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 review of recent research on Theileria parva: Implications for the infection and treatment vaccination method for control of East Coast fever

The infection and treatment (ITM) live vaccination method for control of Theileria parva infection in cattle is increasingly being adopted, particularly in Maasai pastoralist systems. Several studies indicate positive impacts on human livelihoods. Importantly, the first detailed protocol for live vaccine production at scale has recently been published. However, quality control and delivery issues constrain vaccination sustainability and deployment. There is evidence that the distribution of T. parva is spreading from endemic areas in East Africa, North into Southern Sudan and West into Cameroon, probably as a result of anthropogenic movement of cattle. It has also recently been demonstrated that in Kenya, T. parva derived from cape buffalo can ‘breakthrough’ the immunity induced by ITM. However, in Tanzania, breakthrough has not been reported in areas where cattle co‐graze with buffalo. It has been confirmed that buffalo in northern Uganda national parks are not infected with T. parva and R. appendiculatus appears to be absent, raising issues regarding vector distribution. Recently, there have been multiple field population genetic studies using variable number tandem repeat (VNTR) sequences and sequencing of antigen genes encoding targets of CD8+ T‐cell responses. The VNTR markers generally reveal high levels of diversity. The antigen gene sequences present within the trivalent Muguga cocktail are relatively conserved among cattle transmissible T. parva populations. By contrast, greater genetic diversity is present in antigen genes from T. parva of buffalo origin. There is also evidence from several studies for transmission of components of stocks present within the Muguga cocktail, into field ticks and cattle following induction of a carrier state by immunization. In the short term, this may increase live vaccine effectiveness, through a more homogeneous challenge, but the long‐term consequences are unknown.     

Antigen gene and variable number tandem repeat (VNTR) diversity in Theileria parva parasites from Ankole cattle in south‐western Uganda: Evidence for conservation in antigen gene sequences combined with extensive polymorphism at VNTR loci

Theileria parva is a tick‐transmitted apicomplexan protozoan parasite that infects lymphocytes of cattle and African Cape buffalo (Syncerus caffer), causing a frequently fatal disease of cattle in eastern, central and southern Africa. A live vaccination procedure, known as infection and treatment method (ITM), the most frequently used version of which comprises the Muguga, Serengeti‐transformed and Kiambu 5 stocks of T. parva, delivered as a trivalent cocktail, is generally effective. However, it does not always induce 100% protection against heterologous parasite challenge. Knowledge of the genetic diversity of T. parva in target cattle populations is therefore important prior to extensive vaccine deployment. This study investigated the extent of genetic diversity within T. parva field isolates derived from Ankole (Bos taurus) cattle in south‐western Uganda using 14 variable number tandem repeat (VNTR) satellite loci and the sequences of two antigen‐encoding genes that are targets of CD8+T‐cell responses induced by ITM, designated Tp1 and Tp2. The findings revealed a T. parva prevalence of 51% confirming endemicity of the parasite in south‐western Uganda. Cattle‐derived T. parva VNTR genotypes revealed a high degree of polymorphism. However, all of the T. parva Tp1 and Tp2 alleles identified in this study have been reported previously, indicating that they are widespread geographically in East Africa and highly conserved.     

Monitoring vaccinated cattle for induction and longevity of persistent tick‐transmissible infection: Implications for wider deployment of live vaccination against East Coast fever in Tanzania

