Repeated Cross-Sectional Survey of Ectoparasites in Sheep from Central Tunisia: Does Low Prevalence Indicate Good Hygiene or Resistance to Ectoparasites?

Sheep ectoparasites such as chewing lice, fleas and ticks are serious constraints to sheep productivity and are the cause of skin lesions in animals that decrease their market value. This study aims at investigating the ectoparasite fauna infesting small ruminants in the district of Sidi Bouzid (central Tunisia). A total of 1243 Barbarine and Queue Fine de l'Ouest (QFO) sheep were examined every two months for one year. Of the total animals examined, 74 were infested by at least 1 parasite group (5.95%). Three ectoparasite groups were identified as Psoroptes ovis (0.48%; 6/1243), ticks (5.3%; n = 66/1243) and one specimen of Ctenocephalides canis (0.08%; n = 1/1243). The most abundant tick among the 358 specimens was Rhipicephalus sanguineus sensu lato (n = 337; 94.1%), followed by Hyalomma impeltatum (n = 7/358; 1.9%), H. dromedarii (n = 7/358; 1.9%), H. excavatum (n = 5/358; 1.4%) and only two specimens of H. scupense (n = 2/358; 0.55%). The sheep herds showed low infestation prevalence by ectoparasite over the year, with a significant difference according to the seasons (p < 0.05). A higher infestation prevalence was recorded in March (14.36%). Barbarine sheep breed showed significantly higher infestation prevalence (16.8%) compared to QFO (0.8%) (p < 0.01). There were no differences in infestation prevalence according to sex of the animal or age groups. Knowledge of the ectoparasite population harboured by sheep, its activity dynamics and risk factors is required to develop effective ectoparasite control options. The low prevalence of ectoparasite infestation in sheep reported here may be due to possible genetic resistance or simply to successful hygiene measures implemented by farmers.

In vitro infection of bovine erythrocytes with Theileria annulata merozoites as a key step in completing the T. annulata life cycle in vitro.

Theileria annulata is a protozoan parasite with a complex life cycle involving a bovine host and a tick vector. It is transmitted by Hyalomma ticks and is the causative agent of tropical theileriosis, a debilitating and often fatal disease in southern Europe, northern Africa and large parts of Asia. Understanding the biology of different life cycle stages is critical for the control of tropical theileriosis and requires the use of experimental animals which poses an ethical concern. We present for the first time the in vitro infection of red blood cells (RBCs) with T. annulata differentiated schizonts. The Ankara cell line of T. annulata was cultured at 41 °C for nine days to induce merogony and subsequently incubated with purified RBCs for one to three days. Percentage of parasitized erythrocyte (PPE) over the short culture period was estimated by Giemsa staining (0.007-0.01%), Flow cytometry activated sorting (FACS) (0.02-1.1%) and observation of FACS sorted cells by confocal microscopy (0.05-0.4%). There was a significant difference in the PPE between FACS and the two other techniques (one-way ANOVA followed by Tukey test, P = 0.004) but no significant difference was observed between the confocal imaging and Giemsa staining methods (ANOVA one-way followed by Tukey test, P = 0.06). Importantly, all three complementary methods confirmed the invasion of RBCs by T. annulata merozoites in vitro. Although the experimental conditions will require further optimization to increase the PPE, the in vitro infection of RBCs by T. annulata merozoites is pivotal in paving the way for the eventual completion of the T. annulata life cycle in vitro when combined with artificial tick feeding.

In vitro feeding of all life stages of two-host Hyalomma excavatum and Hyalomma scupense and three-host Hyalomma dromedarii ticks.

