Seroprevalence Assessment and Risk Factor Analysis of Toxoplasma gondii Infection in Goats from Northeastern Algeria.

T. gondii is the causal agent of toxoplasmosis, a worldwide zoonotic disease relevant in human and veterinary medicine. In Algeria, few reports focused on the presence and circulation of this parasite in the local goat population. The aim of the survey was to evaluate toxoplasmosis seroprevalence and associated risk factors. Sera from 460 goats reared on 72 farms in northeastern Algeria were collected and tested for IgG antibodies to T. gondii by an indirect ELISA. To identify risk factors, a linear regression analysis of the variables was performed. Anti-T. gondii antibodies were found in 94.44% (68/72; 95% CI: 73.34-119.73) of goat farms and in 53.26% (245/460; 95% CI: 46.80-60.36) at the individual level. The multivariable analysis showed that seasonal pasture (OR = 3.804; 95% CI: 3.321-4.358; p = 0.003), presence of water source in pasture area (OR = 4.844; 95% CI: 1.942-7.789; p = 0.0004), use of anthelminthics (OR = 2.640; 95% CI: 1.592-3.146; p = 0.036), number of cats, hygiene, proportion of abortions, number of abortions in the last year, year of sampling, region, and season were the variables significantly associated with T. gondii seropositivity. Abortions in goat herds seem to be related to T. gondii exposure, thus it is crucial to undertake measures and strategies to reduce, control, and prevent toxoplasmosis infection in goats, and thereby in humans, from Algeria.

Molecular characterization of Echinococcus granulosus sensu lato genotypes in dromedary camels from extreme Sahara of Algeria based on analysis of nad2 and nad5 genetic markers.

Cystic echinococcosis is parasitic disease caused by the metacestodes belonging to the Echinococcus granulosus sensu lato (s.l.) species complex. Cystic echinococcosis is of considerable economic and public health importance. It is endemic in both livestock and humans in North African countries, including Algeria. The present study aimed to characterize E. granulosus s.l. genotypes in dromedary camels (Camelus dromedarius) from the extreme Sahara of Algeria, using recently developed mitochondrial genetic markers (NADH dehydrogenase subunit 2 and NADH dehydrogenase subunit 5) for reliable identification of different genotypes. A total of 75 Echinococcus cysts were collected from 49 dromedary camels, including 65 and 10 cysts from 45 and four camels originating from two slaughterhouses of Tindouf and Illizi provinces, respectively. E. granulosus sensu stricto (s.s.) G1 and G3 were identified in camels from both areas based on nad5 (649 bp) gene sequences, whereas E. granulosus s.l. G6 was identified in camels from Tindouf region based on concatenated nad5 and nad2 gene sequences (total 1336 bp). Identified samples clustered into 11 different haplotypes (ALG1-ALG11), including four haplotypes (ALG8-ALG11) for E. granulosus s.s. G1, one haplotype (ALG7) for E. granulosus s.s. G3, and six haplotypes (ALG1-ALG6) for E. granulosus s.l. G6. The present study provides valuable molecular data, including genotyping and haplotypic variability, on E. granulosus s.l. in dromedary camels from two regions in the extreme Sahara of Algeria. Future characterization of the G1, G3, and G6 samples based on sequencing of complete mitochondrial genomes would be of considerable significance for a more comprehensive understanding of molecular epidemiology of CE in Algeria.

First Epidemiological Report on the Prevalence and Associated Risk Factors of Cryptosporidium spp. in Farmed Marine and Wild Freshwater Fish in Central and Eastern of Algeria.

PURPOSE: The present study aimed to estimate the prevalence and molecular characterization of Cryptosporidium spp. in six different fish species both from marine and freshwater environments. METHODS: During a period of 2 years (2018-2020), a total of 415 fecal samples and 565 intestinal scrapings were collected in seven provinces from the central and eastern Algeria. From those, 860 fish belonged to six different species, two of which are cultured marine and four are wild freshwater fish. All samples were screened for Cryptosporidium spp. presence using molecular techniques. Nested PCR approach was performed to amplify partial sequences of the small subunit ribosomal RNA (SSU rRNA) and 60-kDa glycoprotein (GP60) genes for Cryptosporidium genotyping and subtyping. Detailed statistical analysis was performed to assess the prevalence variation of Cryptosporidium infection according to different risk factors. RESULTS: Nested PCR analysis of SSU gene revealed 173 Cryptosporidium positive fish, giving an overall prevalence of 20.11% (17.5-23.0). Cryptosporidium spp. was detected in 8.93% (42/470) of cultured marine fish and 33.58% (131/390) of wild freshwater fish. Overall, the prevalence was affected by all studied risk factors, except the gender. Molecular characterization and subtyping of Cryptosporidium isolates showed occurrence of IIaA16G2R1 and IIaA17G2R1 subtypes of C. parvum in the fish species Sparus aurata. CONCLUSION: The present study provides the first epidemiological data on the prevalence and associated risk factors of Cryptosporidium spp. in farmed marine and wild freshwater fish and the first molecular data on the occurrence of zoonotic C. parvum in fish from North Africa (Algeria).

Prevalence and risk factors of intestinal protozoan infection among symptomatic and asymptomatic populations in rural and urban areas of southern Algeria.