The infection and treatment (ITM) procedure remains the only available method of immunization against Theileria parva infection. One constraint to deployment is the perception that the carrier state induced by ITM could result in enhanced disease problems. More than one million cattle have been ITM vaccinated in pastoralist systems in Tanzania over the last 2 decades. We present the results of a longitudinal study of six groups of cattle in Maasai villages in northern Tanzania exposed to natural tick challenge for between 2 weeks and 14 years post‐vaccination. The p104 nested PCR revealed a higher frequency of T. parva carriers among vaccinates (30%) compared with controls (8%) (OR = 4.89, p = .000), with the highest frequency of carriers found in calves vaccinated 6 months previously, although carrier state was also detected in cattle vaccinated >10 years prior to the study. Variable number tandem repeat genotype analysis revealed 6 MS7 alleles with sizes ranging from 150 bp to 500 bp, but only two alleles were detected in cattle vaccinated >4 years earlier, relative to five alleles detected in recently vaccinated cattle and controls. In terms of heterozygosity, diversity was maximal in calves vaccinated within the last 2 weeks (h = 0.776) but lowest in cattle vaccinated 4 years earlier (h = 0.375). The analysis suggested close genetic relatedness of parasites in vaccinated and unvaccinated groups and up to 96% of variation was within rather than between the groups. These results confirm that ITM leads to a long‐term T. parva carrier state in cattle and the detection of vaccine component VNTR in co‐grazing unvaccinated cattle suggests potential vaccine transmission by ticks. However, vaccination stocks did not totally replace local genotypes, at least in cattle populations. These findings should mitigate concerns that ITM modifies T. parva field populations in a way that enhances disease in the medium term.     

Multilocus genotyping of Theileria parva isolates associated with a live vaccination trial in Kenya provides evidence for transmission of immunizing parasites into local tick and cattle populations

The live infection and treatment (ITM) vaccination procedure using the trivalent Muguga cocktail is increasingly being used to control East Coast fever, with potential implications for Theileria parva population genetic structure in the field. Transmission of the Kiambu V T. parva component to unvaccinated cattle has previously been described in Uganda. We monitored the T. parva carrier state in vaccinated and control animals on a farm in West Kenya where an ITM stabilate derived from the Kenyan T. parva Marikebuni stock was evaluated for field efficacy. A nested PCR‐based Marikebuni‐specific marker identified a carrier state in nine of ten vaccinated animals, detectable for a period of two years. We used 22 variable number tandem repeat (VNTR) markers to determine multilocus genotypes (MLGs) of 19 T. parva schizont‐infected lymphocyte isolates derived from cattle and field ticks. Two isolates from unimmunized cattle were identical to the Marikebuni vaccination stock. Two cattle isolates were identical to a Muguga cocktail component Kiambu V. Seven isolates from ticks exhibited MLGs that were identical to the Serengeti/Muguga vaccine stocks. Six cattle and two tick‐derived stocks exhibited unique MLGs. The data strongly suggest transmission of immunizing genotypes, from Marikebuni vaccine‐induced carrier cattle to unimmunized cattle. It is possible that genotypes similar to those in the Muguga cocktail are present in the field in Western Kenya. An alternative hypothesis is that these parasites may have originated from vaccine trial sites in Eastern Uganda. If correct, this suggests that T. parva stocks used for immunization can potentially be disseminated 125 km beyond the immediate vaccination site. Regardless of their origin, the data provide evidence that genotypes similar to those in the Muguga cocktail are circulating in the field in East Africa, alleviating concerns about dissemination of ‘alien’ T. parva germplasm through live vaccination.     

Molecular Demonstration of Trypanosoma evansi and Trypanosoma lewisi DNA in Wild Rodents from Cambodia,Lao PDR and Thailand

