Molecular characterization and phylogenetic analysis of Fasciola hepatica from high- plateau and steppe areas in Algeria.

A previous study based on mitochondrial DNA markers reported the presence of Fasciola hepatica in Algeria. However, a precise species identification is still required. In this report, a total of 68 Fasciola isolates, collected from high-plateau (Bordj-Bou-Arreridj) and steppe (Djelfa) areas of Algeria, were identified at the species level by multiplex PCR and PCR-restriction fragment length polymorphism (RFLP) for nuclear phosphoenolpyruvate carboxykinase (pepck) and DNA polymerase delta (pold), respectively. The result of the multiplex PCR conflicted with that of the PCR-RFLP; however, subsequent nucleotide sequencing of pepck clearly showed that all isolates should be classified as F. hepatica. The two mitochondrial markers, NADH dehydrogenase subunit I (nad1) and cytochrome c oxidase subunit 1 (cox1), revealed a close relationship between the parasite populations from the plateau and those from the steppe. A dispersal direction from the high plateau to the steppe was indicated because the former population was more diversified than the latter. Moreover, these populations were more closely related to populations from Spain than those from Egypt or Afghanistan. Given the population characteristic of F. hepatica in Spain and the history of cattle trade, it seems likely that the parasite was introduced to Algeria from Europe through a route across the Mediterranean Sea.

Frequency and genetic diversity of Echinococcus granulosus sensu stricto in sheep and cattle from the steppe region of Djelfa, Algeria.

Cystic echinococcosis (CE) of humans and animals is caused by various species of Echinococcus granulosus sensu lato. Of these, E. granulosus sensu stricto has the widest geographical distribution and is the most important agent of human cystic echinococcosis. Previous molecular studies showed that E. granulosus s.s. isolates from the Middle East and western Asia exhibit higher intraspecific diversity than those from other parts of the world, which led to hypotheses on the origin of the species in that region. However, various high-endemicity regions have not been sufficiently covered by such studies, including northern Africa as a well-known focus of this parasite. Here, we report data on the mitochondrial cox1 gene (1609bp) sequence diversity of E. granulosus s.s. from Algerian livestock. An abattoir survey of 1278 animals from the Algerian steppe region (Djelfa) resulted in CE prevalence of 13.9% in cattle (n = 266), 5.7% in sheep (n = 975), and 0% in goats (n = 37). All of 125 molecularly examined cyst isolates belonged to E. granulosus s.s. In total, 73 haplotypes were found, only five of which have been previously reported (from the Middle East and Australia). One haplotype sequence (EgAlg01X) was found to contain an insertion of three bases at the end of the gene. To the best of our knowledge, this has not been reported before for Echinococcus spp. Diversity values of our panel of Algerian samples were in the range of those that have been previously reported from the Middle East and far higher than those from elsewhere. This, together with the low number of shared haplotypes, indicates a more complex biogeographical history of this parasite than hitherto assumed.

Microsporidia and Cryptosporidium in horses and donkeys in Algeria: detection of a novel Cryptosporidium hominis subtype family (Ik) in a horse.

A total of 219 and 124 individual fecal samples of horses and donkeys, respectively, were screened for the presence of Cryptosporidium spp., Encephalitozoon spp., and Enterocytozoon bieneusi DNA by genus-specific nested PCR. Isolates were genotyped by sequence analysis of SSU rRNA, GP60, TRAP-C1, COWP, and HSP70 loci in Cryptosporidium, and the ITS region in microsporidia. Cryptosporidium spp. was detected on 3/18 horse farms and 1/15 farms where donkeys were kept. Overall, five (2.3%) horse and two (1.6%) donkey specimens were PCR positive for Cryptosporidium. Genotyping at SSU and GP60 loci revealed that three isolates from horses and donkeys were C. parvum subtype family IIaA16G1R1, one isolate from a horse was, C. muris RN66, and one isolate from a donkey was C. muris TS03. An isolate from a horse shared 99.4% and 99.3% similarity with Cryptosporidium hominis and C. cuniculus, respectively, at the SSU locus. This isolate shared 100% identity with C. hominis at the TRAP-C1, COWP, and HSP70 loci, and it was from the novel gp60 subtype family IkA15G1. Microsporidia were found on 6/18 horse and 2/15 donkey farms. E. bieneusi was identified in 6.8% (15/219) and 1.6% (2/124), and Encephalitozoon cuniculi was identified in 1.8% (4/219) and 1.6% (2/124), of horses and donkeys, respectively. Three genotypes of E. cuniculi (I, II and III) were detected in horses, and E. cuniculi genotype II was detected in donkeys. Four genotypes of E. bieneusi (horse1, horse 2, CZ3, D) were described in horses. An additional five horses and two donkeys were positive for E. bieneusi, but the isolated were not genotyped. Neither Cryptosporidium nor microsporidia prevalence were affected by sex, age, type of breeding, or whether the host was a horse or a donkey.

Equine cryptosporidial infection associated with Cryptosporidium hedgehog genotype in Algeria.

Faecal samples from two horse farms in Algeria keeping Arabian, Thoroughbred, and Barb horses were examined for the presence of Cryptosporidium in 2010-2011. A total of 138 faecal samples (16 from a farm keeping 50 animals and 122 from a farm with 267 horses) were screened for Cryptosporidium spp. infection using molecular tools. DNA was extracted from all samples. Nested PCR was performed to amplify fragments of the SSU rDNA and gp60 genes to determine the presence of Cryptosporidium species and genotypes. Sequence analyses of SSU and gp60 genes revealed four animals positive for the presence of subtype XIIIa A22R9 of the Cryptosporidium hedgehog genotype. The infections were not associated with diarrhoea. This study reports, for the first time, the occurrence of Cryptosporidium in Algeria and the first occurrence of the hedgehog genotype in horses. These findings support the potential role of infected horses in sylvatic-domestic transmission of Cryptosporidium.