Molecular prevalence and phylogenetic analysis of Theileria annulata and Trypanosoma evansi in cattle in Northern Tunisia.

The present study aimed to estimate the molecular prevalence of Theileria annulata and Trypanosoma evansi infection in cattle in Northern Tunisia. A total number of 96 cattle from five farms were evaluated. T. annulata and T. evansi prevalences were 61% [56/66] and 10% [7/13], respectively, at a confidence interval (CI) of 95%, while co-infection was present in 6% [4/8] of the tested animals at a CI of 95%. There was a significant correlation between age and the prevalence of T. annulata infection, whereas, there was no significant association shown with the age of cattle and T. evansi infection. Sequence and phylogenetic analyses showed that the T. annulata Tams1 gene and T. evansi ITS1 rDNA gene were highly conserved with 97.1-100% and 98.3-100% sequence identity, respectively.

Molecular detection and phylogenetic analyses of Toxoplasma gondii from naturally infected sheep in Northern and Central Tunisia.

Toxoplasmosis is a worldwide zoonosis caused by the parasitic protozoan Toxoplasma gondii. It can infect all warm-blooded vertebrate species and causes abortions and birth defects in pregnant women and pregnant ewes. The objective of this study was to estimate the prevalence of infection with T. gondii in sheep meat in the region of Sidi Bouzid (central Tunisia) and Beja (northern Tunisia), the realization of a descriptive study of risk factors and the phylogenetic analyses of T. gondii. Neck muscle samples were obtained from 174 ewes and ewe lamb slaughtered in Sidi Bouzid and 150 lambs slaughtered in Beja. DNA was extracted from the samples using the Wizard® genomic DNA purification kit. A nested PCR using two pairs of primers (NN 1 and NN2, Tg-NP1 and Tg-NP2) were used to detect infection with T. gondii, which was then confirmed by sequencing. Eight T. gondii amplicons were sequenced (accession number KT896498) and deposited in GenBank. The T. gondii amplicons showed 97-100% identities with GenBank sequences. A phylogenetic tree was then constructed. The nested PCR detected T. gondii DNA in 31% of animals tested in Sidi Bouzid and 32% of lambs tested in Beja. No significant difference in the prevalence of T. gondii infection was established between the two tested regions. In both regions, no significant variation of the infection depending on age, breed and locality was found.

Effect of the infection with the nematode Haemonchus contortus (Strongylida: Trichostrongylidae) on the haematological, biochemical, clinical and reproductive traits in rams.

This study aimed to investigate the effect of Haemonchus contortus infection on rams' haematological, biochemical and clinical parameters and reproductive performances. A total number of 12 Barbarine rams (control and infected) were included in the experiment. The infected group received 30 000 H. contortus third-stage larvae orally. Each ram's ejaculate was immediately evaluated for volume, sperm cell concentration and mortality rate. At the end of the experiment (day 82 post-infection), which lasted 89 days, serial blood samples were collected in order to assess plasma testosterone and luteinising hormone (LH) concentrations. There was an effect of time, infection and their interaction on haematological parameters (p < 0.001). In infected rams, haematocrit, red blood cell count and haemoglobin started to decrease from 21 days post-infection. There was an effect of time and infection for albumin. For total protein, only infection had a statistically significant effect. For glucose, only time had a statistically significant effect. Concentrations were significantly lower in infected rams compared to control animals. A significant effect of infection and time on sperm concentrations and sperm mortality was observed. The effect of infection appears in time for sperm concentrations at days 69 and 76 post-infection. Sperm mortality rate was significantly higher in infected animals at day 46 post-infection when compared to control group (p < 0.05). Finally, plasma testosterone traits (average concentration, cumulated levels during the sampling period and pulse frequency) were depressed in infected rams when compared to control counterparts; none of these endocrine traits were affected for plasma LH.

Molecular prevalence and genetic characterization of piroplasms in dogs from Tunisia.