In this study, we investigated the molecular evidence of Trypanosoma evansi in wild rodents from Cambodia, Lao PDR and Thailand. Between November 2007 and June 2009, 1664 rodents were trapped at eight sites representative of various ecological habitats. Of those animals, 94 were tested by direct microscopic blood examination, 633 using the Card Agglutination Test for Trypanosomes (CATT/T. evansi) and 145 by Polymerase Chain Reaction (PCR) with two sets of primers: TRYP1 (amplifying ITS1 of ribosomal DNA of all trypanosomes) and TBR (amplifying satellite genomic DNA of Trypanozoon parasites). Using TRYP1, based on the size of the PCR products, 15 samples from the three countries were positive for Trypanosoma lewisi (two were confirmed by sequencing), and three were positive for Trypanozoon (one was confirmed by sequencing and three by TBR primers); the specificity of the primers failed as rodent DNA was amplified in some cases. Using TBR, six samples were positive for Trypanozoon (one was confirmed by sequencing); as T. evansi is the only species of the Trypanozoon sub‐genus possibly present in Asian rodents, these results confirmed its presence in rodents from Thailand (Rattus tanezumi) and Cambodia (R. tanezumi, Niviventer fulvescens & Maxomys surifer). Further investigations are necessary to establish the situation in Lao PDR. None of the 16 samples most strongly positive to the CATT proved to be positive for Trypanozoon by PCR. The merits of the CATT for such studies were not confirmed. Studying the urban and rural circulation of these parasites in rodents will enable an evaluation of human exposure and infection risk, as human infections by T. evansi were recently described in India and by T. lewisi in India and Thailand. As sequencing PCR products is expensive, the development of new molecular and serological tools for rodents would be very useful.     

Molecular detection and genetic diversity of Bartonella species in large ruminants and associated ectoparasites from the Brazilian Cerrado

Currently, five Bartonella species and an expanding number of Candidatus Bartonella species have globally been reported in ruminants. Likewise, different Bartonella genotypes were identified. However, studies relating to ruminant‐associated Bartonella in Brazil are scarce. The current study aimed to assess the prevalence and genetic diversity of Bartonella in cattle, buffaloes and associated ectoparasites in Brazil. For this purpose, EDTA‐blood samples from 75 cattle and 101 buffaloes were sampled. Additionally, 128 Rhipicephalus microplus and one Amblyomma sculptum ticks collected from cattle, and 197 R. microplus, one A. sculptum and 170 lice (Haematopinus tuberculatus) collected from buffaloes were included. Bartonella DNA was initially screened through an HRM real‐time PCR assay targeting the 16S–23S internal transcribed spacer (ITS), and the positive samples were submitted to an additional HRM assay targeting the ssrA gene. The HRM‐positive amplicons were sequenced, and the nucleotide identity was assessed by BLASTn. Bartonella spp.‐positive DNA samples were analysed by conventional PCR assays targeting the gltA and rpoB genes, and then, the samples were cloned. Finally, the phylogenetic positioning and the genetic diversity of clones were assessed. Overall, 21 of 75 (28%) cattle blood samples and 13 of 126 (10.3%) associated ticks were positive for Bartonella bovis. Out of 101 buffaloes, 95 lice and 188 tick DNA samples, one (1%) buffalo and four (4.2%) lice were positive for Bartonella spp. Conversely, none of the ticks obtained from buffaloes were positive for Bartonella. The Bartonella sequences from buffaloes showed identity ranging from 100% (ITS and gltA) to 94% (ssrA) with B. bovis. In contrast, the Bartonella DNA sequences from lice were identical (100%) to uncultured Bartonella sp. detected in cattle tail louse (Haematopinus quadripertusus) from Israel in all amplified genes. The present study demonstrates the prevalence of new B. bovis genotypes and a cattle lice‐associated Bartonella species in large ruminants and their ectoparasites from Brazil. These findings shed light on the distribution and genetic diversity of ruminant‐ and ectoparasite‐related Bartonella in Brazil.     

Serum‐free in vitro cultivation of Theileria annulata and Theileria parva schizont‐infected lymphocytes