Ticks are blood-sucking ectoparasites and can transmit various pathogens of medical and veterinary relevance. The life cycle of ticks can be completed under laboratory conditions on experimental animals, but the artificial feeding of ticks has attracted increased interest as an alternative method. This study represents the first report on the successful in vitro feeding of all life stages of two-host tick species, Hyalomma scupense and Hyalomma excavatum, and the three-host tick Hyalomma dromedarii. The attachment and engorgement rates of adults were 84% (21/25) and 76% (19/25) for H. scupense females. For adult H. excavatum and H. dromedarii, 70% (21/30) and 34.4% (11/32) of the females attached and all attached females successfully fed to repletion. The oviposition rates of the artificially fed females were 36.4%, 57.1% and 63.1% for H. dromedarii, H. excavatum and H. scupense, respectively, with a reproductive efficiency index varying between 44.3 and 60.7%. For the larvae, the attachment and engorgement rates were 44.2% (313/708) and 42.8% (303/708) for H. dromedarii, 70.5% (129/183) and 56.8% (104/183) for H. excavatum and 92.6% (113/122) and 55.7% (68/122) for H. scupense. The attachment and engorgement rates for the nymphs were 90.2% (129/143) and 47.6% (68/143) for H. dromedarii, 66.7% (34/51) and 41.2% (21/51) for H. excavatum, and 44.1% (30/68) and 36.8% (25/68) for H. scupense. Molting rates of the immature stages varied between 71.3% (216/303) and 100% (68/68) for the larvae and between 61.9% (13/21) and 96% (24/25) for the nymphs. The successful in vitro feeding of all stages of the three Hyalomma species makes this method a valuable tool for tick research, with potential applications in studies on the pathogens transmitted by these tick species such as Theileria annulata.

Dual RNA-seq to catalogue host and parasite gene expression changes associated with virulence of T. annulata-transformed bovine leukocytes: towards identification of attenuation biomarkers.

The apicomplexan parasite Theileria annulata is transmitted by Hyalomma ticks and causes an acute lymphoproliferative disease that is invariably lethal in exotic cattle breeds. The unique ability of the schizont stage of T. annulata to transform infected leukocytes to a cancer-like phenotype and the simplicity of culturing and passaging T. annulata-transformed cells in vitro have been explored for live vaccine development by attenuating the transformed cells using lengthy serial propagation in vitro. The empirical in vivo evaluation of attenuation required for each batch of long-term cultured cells is a major constraint since it is resource intensive and raises ethical issues regarding animal welfare. As yet, the molecular mechanisms underlying attenuation are not well understood. Characteristic changes in gene expression brought about by attenuation are likely to aid in the identification of novel biomarkers for attenuation. We set out to undertake a comparative transcriptome analysis of attenuated (passage 296) and virulent (passage 26) bovine leukocytes infected with a Tunisian strain of T. annulata termed Beja. RNA-seq was used to analyse gene expression profiles and the relative expression levels of selected genes were verified by real-time quantitative PCR (RT-qPCR) analysis. Among the 3538 T. annulata genes analysed, 214 were significantly differentially expressed, of which 149 genes were up-regulated and 65 down-regulated. Functional annotation of differentially expressed T. annulata genes revealed four broad categories of metabolic pathways: carbon metabolism, oxidative phosphorylation, protein processing in the endoplasmic reticulum and biosynthesis of secondary metabolites. It is interesting to note that of the top 40 genes that showed altered expression, 13 were predicted to contain a signal peptide and/or at least one transmembrane domain, suggesting possible involvement in host-parasite interaction. Of the 16,514 bovine transcripts, 284 and 277 showed up-regulated and down-regulated expression, respectively. These were assigned to functional categories relevant to cell surface, tissue morphogenesis and regulation of cell adhesion, regulation of leucocyte, lymphocyte and cell activation. The genetic alterations acquired during attenuation that we have catalogued herein, as well as the accompanying in silico functional characterization, do not only improve understanding of the attenuation process, but can also be exploited by studies aimed at identifying attenuation biomarkers across different cell lines focusing on some host and parasite genes that have been highlighted in this study, such as bovine genes (CD69, ZNF618, LPAR3, and APOL3) and parasite genes such as TA03875.

Genetic diversity of Hyalomma marginatum in Tunisia is not influenced by the bio-climate.