In this study, the prevalence of piroplasms in dogs was assessed using polymerase chain reaction (PCR) to identify Babesia and Theileria species in 200 dogs from Northern and Central Tunisia between spring and autumn 2014. The overall molecular prevalence for piroplasms was 14·5% ± 0·05 (29/200); PCR detected 2 species, namely Babesia vogeli and Theileria annulata with an overall prevalence of 12·5 ± 0·04 and 2% ± 0·02, respectively. No differences in the molecular prevalences of B. vogeli were revealed for age and sex (P > 0·05). The molecular prevalence of B. vogeli was significantly higher in central Tunisia (26·5% ± 0·01) compared with the North (9·6% ± 0·04) (P 0·05). Comparison of the partial sequences of 18S rRNA and Tams 1 genes confirmed the presence of 2 novel B. vogeli and T. annulata genotypes. This is the first molecular detection of T. annulata and genetic characterization of dogs' piroplasms in Tunisia. Further studies are needed to better assess the epidemiological feature of piroplasms infection in North Africa.

Prevalence of Theileria and Babesia species in Tunisian sheep.

In this study, the prevalence of Theileria and Babesia species in sheep was assessed with Giemsastained blood smear examination and polymerase chain reaction to identify the different piroplasms in 270 sheep from three Tunisian bioclimatic zones (north, centre, and south). The overall infection prevalence by Babesia spp. and Theileria spp. in Giemsa-stained blood smears was 2.9% (8/270) and 4.8% (13/270) respectively. The molecular results showed that sheep were more often infected by Theileria ovis than Babesia ovis with an overall prevalence of 16.3% (44/270) and 7.8% (21/270) respectively (p = 0.01). The molecular prevalence by Babesia ovis was significantly higher in females than in males (p < 0.05). According to localities B. ovis was found exclusively in sheep from the centre of Tunisia (Kairouan) whereas Theileria ovis was found in all regions. Infections with T. ovis and B. ovis were confirmed by sequencing. The sequence of T. ovis in this study (accession numbers KM924442) falls into the same clade as T. ovis deposited in GenBank. The T. ovis amplicons (KM924442) showed 99%-100% identities with GenBank sequences. Moreover, comparison of the partial sequences of 18S rRNA gene of B. ovis described in this study (KP670199) revealed 99.4% similarity with B. ovis recently reported in northern Tunisia from sheep and goats. Three nucleotides were different at positions 73 (A/T), 417 (A/T), and 420 (G/T). It also had 99% identity with B. ovis from Spain, Turkey and Iraq. The results suggest a high T. ovis prevalence in Tunisia with a decreasing north-south gradient. This could be correlated to the vector tick distribution.

Molecular prevalence of Toxoplasma gondii DNA in goats' milk and seroprevalence in Northwest Tunisia.

Toxoplasmosis is a worldwide zoonosis with high impact on human and animal health. Consumption of unpasteurized milk is a risk factor of human toxoplasmosis. The aim of this study was to estimate the seroprevalence and molecular prevalence of T. gondii in goats' milk in Northwest of Tunisia (Jendouba Governorate). A total number of 77 blood samples were collected from six herds were screened with a commercial ELISA kit for T. gondii antibodies. For the same goats' samples, a nested PCR was performed to detect T. gondii DNA in milk. The seroprevalence of T. gondii infection was 31.2% (±0.05) while the molecular prevalence of this parasite in milk was estimated to 7.8% (±0.03). A very low value of kappa showed that there is not agreement between seroprevalence and parasite prevalence in milk. These results suggest that the consumption of raw milk from naturally infected goats is a potential source of human infection. An extension programme should be implemented to decrease related to goats' raw milk consumption.

First molecular isolation of Mycoplasma ovis from small ruminants in North Africa.

Eperythrozoonosis is a small ruminant disease caused by the bacterium Mycoplasma ovis (formerly known as Eperythrozoon ovis). Whilst acute infection in sheep may result in an anaemia and ill thrift syndrome, most animals do not develop clinical signs. Molecular methods were used to compare and evaluate the prevalence of infection with M. ovis in sheep and goats in Tunisia. A total of 739 whole blood samples from 573 sheep and 166 goats were tested for the M. ovis 16S rRNA gene using PCR. The overall prevalence was 6.28% ± 0.019 (36/573). Only sheep were infected with M. ovis (p < 0.001), and the prevalence was significantly higher in central Tunisia (29.2%) compared with other regions (p < 0.05). The prevalence revealed significant differences according to breed and bioclimatic zones (p < 0.001). Furthermore, the prevalence in young sheep (35/330; 10.6%) was higher than in adults (1/243; 0.41%) (p < 0.001). Only sheep of the Barbarine breed were infected, with a prevalence of 11.8% (p < 0.001). This is the first molecular study and genetic characterisation of M. ovis in North African sheep breeds.