Theileriosis is a tick‐borne disease caused by intracellular protozoa of the genus Theileria. The most important species in cattle are Theileria annulata and Theileria parva. Both species transform leucocyte host cells, resulting in their uncontrolled proliferation and immortalization. Vaccination with attenuated T. annulata‐infected cell lines is currently the only practical means of inducing immunity in cattle. Culture media for Theileria spp. typically contain 10%–20% foetal bovine serum (FBS). The use of FBS is associated with several disadvantages, such as batch‐to‐batch variation, safety and ethical concerns. In this study, the suitability of serum‐free media for the cultivation of Theileria‐transformed cell lines was examined. Three commercial serum‐free media (HL‐1, ISF‐1 and Hybridomed DIF 1000) were evaluated for their ability to support growth of the T. annulata A288 cell line. The generation doubling times were recorded for each medium and compared with those obtained with conventional FBS‐containing RPMI‐1640 medium. ISF‐1 gave the shortest generation doubling time, averaging 35.4 ± 2.8 hr, significantly shorter than the 52.2 ± 14.9 hr recorded for the conventional medium (p = .0011). ISF‐1 was subsequently tested with additional T. annulata strains. The doubling time of a Moroccan strain was significantly increased (65.4 ± 15.9 hr) compared with the control (47.7 ± 7.5 hr, p = .0004), whereas an Egyptian strain grew significantly faster in ISF‐1 medium (43.4 ± 6.5 hr vs. 89.3 ± 24.8 hr, p = .0001). The latter strain also showed an improved generation doubling time of 73.7 ± 21.9 hr in an animal origin‐free, serum‐free, protein‐free medium (PFHM II) compared with the control. Out of four South African T. parva strains and a Theileria strain isolated from roan antelope (Hippotragus equinus), only one T. parva strain could be propagated in ISF‐1 medium. The use of serum‐free medium may thus be suitable for some Theileria cell cultures and needs to be evaluated on a case‐by‐case basis. The relevance of Theileria cultivation in serum‐free media for applications such as vaccine development requires further examination.     

Swab cloths as a tool for revealing environmental contamination by Q fever in ruminant farms

Q fever is a zoonotic abortive disease of ruminants mostly transmitted by inhalation of aerosols contaminated by Coxiella burnetii. Clusters of cases or even epidemics regularly occur in humans but, to date, there is no consensus about the best way to carry out outbreak investigations in order to identify potential farms at risk. Although environmental samples might be useful during such investigations, there are few baseline data on the presence of C. burnetii in the environment of ruminant farms. We thus investigated dust samples from cattle, sheep and goat farm buildings in order to (a) estimate C. burnetii detection frequency and bacterial loads in the environment, and (b) determine whether this environmental contamination is associated with series of abortions attributed to Q fever. We considered 113 herds with a recent abortive episode potentially related (n = 60) or not (n = 53) to C. burnetii. Dust was sampled using a swab cloth and tested by a quantitative PCR method targeting the IS1111 gene. Coxiella burnetii DNA was detected on 9 of 50 cattle farms, 13 of 19 goat farms and 30 of 40 sheep farms. On 16 cloths, bacterial loads were higher than 10⁸ genome equivalents, levels as high as in infectious materials such as placentas and aborted foetuses. Overall, the probability of detecting C. burnetii DNA was higher on small ruminant farms than cattle farms, in herds suspected of Q fever and in large herds. We conclude that swab cloths are a putative indicator of contamination of ruminant farms by C. burnetii.     

First detection and molecular identification of Neospora caninum from naturally infected cattle and sheep in North Africa

Neosporosis, caused by the protozoan Neospora caninum , is a major cause of reproductive failure in ruminants causing enormous economic losses. The objective of this study was to estimate the infection rate and molecular identification of N. caninum in Tunisian cattle and sheep. A total number of 348 meat samples were collected from 150 cows and 198 ewes slaughtered in the regional slaughterhouse of Béja (North‐west Tunisia) and tested for the presence of N. caninum ITS 1 gene using PCR followed by sequencing of some PCR products. A phylogenetic tree was then constructed to compare the partial sequences of the ITS 1 gene with GenBank sequences. The overall molecular infection prevalence of N. caninum was significantly higher in cattle than in sheep (22 and 10.6%, respectively, p  = .003). In sheep, the highest prevalence was observed in the northern Béja locality (31.2 ± 16.1), with the Noire de Thibar breed as the most infected sheep breed (31.7 ± 14.2%) (p  < .001). In cattle, there were no differences in the molecular prevalence of N. caninum according to breeds and localities. The association between age and N. caninum molecular prevalence was statistically significant in both species; the highest prevalence was observed in sheep of more than one year of age (19.4 ± 9.1%), and in cattle between two and eight years of age (28.8 ± 10.9%). Comparison of the partial sequences of the ITS 1 gene revealed 96%–100% similarity among our N. caninum amplicon and those deposited in GenBank. To our knowledge, this is the first detection and molecular identification of N. caninum in sheep and cattle in North Africa. This information is pertinent in designing control programmes that would reduce economic losses in the livestock industry.     