Ticks are important ectoparasites responsible for the transmission of several pathogens with significant medical, veterinary, and economic impacts. Climate and social changes have generated substantial changes in ticks' distribution, abundance, and activity patterns, including ticks belonging to the Hyalomma marginatum species. Knowledge on the genetic structure and dynamics of H. marginatum populations might contribute to a better understanding of their current and future evolution under the effects of anthropogenic factors and eco-climatic changes. In the present study, we investigated the genetic structure and phylogenetic distribution of H. marginatum across three bioclimatic regions in Tunisia using two mitochondrial markers (16S and 12S rRNA). The molecular investigations were based on 47 adult H. marginatum ticks collected from humid, upper semi-arid, and sub-humid regions of Tunisia. Our results revealed a genetic diversity of 0.278% and 0.809% using the 16S and 12S markers, respectively. The low genetic diversity that we observed raises the hypothesis of a bottleneck event occasioned by a reduction in the size of the tick population under the effects of environmental factors and/or human activities. This hypothesis is supported by the population's demographic history analysis, which revealed a clear deviation from neutrality and supports the occurrence of a bottleneck event followed by a demographic expansion. The fact that most 16S and 12S variability was present in the ticks from the humid bioclimatic zone may suggest that those ticks represent the ancestral population. Overall, the analysis has shown that the phylogenetic clusters do not correspond to the bioclimatic zones.

Epidemiological situation of bovine tropical theileriosis in an arid region in central Tunisia with a phylogenetic analysis of Theileria annulata.

BACKGROUND: Tropical theileriosis, Theileria annulata infection, is the most prevalent summer disease in Tunisia. It is transmitted by Hyalomma scupense, a two-host tick known to be endophilic. OBJECTIVES: The present study aimed to estimate the infection prevalence of cattle by T. annulata in two districts from central Tunisia. METHODS: Blood samples collected from 270 Holstein cattle from the Sidi Bouzid (140 samples) and Kasserine districts (130 samples) were analysed by Giemsa staining and T. annulata-specific PCR. RESULTS: In both regions, PCR revealed a prevalence of 32.6%. This was significantly higher than the 6.3% prevalence obtained by Giemsa staining blood smears (p < 0.001). Giemsa staining also revealed a low parasitaemia of 0.05%. The PCR-based prevalence was not statistically different between the two districts (31.4 ± 0.04 and 33.8 ± 0.04% in Sidi Bouzid and Kasserine districts, respectively, p = 0.6). On the contrary, the results of blood smear examination (2.85 and 10% in Sidi Bouzid and Kasserine, respectively) differed significantly between the two sampling sites (p = 0.01). There was no evidence of a statistically significant difference between the overall molecular infection prevalence when the samples were segregated based on animals' age or gender (p = 0.1 and 0.2, respectively) and a similar trend was observed for Giemsa staining. Ten PCR amplicons of the Tams1 gene (721 bp) were subsequently sequenced from the two regions. The phylogenetic analyses showed 100% similarity between all sequences. The unique conserved Tams1 sequence was deposited in GenBank under the accession number OP428816 and used to infer its phylogenetic relationships with those available in the GenBank repository. CONCLUSIONS: This is the first report of the presence of T. annulata in this region of central Tunisia which has no history of tropical theileriosis. Priority areas for future studies include understanding the origin of these T. annulata-positive animals in a region where the presence of a known natural vector tick, H. scupense, has not been reported. Given that the disease severely constrains cattle productivity, it would also be worthwhile to investigate if other potential vectors for T. annulata, such as Hyalomma dromedarii, are present in the arid regions.

The antigen recognition portion of African buffalo class I MHC is highly polymorphic, consistent with a complex pathogen challenge environment, and the 3' region suggests distinct haplotype configurations.

