First molecular characterization of Blastocystis subtypes from animals and animal-keepers stool in Algeria.

Blastocystis sp. is one of the most common enteric parasites found in humans and many non-human hosts. It is an anaerobic protozoan that belongs to the group of Stramenopiles. Based on phylogenetic analysis of ribosomal DNA genes, at least 17 subtypes (ST1-ST17) are described. The aim of this study was to identify and characterize Blastocystis sp. in stool samples from various animal groups and animal-keepers. Overall, 29/70 (41.43%) animals and 7/60 (11.66%) humans sampled were positive for Blastocystis sp. using microscopy. The sequencing of the partial 18S small subunit ribosomal DNA gene (SSU rDNA) revealed the presence of five haplotypes corresponding to ST2 and ST3 in humans, and ST2, ST3, ST7, and ST10 in animals. This is the first report of Blastocystis subtypes in animals in Algeria.

Genetically 'pure' Fasciola gigantica discovered in Algeria: DNA multimarker characterization, trans-Saharan introduction from a Sahel origin and spreading risk into north-western Maghreb countries.

Fascioliasis is a freshwater snail-borne zoonotic helminth disease caused by two species of trematodes: Fasciola hepatica of almost worldwide distribution and the more pathogenic F. gigantica restricted to parts of Asia and most of Africa. Of high pathological impact in ruminants, it underlies large livestock husbandry losses. Fascioliasis is moreover of high public health importance and accordingly included within the main neglected tropical diseases by WHO. Additionally, this is an emerging disease due to influences of climate and global changes. In Africa, F. gigantica is distributed throughout almost the whole continent except in the north-western Maghreb countries of Morocco, Algeria and Tunisia where only F. hepatica is present. The present study concerns the DNA multimarker characterization of the first finding of F. gigantica in sheep in Algeria by the complete sequences of rDNA ITS-1 and ITS-2 and mtDNA cox1 and nad1 genes. Sequence comparisons and network analyses show sequence identities and similarities suggesting a south-north trans-Saharan geographical origin, with introduction from Ghana, through the Sahel countries of Burkina Faso and Mali into Algeria. This way perfectly fits with nomadic pastoralism according to interconnecting intranational and transborder herd transhumance routes traditionally followed in this western part of Africa from very long ago. The risk for further spread throughout the three north-western Maghreb countries is multidisciplinarily analysed, mainly considering the present extensive motorization of the intranational transhumance system in Algeria, the lymnaeid snail vector species present throughout the north-western Maghreb, the increasing demand for animal products in the growing cities of northern Algeria and the continued human infection reports. Control measures should assure making anti-fasciolid drugs available and affordable for herders from the beginning and along their transhumant routes and include diffusion and rules within the regional regulatory framework about the need for herd treatments.

Development of analytical methods GC-MS vs LC-UV for the serum monitoring of an inflammatory glycotoxin (methylglyoxal): A new biomarker of bovine hepatobiliary distomatosis.

Two analytical methods; high performance liquid chromatography and gas chromatography were used to determine the content of 2-methylquinoxaline, a methylglyoxal-derived agent in sera from cattle with fascioliasis. Methylglyoxal is a highly mutagenic and cytotoxic reactive dicarbonyl compound formed by non-enzymatic fragmentation of triose phosphate GAP and DHAP during glycolysis which regularly contributes to repositioning the energetic balance between physiological and pathological situations. The aim of this study was to propose the MGO as a new biomarker in the bovine fasciolosis. Strongly infected animals showed a correlation between the relatively high levels of Fasciola hepatica anti-f2 antibody and methylglyoxal compared to unharmed animals. Also, an acute hyperglycemia was recorded and closely related to hepatic parenchyma hyperplasia, inflammation, bile ducts obstruction and scléro-fibrous foci formation.Unlike HPLC, which has shown analytical flaws and irregularities, GC-MS remains an excellent diagnostic tool for detecting and quantifying methylglyoxal in biological fluids. The developed method has been validated under FDA guidelines. A full scan-range was set from m/z 39 to 144/999 and the molecular weight of the 2-methylquinoxaline was identified according to NIST Database and ES. Methylglyoxal was the only analyte successfully quantified in a relatively short run time. It was linear over a concentration range of 0.057-5.7  µg.ml-1with mean recoveries and RSD of 118% and 3.63% respectively. The intra and inter-day assays were satisfying and not exceed 3.00%. Results reflect the degree of precision of our method and indicate that MGO was an important contributor to understand the hepatic failure independently of other serum markers.