Q Fever Dairy Herd Status Determination Based on Serological and Molecular Analysis of Bulk Tank Milk

Ruminants are recognized as the main reservoirs of Coxiella burnetii. EFSA highlighted the lack of knowledge about Q fever prevalence in many European countries. A cross‐sectional study was carried out in randomly selected dairy herds (n = 109) from central Portugal to screen for C. burnetii infection and to correlate it with herd factors. Bulk tank milk (BTM) samples from cattle (n = 45) and small ruminant (n = 64) herds were tested by ELISA and PCR. The apparent seroprevalence of Q fever was estimated in 45.9% (95% CI: 36.3–55.7) being higher in small ruminants (51.6; 95% CI: 39.6–63.4) than in cattle (37.8; 95% CI: 25.1–52.4). The shedding of C. burnetii in BTM was detected in 11.9% (95% CI: 7.1–19.4) of BTM, and it was higher in cattle (20%; 95% CI: 10.9–33.8) than in sheep and mixed herds (6.3%; 95% CI: 2.5–15). A high bacterial load (≥ 3 × 10³ bacteria/ml) was observed in 85% of PCR‐positive BTM. A significant correlation was found between the bacterial load and positive samples on ELISA (P < 0.001). Antibody positivity was significantly associated with the increased herd size (P < 0.01) and the occurrence of abortion (P < 0.05), whereas the shedding of C. burnetii was significantly associated with the report of infertility (P < 0.05). The results highlight that serological and molecular methods in combination are a useful tool to screen for Q fever and to clarify the herd infection status. The shedding of C. burnetii through milk is important, especially in dairy cattle, and thus, the role of milk as a potential source of infection among dairy workers should not be neglected. To our knowledge, this is the first study reporting C. burnetii infection in dairy livestock in Portugal showing that Q fever is significant in dairy herds, leading to economic losses and being a risk for public health, which highlights the need of implementation of control measures.     

Phylogenetic profiling for zoonotic Ehrlichia spp. from ixodid ticks in the Eastern Cape,South Africa

Ticks are obligate hematophagous parasite of vertebrate that transmit a range of pathogenic microorganisms that can cause diseases in livestock and humans. The range of tick‐borne disease causative agents infecting domestic animals and humans has recently increased. Several significant zoonotic tick‐borne diseases such as ehrlichiosis among others are on the increase worldwide. This study was designed to investigate the occurrence of zoonotic Ehrlichia spp. from samples collected from livestock in selected communities in the Eastern Cape Province, South Africa. Tick samples were manually collected from domesticated animals in selected homesteads. The ticks were morphologically identified to species and tested for Ehrlichia infection via polymerase chain reaction (PCR), using genus‐specific disulphide bond formation protein (dsbA) gene primers. This was followed by sequence analysis of amplicons and phylogeny. Of the 1,200 ticks collected, Amblyomma hebraeum was most prevalent (n = 335; 27.9%), followed by Rhipicephalus appendiculatus (n = 274; 22.8%), Rhipicephalus decoloratus; (n = 224; 18.7%) and Rhipicephalus eversti eversti (n = 200, 16.7%). Ehrlichia DNA was detected in 19/1,200 (1.6%) of the screened DNA samples. A homology search of the generated sequences revealed a high percentage of identity between 95% and 98% with other homologous dsbA gene sequences of other Ehrlichia species in GenBank. Phylogenetic analysis showed that the obtained sequences clustered unambiguously with other Ehrlichia sequences from different geographical regions of the world. We concluded that Ehrlichial pathogens are vectored by the ticks collected from domesticated animals in the study areas, thus suggesting concern for public health, as some of the recovered pathogens are zoonotic in nature and could pose serious public health risk through human exposure to tick bites.     