African buffalo (Syncerus caffer) have been distinct from the Auroch lineage leading to domestic cattle for 5 million years, and are reservoirs of multiple pathogens, that affect introduced domestic cattle. To date, there has been no analysis of the class I MHC locus in African buffalo. We present the first data on African buffalo class I MHC, which demonstrates that gene and predicted protein coding sequences are approximately 86-87% similar to that of African domestic cattle in the peptide binding region. The study also shows concordance in the distribution of codons with elevated posterior probabilities of positive selection in the buffalo class I MHC and known antigen binding sites in cattle. Overall, the diversity in buffalo class I sequences appears greater than that in cattle, perhaps related to a more complex pathogen challenge environment in Africa. However, application of NetMHCpan suggested broad clustering of peptide binding specificities between buffalo and cattle. Furthermore, in the case of at least 20 alleles, critical peptide-binding residues appear to be conserved with those of cattle, including at secondary anchor residues. Alleles with six different length transmembrane regions were detected. This preliminary analysis suggests that like cattle, but unlike most other mammals, African buffalo appears to exhibit configuration (haplotype) variation in which the loci are expressed in distinct combinations.

Preliminary study on the seasonal questing of Ixodes ricinus group ticks in Ain Draham forest (north-western Tunisia) with analyses of their phylogenetic diversity.

The present study aimed to investigate the activity dynamics of Ixodes ricinus group ticks in a forest located in north-western Tunisia (Aïn Draham, Jendouba District) and assess the variation of abiotic factors (temperature, Normalized Difference Vegetation Index and relative humidity) during one year survey from September 2016 to August 2017 using the dragging sampling method. A total of 116 questing ticks was collected from the vegetation consisting of 47 adults (19 females and 28 males, sex ratio M:F = 1.47), 45 nymphs and 24 larvae representing 40.5, 38.8 and 20.7% of the total collected specimens, respectively. Adult I. ricinus were collected during October-May, nymphs during May-August and larvae during July-September. There were statistically significant correlations between adult tick numbers and mean daily relative humidity (Pearson r = 0.77; p = 0.003) and mean daily temperature (r = -0.74; p = 0.006). The comparison of 16S rDNA sequences from 20 adult ticks of approximately 444 bp length showed variability among 11 sequences. There was a low genetic variability (<1%) among the I. ricinus isolates collected from the forest. The amplicons showed >99% identity with I. ricinus and Ixodes inopinatus sequences from different countries and published in GenBank. These results should be complemented by further surveys in other Tunisian regions to better understand the influence of environmental factors on the biology of I. ricinus and the occurrence of sympatric I. inopinatus ticks. Different molecular markers should be used for better understanding of their taxonomic status.

Sequence polymorphisms in a Theileria annulata surface protein (TaSP) known to augment the immunity induced by live attenuated cell line vaccine.

Theileria annulata is a tick-borne protozoan causing tropical theileriosis in cattle. The use of attenuated cell line vaccines in combination with subunit vaccines has been relatively successful as a control method, as exemplified by a recent study in which immunization with a local cell line followed by booster vaccinations with recombinant T. annulata surface protein (TaSP) resulted in 100% protection upon field challenge in Sudan. However, these findings cannot be directly extrapolated to other countries as culture-attenuated live vaccines are generated using local strains and no systematic evaluation of genotype differences between countries has been undertaken. In this study, we sequenced the TaSP gene from T. annulata cell lines and field isolates from Tunisia (n = 28) and compared them to genotypes from Sudan (n = 25) and Morocco (n = 1; AJ316259.1). Our analyses revealed 20 unique TaSP genotypes in the Tunisian samples, which were all novel but similar to genotypes found in Asia. The impact of these polymorphisms on the ability of the TaSP antigen to boost the immunity engendered by live cell line vaccines, especially in Tunisia where studies with TaSP have not been conducted, remains to be examined. Interestingly, phylogenetic analyses of publicly available TaSP sequences resolved the sequences into two clusters with no correlation to the geographical origin of the isolates. The availability of candidate vaccines that were recently attenuated using local strains from Sudan, Tunisia, Egypt and Morocco should be exploited to generate a comprehensive catalogue of genetic variation across this regional collection of attenuated live vaccines.