Seroprevalence and risk factors of Toxoplasma gondii infection in rabbit of local Algerian population.

AIM: The objective of this investigation was to determine the seroprevalence and identify the risk factors of Toxoplasma gondii infection in the rabbit of the local Algerian population from five districts of Northern Algeria. MATERIALS AND METHODS: Blood samples of 350 rabbits were collected and analyzed for the presence of anti-T. gondii immunoglobulin G antibodies using the indirect enzyme-linked immunosorbent assay. Additional data concerning the farms and management practices were obtained through a questionnaire used in surveys and interviews. RESULTS: The overall seroprevalence was 14.6% (51/350). The seroprevalence was significantly higher in outdoor rearing farms and was linked to the presence of animals from other species on the farm. A higher seroprevalence was found in older animals compared to younger ones. No difference in seroprevalence was noted with respect to the origin or gender of animals, type of cage, feed and water sources, presence of cats in the vicinity, hygiene status, or season. CONCLUSION: This study has shown that T. gondii prevalence in rabbits of the local population is relevant and may have important implications for public health in rural areas.

A post-mortem study of bovine fasciolosis in the Mitidja (north center of Algeria): prevalence, risk factors, and comparison of diagnostic methods.

The present study was conducted to determine the prevalence of fasciolosis and investigate the associated risk factors (sex, age, and season) in slaughtered cattle of the Mitidja area. This survey also aimed to compare three diagnostic methods: liver inspection, microscopic bile examination, and ELISA test. Liver and bile of 1400 cattle were examined for the presence of adult and eggs of Fasciola hepatica, respectively. Based on the results obtained by liver and bile examination, a subset of 206 cattle from the 1400 was selected for antibodies detection using the ELISA test. Liver inspection and microscopic bile examination showed prevalences of 2.86% and 5.50%, respectively. The difference between the two methods was highly significant (p < 0.001). The overall prevalence of bovine fasciolosis found by both methods was 6.07%. The disease was more prevalent in females (11.96%) than in males (2.43%), and in older cattle (12.30%) than in younger ones (2.36%). The ELISA test showed some conflicting results: 23.61% of cattle previously diagnosed positive with liver inspection and bile examination were found seronegative, and 3.73% of cattle previously diagnosed negative were found to be seropositive. The sensitivity and specificity of microscopic bile examination were respectively 80% (CI 65.2-89.5%) and 96.7% (CI 95.6-97.5%) as compared with the liver inspection. Likewise, this same method has a sensitivity of 81.36% (CI 69.6-89.3%) and a specificity of 88.97% (CI 82.8-93.1%) as compared with the ELISA test. Our results show that microscopic bile examination is able to detect more infected cattle than liver inspection. In addition to being simple, fast, inexpensive, sensitive, and specific, it is an effective method for the diagnosis of fasciolosis.

New insight into genetic variation and haplotype diversity of Fasciola hepatica from Algeria.

The liver fluke Fasciola hepatica is the main cause of fasciolosis in North Africa leading to significant economic losses and public health problems. In this study, the ribosomal internal transcribed spacer (ITS), cytochrome c oxidase I (COI), the mitochondrial region spanning the COI-trnT-rrnL, and the NADH dehydrogenase subunit I (NADI) markers were used to characterize Fasciola flukes from Algeria. Fasciola appeared widespread from the east to the west of Algeria. Among 1701 sampled cattle from 8 Algerian provinces, 5% were infected. Using morphological and morphometric analysis, one morphotype of Fasciola was observed. Nuclear ITS marker indicated that all collected flukes belong to F. hepatica. Multiple alignments of ITS dataset revealed two haplotypes, one described here for the first time. Analysis of molecular variance (AMOVA) of mitochondrial markers revealed weak population structure in Algeria. Mismatch distributions, neutrality tests, and median-joining network analysis all were compatible with a recent expansion of Algerian F. hepatica population. Fasciolosis appeared common in Algerian cattle, it seems that the absence of control strategy coupled to the favorable Mediterranean climate may lead to a reconstruction and dispersion of its populations. This study provides important results concerning the genetic characterization and variability of F. hepatica in Algeria as well as the significant role of cattle importation in shaping its dispersal route worldwide.

Evaluation of Fasciola hepatica Infections in Cattle in Northeastern Algeria and the Effects on Both Enzyme and Hepatic Damage, Confirmed by Scanning Electron Microscopy.