First Molecular Identification and Genetic Characterization of Theileria lestoquardi in Sheep of the Maghreb Region

Theileria lestoquardi is the most prominent Theileria species in small ruminants that causes malignant theileriosis of sheep in Africa and Asia. In the present survey, blood samples and ticks were collected in Kebili (southern Tunisia) from 166 Queue Fine de l'Ouest sheep. Giemsa‐stained blood smears, immunofluorescent antibody test (IFAT) and PCR were performed. The DNA was extracted from blood and analysed by PCR targeting 18S rRNA gene of Theileria spp. and then sequenced. A total number of 140 ticks were collected from a total number of 166 sheep during the four seasons. The ticks belonged to two genera and 4 species; the most frequent tick was Hyalomma excavatum 84.3% (118/140) and then Rhipicephalus spp. 15.7% (22/140). Only two animals had positive Giemsa‐stained blood smears, and they were also positive by IFAT. The amplicons had 99.3 and 99.6% homology with the BLAST published T. lestoquardi amplicons. To our knowledge, this is the first report of T. lestoquardi in small ruminants within the Maghreb region.     

The first report of animal genotypes of Cryptosporidium parvum in immunosuppressed and immunocompetent humans in Slovakia

The aim of our study was to determine species and genotypes of Cryptosporidium in patients suffering from immunosuppressive illnesses, but also in immunocompetent patients suffering from diarrhoea. A total of 80 samples of faeces were collected from both immunosuppressed and immunocompetent patients. The immunosuppressed patients (65 samples) — 35 adult patients (group A) and 30 children (group B) were hospitalized at the Clinic of Oncohemathology. Samples from immunocompetent humans (15 samples, group C) were taken from patients with clinical signs of acute diarrhoea. With the use of molecular methods targeting the 60 kDa glycoprotein (GP60) gene region, we have identified multiple genotypes of Cryptosporidium. parvum and Cryptosporidium. hominis in immunocompromised, but also in immunocompetent individuals (C. hominis IbA10G2, IeA12G3T3; C. parvum IIaA10G1R1, IIaA11G2R1, IIaA12G2R1, IIaA13G1R1, IIaA14G1R1, IIaA14G2R1, IIaA17G1R1 and IIaA18G1R1). This is the first report of the occurrence of genotypes IIaA10G1R1, IIa12G2R1 and IIaA18G1R1 in human hosts.     

Molecular investigation of “Candidatus Mycoplasma haemobos” in goats and sheep in central China

Hemoplasma “Candidatus Mycoplasma haemobos” infections in cattle have been reported in East Asia, Europe, and South America, whereas same cases were documented in buffalo and cattle in Southern China. From April 2018 to May 2018, a mycoplasma epidemic was reported in the mountainous area of central China; Boophilus microplus has also been documented, causing severe haematuria in goats and sheep. The infected animals slowly recovered after diminazene aceturate and praziquantel treatment. To determine whether the hemoplasma caused this infection, 67 blood samples (42 from goats, 25 from sheep) and 132 B. microplus samples were collected for PCR and sequence analysis. The results showed that 19 out of the 42 goat blood samples, 10 out of the 25 sheep blood samples, and 70 out of the 132 B. microplus samples (53%) tested positive for “C. M. haemobos”. This study provides molecular evidence of “C. M. haemobos” infections in goat and sheep, and that B. microplus harbours “C. M. haemobos”.     