Cultivation, cryopreservation and resuscitation of Theileria annulata transformed cells in serum-free media.

Introduction: Tropical theileriosis is a protozoan disease caused by Theileria annulata that affects cattle in Northern Africa, the Middle East and Asia where vector ticks of the genus Hyalomma occur. Various measures are applied to control the disease, including vaccination with attenuated T. annulata schizonts. Cultivation of T. annulata schizonts is mainly conducted in media containing Fetal Bovine Serum (FBS), which has some disadvantages such as costs, batch- to-batch variation and ethical concerns. Methods: In this study, we conducted three experiments to evaluate the ability of (1) T. annulata strains grown in RPMI with 10% FBS (RPMI-FBS) to adapt and grow in serum-free media (i.e., HL-1, RPMI without FBS supplementation, ISF-1, and M199), (2) a T. annulata strain grown in ISF-1 and subsequently frozen in this medium to grow in ISF-1 again after long-term storage in liquid nitrogen, and (3) a T. annulata strain freshly isolated from infected bovine lymphocytes to growin ISF-1, also after cryopreservation. Cell numbers, schizont index, the viability and generation doubling time were calculated in all experiments. Results and discussion: In the first experiment, the Hessiene and Beja cell lines from Tunisia previously cultivated in RPMI-FBS and adapted to serum-free media continued to grow significantly better in RPMI-FBS compared to the serum-freemedia. In the second experiment, a Tunisian cell line (Hessiene) cryopreserved in ISF-1 with 5%[v/v] dimethylsulfoxide (DMSO) grewbetter after thawing in RPMI-FBS compared to ISF-1 with a highly significant difference in cell growth (p < 0.001), whereas the third experiment showed that the Ankara cell line had similar growth characteristics in both RPMI-FBS and ISF-1 before and after thawing, with a shorter generation doubling time in ISF-1 than in RPMI-FBS (p = 0.23). Our findings suggest that freshly isolated cells can be propagated, frozen and thawed in serum-free media such as ISF-1, but once cells are adapted to cultivation in the presence of FBS or resuscitated from frozen storage, propagation in serum-free media may not perform as well as cultivation in RPMI-FBS.

Phenology and phylogeny of Hyalomma spp. ticks infesting one-humped camels (Camelus dromedarius) in the Tunisian Saharan bioclimatic zone.

In this study, we report the results of a survey of Hyalomma ticks infesting one-humped camels in southern Tunisia. Examinations were conducted every second or third month on 406 camels in Tataouine district from April 2018 to October 2019. A total of 1902 ticks belonging to the genus Hyalomma were collected. The ticks were identified as adult H. impeltatum (41.1%; n = 782), H. dromedarii (32.9%; n = 626), H. excavatum (25.9%; n = 493), and H. marginatum for a single specimen. Although the camels were infested by ticks throughout the year, the highest overall infestation prevalence was observed in April 2018 (p < 0.01). The overall infestation intensity varied between 2.7 and 7.4 ticks/animal. There were no statistically significant differences in tick infestation prevalence based on age categories of the camels, and the overall infestation prevalence was between 82.7% and 97.4%. Female camels were significantly more infested with ticks (88.3%) than males (65.5%) (p < 0.01). The infestation prevalence of camels varied significantly according to the region where sampling took place (p < 0.01), but no correlations were found with abiotic factors. The preferred attachment sites for adult Hyalomma ticks were the sternum (38.3%; n = 729/1902), around the anus (36.2%; n = 689/1902), udder (18.4%; n = 350/1902), and inner thigh (6.9%; n = 132/1902). Morphological classification of ticks was corroborated by sequencing the cytochrome c oxidase I (Cox1) and 16S rDNA genes, and these sequences were also used to infer phylogenetic relationships. A single H. dromedarii seemed to be a natural hybrid with H. rufipes. More attention should be devoted by the veterinary services to the infestation of camels by ticks.

