Phenol/Chloroform/Proteinase K Multiplex PCR Epidemiology a b s t r a c t This study deals with the development and validation of an original PCR protocol to assess the presence of Fasciola hepatica in Galba truncatula its main intermediate host in Western Europe. In the present study two DNA extraction techniques are compared and a new multiplex PCR is described. The Chelex ® DNA extraction technique showed to be more appropriate than the classical Phenol/Chloroform/Proteinase K based method because of the absence of toxic organic solvent, shorter duration and lower cost, and a higher reproducibility regarding DNA concentrations and wavelength ratios. The multiplex PCR was set up to amplify the lymnaeid internal transcribed spacer 2 sequence (500-600 bp) that act as an internal control and a 124 bp Fasciola sp. sequence that is repeated more than 300,000 times in fluke whole genome. Ninety six snails were pooled and 6 snails (6.25%) found positive for Fasciola sp. The limit of detection is lower than the minimal biological infestation unit (one miracidium). DNA extracts from Paramphistomum daubneyi, Dicrocoelium lanceolatum, and Fascioloides magna did not cross react.

A molecular survey was undertaken to determine the presence of protozoal and bacterial organisms in 120 ticks and 87 blood samples collected from mammals in northeastern Algeria. Eight tick species were morphologically identified including 70 Rhipicephalus (Boophilus) annulatus,23 Rhipicephalus bursa, five Rhipicephalus sanguineus sensu lato, 11 Hyalomma impeltatum, five Hyalomma scupense, two Hyalommma marginatum, one Hyalomma anatolicum and three Ixodes ricinus. Quantitative PCR screening of the ticks showed that Theileria annulata, "Candidatus Ehrlichia urmitei", Theileria buffeli and Anaplasma platys were detected in Rh. annulatus. Rickettsia massiliae and Anaplasma ovis were detected in Rh. sanguineus s.l. and Rh. bursa. Rickettsia aeschlimannii was detected in Hy. marginatum, Hy. scupense and Hy. impeltatum. Finally, "Candidatus Rickettsia barbariae" was detected in Rh. bursa. In the screening blood samples, Theileria equi, T.annulata, T. buffeli, Babesia bovis, Anaplasma marginale, A. ovis and Borrelia spp. were detected in cattle. Theileria ovis, T. annulata, and A. ovis were detected in sheep. In addition, A. ovis and T. equi were detected in goats and equidea respectively. In this study, T. equi and "Candidatus Rickettsia barbariae" were identified for the first time in Algeria as well as potential new species of Ehrlichia and Anaplasma. Although molecular detection does not indicate vector/reservoir competence when investigating ticks removed from animals, this study expands the knowledge of the microorganisms detected in ticks in northeast of Algeria.

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is now routinely used for the rapid identification of microorganisms isolated from clinical samples and has been recently successfully applied to the identification of arthropods. In the present study, this proteomics tool was used to identify lice collected from livestock and poultry in Algeria. The MALDI-TOF MS spectra of 408 adult specimens were measured for 14 species, including Bovicola bovis, B. ovis, B. caprae, Haematopinus eurysternus, Linognathus africanus, L. vituli, Solenopotes capillatus, Menacanthus stramineus, Menopon gallinae, Chelopistes meleagridis, Goniocotes gallinae, Goniodes gigas, Lipeurus caponis and laboratory reared Pediculus humanus corporis. Good quality spectra were obtained for 305 samples. Spectral analysis revealed intra-species reproducibility and inter-species specificity that were consistent with the morphological classification. A blind test of 248 specimens was performed against the in-lab database upgraded with new spectra and validated using molecular tools. With identification percentages ranging from 76% to 100% alongside high identification scores (mean = 2.115), this study proposes MALDI-TOF MS as an effective tool for discriminating lice species.

Background and Aim:Lice are permanent ectoparasites, extremely specific to their hosts. Their great importance in veterinary medicine remain significant, they can cause their direct pathogenic actions like irritability, dermatitis, anemia, decreased weight gain, and milk production. The purpose of this work was to made the first time an inventory of mammalian lice in North-eastern Algeria.Materials and Methods:Our survey of lice infestation was conducted on several animal species from five provinces of North-eastern Algeria. A total of 57 cattle, 83 sheep, 77 goats, 111 wild boars, and 63 farmyard chickens were examined. The collection of lice was carried out much more in mammals and chickens during the winter period. Lice were collected either manually or using brushing and kept in flasks containing 70% ethanol. The identification of lice was achieved in the laboratory using a binocular loupe.Results:Concerning cattle, 63% and 27% of those examined subjects from Souk-Ahras and Guelma study areas, respectively, were carriers of lice. Damalinia bovis was the louse most frequently found on cattle in these two regions. Three other species were identified in Souk-Ahras: Haematopinus eurysternus (25%), Linognathus vituli (10%), and Solenopotes capillatus (5%). Regarding sheep, 39% and 24% of examined animals in Souk-Ahras and Guelma, were carrying lice. Damalinia ovis was the most frequently encountered lice on sheep in both regions. Linognathus ovillus also was identified in Souk-Ahras, representing 0.3% of the collected lice. Concerning goats, 53% and 30% of examined animals in Souk-Ahras and Guelma, were parasitized of lice. Two species of lice were found: Damalinia caprae and Linognathus africanus. For farmyard chickens, 69% and 100% of the farmyard chicken in Souk-Ahras and Mila were parasitized by lice, respectively. Menopon gallinae was the most frequently encountered louse in farmyard chicken in both regions. Eight other species were identified in Mila and four other species only in Souk-Ahras. Finally, 25% and 28% of the wild boars in Annaba and El Tarf were parasitized by lice, respectively. Haematopinus suis was the only species found on wild boars in both regions.Conclusion:These results are to be taken into account for lice control schemes and louse-borne diseases.

Free-range backyard chicken breeding is gaining popularity in Algerian rural regions. Due to the natural rearing conditions, chickens are exposed a wide range of microbes and ectoparasites. Currently, little is known about the variety of ectoparasites infesting free-range chickens in Algeria. The present study was conducted over a two year period on 169 backyard chicken, with the aim of identifying and estimating the prevalence of ectoparasites on different body parts of the chickens, and their immediate environments in northeast Algeria. In total, 9943 ectoparasites were identified, and the seven most dominant species were: Menopon gallinae (82.84%), Goniodes dissimilis (15.97%), Menacanthus stramineus (13.60%), Goniocotes gallinae (6.50%), Lipeurus caponis (5.23%), Argas persicus (9.46%) and Dermanyssus gallinae (1.18%). Menopon gallinae was the most frequent isolated ectoparasite throughout the year. Soft ticks Argas persicus were isolated during spring and summer, mainly from crevices and cracks, while the chicken red mite Dermanyssus gallinae was detected in autumn with a low load on bird body parts and a high abundance in nests. These results list the most abundant ectoparasites in backyard chickens, which could facilitate the improvement of rearing management.