Rickettsia parkeri in free‐ranging wild canids from Brazilian Pampa

Spotted fevers are tick‐borne diseases associated with various Rickettsia species. Rickettsia parkeri sensu stricto (s.s.) is the agent of an emerging eschar‐associated rickettsiosis in humans from the USA and South American Pampa. Considering that R. parkeri s.s. is restricted to Americas and the potential role of dogs in the epidemiology of the disease, it is thus reasonable to hypothesize that wild canids could be involved in the enzootic cycle of this rickettsiosis. The aim of this work was to investigate the potential role of the wild canids from Pampa, Cerdocyon thous (crab‐eating fox) and Lycalopex gymnocercus (Pampas fox), in the ecology of R. parkeri s.s. For that, 32 live‐trapped free‐ranging wild canids were sampled. Ticks were observed in 30 of the 32 foxes. Of the 292 ticks collected, 22 (7.5%) were positive by PCR for the presence of R. parkeri s.s. DNA . Also, 20 (62%) wild canids showed antibodies against R. parkeri . The results suggest that wild canids are involved in the enzootic cycle of R. parkeri s.s. in the Pampa biome and could be responsible for pathogen (and its vectors) dispersal.     

Genetic diversity of Bartonella spp. in vampire bats from Brazil

Recently, an increasing number of Bartonella species have been emerged to cause human diseases. Among animal reservoirs for Bartonella spp., bats stand out due to their high mobility, wide distribution, social behaviour and long‐life span. Although studies on the role of vampire bats in the epidemiology of rabies have been extensively investigated in Latin America, information on the circulation and genetic diversity of Bartonella species in these bat species is scarce. In the present work, 208 vampire bats, namely Desmodus rotundus (the common vampire bat; n = 167), Diphylla ecaudata (the hairy‐legged vampire bat; n = 32) and Diaemus youngii (the white‐winged vampire bat; n = 9) from 15 different states in Brazil were sampled. DNA was extracted from liver tissue samples and submitted to real‐time PCR (qPCR) and conventional PCR (cPCR) assays for Bartonella spp. targeting five genetic loci, followed by phylogenetic and genotype network analyses. Fifty‐one out of 208 liver samples (24.51%) were positive for Bartonella DNA in the ITS real‐time PCR assay [40 (78.43%) of them were from D. rotundus from 11 states, and 11 (21.57%) samples from D. ecaudata from three states. Eleven genotypes were found for each gltA and rpoB genes. Several ITS sequences detected in the present study clustered within the lineage that includes B. bacilliformis and B. ancachensis. The Bayesian phylogenetic inference based on the gltA gene positioned the obtained sequences in six different clades, closely related to Bartonella genotypes previously detected in D. rotundus and associated ectoparasites sampled in Latin America. On the other hand, the Bartonella rpoB genotypes clustered together with the ruminant species, B. schoenbuchensis and B. chomelii. The present study describes for the first time the molecular detection of Bartonella spp. in D. ecaudata bats. It also indicates that Bartonella spp. of vampire bats are genetically diverse and geographically widespread in Brazil.     