TITLE: Phénologie et phylogénie des tiques Hyalomma spp. infestant les dromadaires (Camelus dromedarius) dans la zone bioclimatique saharienne tunisienne. ABSTRACT: Dans cette étude, les résultats d'une enquête concernant les tiques Hyalomma infestant les dromadaires dans le sud de la Tunisie sont présentés. Des examens ont été menés tous les deux ou trois mois sur 406 dromadaires dans le district de Tataouine entre avril 2018 et octobre 2019. Au total, 1902 tiques appartenant au genre Hyalomma ont été collectées. Les tiques adultes ont été identifiées comme H. impeltatum (41,1 % ; n = 782), H. dromedarii (32,9 % ; n = 626), H. excavatum (25,9 % ; n = 493) et un seul spécimen de H. marginatum. Bien que les dromadaires aient été infestés par les tiques tout au long de l'année, la prévalence globale d'infestation la plus élevée a été observée en avril 2018 (p < 0,01). L'intensité globale d'infestation variait entre 2,7 et 7,4 tiques/animal. Il n'y avait pas de différence statistiquement significative de l'infestation par les tiques en fonction des catégories d'âge des dromadaires, et la prévalence globale d'infestation se situait entre 82,7 et 97,4 %. Les dromadaires femelles étaient significativement plus infestés par les tiques (88,3 %) que les mâles (65,5 %) (p < 0,01). La prévalence d'infestation variait significativement en fonction des régions (p < 0.01) mais il n'y avait pas de corrélation avec les facteurs abiotiques. Les sites de fixation préférés des tiques Hyalomma adultes étaient le sternum (38,3 % ; n = 729/1902), autour de l'anus (36,2 % ; n = 689/1902), la mamelle (18,4 % ; n = 350/1902) et la face interne de la cuisse (6,9 % ; n = 132/1902). La classification morphologique a été corroborée par le séquençage des gènes de l'ADNr du cytochrome c oxydase I (Cox1) et du 16S, et ces séquences ont également été utilisées pour déduire les relations phylogénétiques. Un seul spécimen de H. dromedarii semblait être un hybride naturel avec H. rufipes. Une plus grande attention doit être accordée par les services vétérinaires quant à l'infestation des dromadaires par les tiques.

Epidemiological investigation of Crimean-Congo haemorrhagic fever virus infection among the one-humped camels (Camelus dromedarius) in southern Tunisia.

Crimean-Congo haemorrhagic fever is a viral tick-borne zoonotic disease caused by a Nairovirus, Crimean-Congo haemorrhagic fever virus (CCHFV). The present survey aimed to determine the exposure of one-humped camels (Camelus dromedarius) from southern Tunisia to CCHFV. A total of 273 sera from extensively reared camels were collected from Tataouine district, Tunisia, and tested by CCHFV-specific enzyme linked immunosorbent assays. By combining the results of three serological tests, the overall seroprevalence of CCHFV was estimated as 89.7% (245/273). No viral RNA was detected from camel sera using quantitative real-time PCR (RT-qPCR). A total of 165 ticks were collected from camels and tested with RT-qPCR, and only one Hyalomma impeltatum tick was positive for virus RNA.

Unique Mitochondrial Single Nucleotide Polymorphisms Demonstrate Resolution Potential to Discriminate Theileria parva Vaccine and Buffalo-Derived Strains.