PURPOSE: The aim of our study is to establish the presence of Fasciola hepatica on farms in northeastern Algeria. METHODS: 143 blood and coprological samples of 15 males and 128 females of different breeds and ages were analysed. RESULTS: Our study indicates a heterogeneous level of the anti-f2 antibodies to Fasciola hepatica in response to the infection. The overall seroprevalence was about 22.37%, and 9 out of 13 investigated farms were infected, with rates varying from 5.88% to 70%. To explain the intrinsic variability of the infection, we identified age, sex and breed as potential risk factors. Based on this, we collected information about their relevance. There was a significant difference for age (p = 0.018) and coprology (p < 0.0001). Independently, sex and breed had no impact on the infection, although males were more infected (27%) than females (22%). Of the five investigated breeds, Holstein cattle (31.11%) were most affected, followed by Montbeliard (20.27%) and crossed breed (13.64%). Multivariate comparisons showed that the presence of faecal eggs reflects the active infectious status of cattle (p < 0.0001), while age (p = 0.011) and sex (p = 0.040) significantly impact the chance of acquiring the infection. To evaluate liver parenchyma integrity and its functionality, hepatic enzymes were examined and showed relatively low levels of aminotransferases, excluding cytolisis. Although sensitive to distomatosis, γGT and PAL values were inconsistent with the infection rate. The relatively high levels of proteins and albumin eliminate hepatic insufficiency. CONCLUSION: Our results suggest a chronic fasciolosis, confirmed by histology and SEM.

Prevalence of coccidian infection in rabbit farms in North Algeria.

AIM: The aim of the present study was to determine the prevalence and intensity of rabbit coccidiosis (Oryctolagus cuniculus) in North Algeria. MATERIALS AND METHODS: During the study, 40 rabbit farms were investigated. The farms are located in the provinces of Tizi Ouzou, Médéa, and Djelfa which distributed, respectively, into three regions: East Tell Atlas Mountains, Central Tell Atlas Mountains, and High Plateaus. The number of oocyst per gram of feces (OPG) was determined by McMaster technique, and the Eimeria species were identified using morphological criteria. RESULTS: In the farms investigated, the prevalence of coccidian infection was estimated to 90% (80.7-99.3%) in rabbits after weaning. The classification of the farms according to their parasite load allowed us to show that 37.5% of the prospective farms have an oocyst excretion between 104 and 5×104 oocysts per gram and 22.5% excrete >5×104 oocysts per gram. Excretion levels by region show that the region of East Tel Atlas Mountains ranks first with 79% of farms with a parasitic load >104 coccidians compared to the regions of Central Tel Atlas Mountains and High Plateaus. In total, eight species of Eimeria were identified from oocyst-positive samples. Mixed infections with four Eimeria species were common. E. magna is the dominant species in comparison with E. media and E. irresidua with respective frequencies of 42.5% and 17.6% and 14.9% (p<0.001). Our results showed that the farms using anticoccidial drugs for their rabbits were low (25%) and the percentage of farms with poor hygienic conditions was 65%. There was a significant association between increased oocysts excretion and control measures of coccidian infection. CONCLUSION: The study revealed an overall prevalence of 90% in the three Algerian regions. A strong association was observed between Eimeria infection and hygienic status and preventional chemotherapy.

Prevalence of bovine theileriosis in North Central region of Algeria by real-time polymerase chain reaction with a note on its distribution.

To determine the presence and distribution of bovine theileriosis in the North Central region of Algeria, 358 DNA samples and 359 blood smears were analyzed from nine provinces. Theileria DNA extracted from cattle blood was amplified by fluorescence resonance energy transfer polymerase chain reaction (FRET-PCR). Blood smears were examined for Theileria piroplasms by microscopical examination (ME) of Giemsa-stained slides. While microscopical identification revealed only 42 animals being infected with Theileria piroplasms, PCR-positive amplification using Theileria genus-specific primers was obtained from 132 Theileria spp. (P < 0.0001). Among the 132 positives, 108 animals (81.8 %) were found positive of Theileria annulata, while 24 (18.2 %) were found positive for Theileria sp. (P < 0.0001). However, melting curve analysis of these latter samples revealed the presence of two different peaks, 51.5 ± 0.5 °C corresponding to Theileria sp1 and 52.5 ± 0.5 °C for Theileria sp2. Cloning and sequencing of Theileria sp1 and Theileria sp2 using the Cox primers indicated that these species are very closely related to Theileria buffeli. There is a highly significant difference in the distribution of theileriosis between different provinces (P < 0.0001). This disparity between provinces is probably due to differences in tick contact, influenced by the subhumid bioclimatic gradient and differences in agricultural land use.

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.