Survey of ticks of domestic dogs and cattle in three Caribbean islands

Ticks and the pathogens they transmit can cause high morbidity and mortality in domestic animals. As part of a larger study to determine the tick‐borne pathogens infesting domestic animals and wildlife, the aim of this study was to survey the tick species infesting the canine and cattle populations in Trinidad, Tobago and St. Lucia. A total of 1,990 ticks were collected off 179 dogs in Trinidad (n = 163) and Tobago (n = 16) between June 2016 and 2018. Ticks were also collected from cattle throughout Trinidad (n = 1,098), Tobago (n = 306) and St. Lucia (n = 176). Collected ticks were morphologically identified using standard taxonomic keys. Tick‐infested dogs were characterized as pets (n = 161) or hunting dogs (n = 18). Only two tick species, Rhipicephalus sanguineus (1,926; 96.8%) and Amblyomma ovale (64; 3.2%), were found on the dogs. A total of 169 (94.4%) dogs and 10 (5.6%) dogs were infested with R. sanguineus and A. ovale, respectively. Three dogs (1.7%) were infested with both tick species. Hunting dogs or those closely associated with them were infested with A. ovale. Rhipicephalus sanguineus was widely distributed throughout both islands, whereas A. ovale was restricted to small foci in three rural settlements in both Trinidad (n = 2) and Tobago (n = 1). Rhipicephalus (Boophilus) microplus (n = 1,404) was the only tick species found in cattle from Trinidad (n = 62) and Tobago (n = 20), whilst R. B. microplus (n = 171) and Amblyomma variegatum (n = 5) were found infesting 14 and two heads of cattle, respectively, in St. Lucia. These preliminary findings will aid in determining whether there are links between ticks and tick‐borne pathogens associated with domestic, wildlife species and humans and give further insight into the potential movement of ticks and their pathogens between the human, animal and tropical forest interface.     

Epidemiological Study of Lumpy Skin Disease Outbreaks in North‐western Iran

Lumpy skin disease (LSD) is a highly contagious transboundary disease of cattle with major economic losses. This study was undertaken to address the emergence and epidemiological features of LSD in four north‐western provinces of Iran. These provinces have extensive borders with others country including Iraq, Turkey, Azerbaijan and Armenia. A population of 683 cattle from 91 farms were examined during LSD outbreak in Iran during 2014–2016. The information of the farms including the population size, gender, age, vaccination status, clinical signs and the number of death because of LSD were recorded in the designed questionnaires. A number of 234 blood samples were collected randomly from animals with and without clinical signs of LSD. DNA was extracted from blood samples, and they were used for amplifying a fragment of 434 bp in size coupled with restriction fragment length polymorphism (RFLP) for molecular detection of lumpy skin disease virus (LSDV). The estimated prevalence, cumulative mortality and case fatality were 17.9%, 3.5% and 19.7%, respectively. There was no significant difference in occurrence of the disease between male and female cattle. LSD occurrence in age groups above 5 years old and below 6 months old showed highest and lowest relative frequencies, respectively. Vaccination was significantly decreased the occurrence of clinical disease. The developed PCR–RFLP technique was able to differentiate between LSDV, sheep pox virus (ShPV) and goat pox virus (GPV). It was concluded that LSD was entered into Iran probably from Iraq via uncontrolled animal movements along common land borders between two countries. Developed PCR–RFLP could be used as a rapid and inexpensive method for differentiating Capripoxviruses (CaPVs).     

Bovine anaplasmosis and tick‐borne pathogens in cattle of the Galapagos Islands

A cross‐sectional study was conducted to determine the species of Anaplasma spp. and estimate its prevalence in cattle of the three main cattle‐producing Galapagos Islands (Santa Cruz, San Cristóbal and Isabela) using indirect PCR assays, genetic sequencing and ELISA . Ticks were also collected from cattle and scanned for 47 tick‐borne pathogens in a 48 × 48 real‐time PCR chip. A mixed effects logistic regression was performed to identify potential risk factors explaining Anaplasma infection in cattle. A. phagocytophilum was not detected in any of the tested animals. Genetic sequencing allowed detection of A. platys ‐like strains in 11 (36.7%) of the 30 Anaplasma spp.‐positive samples analysed. A. marginale was widespread in the three islands with a global between‐herd prevalence of 100% [89; 100]_95%CI and a median within‐herd prevalence of 93%. A significant association was found between A. marginale infection and age with higher odds of being positive for adults (OR = 3.3 [1.2; 9.9]_{95% Bootstrap CI} ). All collected ticks were identified as Rhipicephalus microplus. A. marginale , Babesia bigemina , Borrelia theileri and Francisella ‐like endosymbiont were detected in tick pools. These results show that the Galapagos Islands are endemic for A. marginale .