Distinct pathogenic and epidemiological features underlie different Theileria parva strains resulting in different clinical manifestations of East Coast Fever and Corridor Disease in susceptible cattle. Unclear delineation of these strains limits the control of these diseases in endemic areas. Hence, an accurate characterization of strains can improve the treatment and prevention approaches as well as investigate their origin. Here, we describe a set of single nucleotide polymorphisms (SNPs) based on 13 near-complete mitogenomes of T. parva strains originating from East and Southern Africa, including the live vaccine stock strains. We identified 11 SNPs that are non-preferentially distributed within the coding and non-coding regions, all of which are synonymous except for two within the cytochrome b gene of buffalo-derived strains. Our analysis ascertains haplotype-specific mutations that segregate the different vaccine and the buffalo-derived strains except T. parva-Muguga and Serengeti-transformed strains suggesting a shared lineage between the latter two vaccine strains. Phylogenetic analyses including the mitogenomes of other Theileria species: T. annulata, T. taurotragi, and T. lestoquardi, with the latter two sequenced in this study for the first time, were congruent with nuclear-encoded genes. Importantly, we describe seven T. parva haplotypes characterized by synonymous SNPs and parsimony-informative characters with the other three transforming species mitogenomes. We anticipate that tracking T. parva mitochondrial haplotypes from this study will provide insight into the parasite's epidemiological dynamics and underpin current control efforts.

Current status of tropical theileriosis in Northern Africa: A review of recent epidemiological investigations and implications for control.

Tropical theileriosis caused by the apicomplexan hemoparasite Theileria annulata is a tick-borne disease that constraints livestock production in parts of Europe, Asia and Africa. Four Hyalomma tick species transmit T. annulata in at least eight Africa countries (Mauritania, Morocco, Algeria, Tunisia, Egypt, Sudan, South Sudan and Ethiopia). The two dominant T. annulata vector ticks present in Africa, H. scupense and H. anatolicum, underlie two different patterns of transmission, which in turn greatly influence the epidemiology of tropical theileriosis. H. dromedarii and H. lusitanicum are also capable of transmitting T. annulata in North Africa, but their roles are associated with specific production systems and agro-ecological contexts. The emergence of resistance to the most widely used theilericidal compound, buparvaquone, continues to limit the effectiveness of chemotherapy. In addition, acaricide use is increasingly becoming unsustainable. Deployable T. annulata attenuated live vaccines established from local strains in Tunisia, Sudan and Egypt are available, and recent work has indicated that these vaccines can be protective under conditions of natural transmission. However, vaccination programmes may vary over space and time due to differences in the prevalence of disease amongst cattle populations, as well seasonal variation in vector activity. We review recent descriptive and analytical surveys on the epidemiology of T. annulata infection with reference to (a) demographic aspects such as breeds and ages of cattle herds previously exposed to distinct T. annulata infection pressures and (b) seasonal dynamics of tick activity and disease transmission. We then discuss how the wider endemic patterns that we delineate can underpin the development and execution of future vaccination programmes. We also outline options for integrated control measures targeting tick vectors and husbandry practices.

Differences in tick infestation of Tunisian sheep breeds.

Different infestation patterns by ixodid ticks were studied in three sheep breeds in Tunisia: Barbarine, Queue Fine de l'Ouest and their cross-bred animals. During one year, 700 sheep were monitored and examined for tick infestation. A total of 722 ticks were collected from sheep ears. The most frequent tick species was by far Rhipicephalus sanguineus sensu lato (99%) and there were few specimens of Rhipicephalus bursa (1%) (p < 0.001). Overall infestation prevalence was estimated at 10.4%. The lowest infestation prevalence was in Barbarine sheep (7.3%), followed by Queue Fine de l'Ouest (16.7%) and the highest prevalence was in cross-bred sheep (19.1%) (p < 0.001). Mean overall infestation intensity was 1.6 ticks/sheep: lowest in Barbarine (1.4), followed by Queue Fine de l'Ouest (1.7) and cross-bred sheep (1.8). Similarly, abundance was lowest in Barbarine sheep (0.1), and was 0.3 in Queue Fine de l'Ouest and cross-bred animals. The results demonstrated a reduced infestation, possibly due to reduced attractiveness and/or increased resistance to tick infestation, of the Barbarine breed compared with the other two breeds. Further behavioural, genetic and molecular studies are needed to explain the mechanisms for the lower infestation indicators.