheep and dogs (5.7, 1.7, 0.5%, respectively). For the first time, we have demonstrated the presence of several vector-borne zoonoses in the blood of domestic animals in Egypt. Dogs and ruminants seem to play a significant role in the epidemiological cycle of VBDs.

Development of Novel Multiplex PCR for Diagnosis of Co-infected Hemo-parasites in Cattle

Background: A novel, rapid and specific multiplex polymerase chain reaction was developed to diagnose hemo-parasitic infection in bovine blood co-infected with three of the most common hemo-parasites. Methods: The diagnostic process relied on the detection of the three different bovine hemoparasites isolated from red blood cells (RBCs) of cattle (N=30) by conventional Giemsa stained blood smear (GSBS) and confirmed by multiplex PCR. The multiplex PCR system was used to diagnose GSBS positive blood samples (N=12) found infected or co-infected with hemoparasites. The designed multiplex primer sets was attempted to amplify 205, 313 and 422 bp fragments of apocytochrome b, sporozoite and macroschizont 2 (spm2) and 16S rRNA gene for Babesia bigemina, Theileria annulata and Anaplasma marginale, respectively. Result: This multiplex PCR was sensitive with the ability to detect the presence of 150 ng of genomic DNA. The primers used in this multiplex PCR also showed highly specific amplification of specific gene fragments of each respective parasite. Comparing the two detection methods revealed that 58.33% of specimens showed concordant diagnoses with both techniques. The specificity, positive predictive value and kappa coefficient of the agreement was highest for diagnosis of B. bigemina and lowest for A. marginale. The overall Kappa coefficient for diagnosis based on GSBS for multiple pathogens compared to multiplex PCR was 0.56, slightly behind the threshold of 0.6 of agreement. Therefore, confirmation should always be based on PCR to rule out false positives due to differences in subjective observations, stain particles and false negatives due to low parasitemia. The simplicity and rapidity of this specific multiplex PCR method make it suitable for large-scale epidemiological studies and follow-up of drug treatments.