BACKGROUND: Intestinal parasitic infections are amongst the most common infections worldwide and have been identified as one of the most significant causes of morbidity and mortality among disadvantaged populations. This comparative cross-sectional study was conducted to assess the prevalence of intestinal protozoan infections and to identify the significant risk factors associated with intestinal parasitic infections in Laghouat province, Southern Algeria. METHODS: A comparative cross-sectional study was conducted, involving 623 symptomatic and 1654 asymptomatic subjects. Structured questionnaires were used to identify environmental, socio demographic and behavioral factors. Stool specimens were collected and examined using direct wet mount, formalin-ether concentration, xenic in vitro culture and staining methods. RESULTS: A highly significant difference of prevalence was found between symptomatic (82.3%) and asymptomatic subjects (14.9%), with the majority attributable to protozoan infection. The most common species in the symptomatic subjects were Blastocystis spp. (43.8%), E. histolytica/dispar (25.4%) and Giardia intestinalis (14.6%) and more rarely Enterobius vermicularis (02.1%), Teania spp. (0.6%) and Trichuris trichiura (0.2%), while in asymptomatic population Blastocystis spp. (8%), Entamoeba coli (3.3%) and Entamoeba histolytica/dispar (2.5%) were the most common parasites detected with no case of helminth infection. Multivariate log-linear analysis showed that contact with animals was the main risk factor for transmission of these protozoa in both populations. Furthermore, living in rural areas was significantly associated with combined protozoan infection in the asymptomatic population, whereas, in the symptomatic population an increasing trend of protozoan infection was detected in the hot season. In addition, Blastocystis spp. and G. intestinalis infection were found to be associated with host sex and contact with animals across the study period. CONCLUSIONS: Based on these results, several strategies are recommended in order to effectively reduce these infections including good animal husbandry practices, health education focused on good personal hygiene practices and adequate sanitation.

First report on occurrence of zoonotic helminth Toxocara canis, Toxascaris leonina and Ancylostoma caninum in domestic dogs from province of Djelfa, Algeria.

A total of 85 faecal samples of domestic dogs were collected from six localities, including Birine, Sidi Ladjel, Dar Chioukh, Aïn Maâbed, Aïn El-Ibel and Djelfa city from province of Djelfa, Algeria. Samples were tested for presence of Toxocara canis, Toxascaris leonina and Ancylostoma caninum eggs by formol-ether concentration and flotation techniques. Microscopic examination revealed an infection rate of 9.4%, 15.3% and 1.15% for T. canis, T. leonina and A. caninum, respectively. Males were more frequently infected with T. leonina as compared to females, while, no difference was observed regarding T. canis and A. caninum. Prevalence of T. canis and T. leonina did not vary significantly with age ranges of dogs. In contrast, A. caninum seems to be more found in dogs aged of 6-12 months. Prevalence of the three species did not vary significantly in dogs without diarrhea and those with diarrhea. Through these preliminary findings, dogs from steppic region of Djelfa can play a potential role in the dissemination of these neglected zoonotic helminths, which may endanger health of peoples, particularly, pastoral community.

Gastrointestinal parasites of dromedary camel (Camelus dromedarius) in Algeria.

AIM: The present study was designed to investigate the prevalence and identification of gastrointestinal parasites in feces samples of dromedary camels (Camelus dromedarius) in Algeria based on microscopic examination. MATERIALS AND METHODS: A total of 717 fresh fecal samples obtained from 28 farms at Steppe and Northern Sahara regions of Algeria were processed for microscopic examination after concentration by formalin-ether sedimentation and flotation techniques. In addition, microscopic examination of Cryptosporidium spp. was done by modified Ziehl-Neelsen staining and Lugol staining procedure was used for the detection of Giardia cysts. RESULTS: Microscopic examination indicated an infection rate of gastrointestinal parasites of 48.26% (346/717). Protozoan infections were recorded at 17.02% (122/717), whereas helminth infections were recorded at 23.71% (170/717). In addition, mixed infection (protozoans and helminths) was seen at 7.53% (54/717). No correlation was found between infection and age of the animals, nor the consistency of the stool samples; in addition, neither influence of sex nor breed of camels was observed. Eighteen genera of gastrointestinal parasites were revealed; including four genera of protozoa, 12 Nematoda, one Cestoda, and one Trematoda. Strongyloides spp. and Eimeria spp. showed the highest rate of parasitism, while Cooperia spp. was observed with the lowest prevalence. Cryptosporidium spp. was detected in 13 among 717 examined samples (1.81%). CONCLUSION: The parasite fauna infecting the gastrointestinal tract of the Algerian dromedary is much diversified. The detected parasites in camels are similar to counterparts in other ruminants, posing serious challenge to animal farming. Future studies should be carried out to better understand the epidemiology of these parasitic diseases and their economic and public health impact.

Occurrence and molecular characterization of Giardia duodenalis in lambs in Djelfa, the central steppe of Algeria.

Little is known of the prevalence and genetic identity of Giardia duodenalis in sheep in Algeria. The present study aimed at characterizing G. duodenalis in lambs up to 6 months of age in Djelfa, Algeria. A total of 346 fecal specimens were collected from 28 farms and screened for G. duodenalis cysts by zinc sulfate flotation microscopy, and positive specimens were confirmed using a direct immunofluorescence assay. Microscopy-positive specimens were analyzed by PCR and sequence analysis of the triosephosphate isomerase and glutamate dehydrogenase genes to determine G. duodenalis assemblages. Coprological examination indicated that the overall infection rate was 7.0% (24/346). Lambs under 3 months of age had higher infection rate (18/197, 9.0%) than older (6/149, 4.0%) animals, and animals with diarrhea (7/44, 16.0%) had higher infection rate than animals without diarrhea (17/302, 5.6%). PCR sequence analyses of the 15 G. duodenalis isolates revealed the presence of assemblages A in 6 isolates, assemblage E in 7 isolates, and both in 2 isolates. Assemblage A was only found in pre-weaned lambs with diarrhea, while assemblage E was mostly found in post-weaned lambs without diarrhea. The assemblage E isolates from sheep were genetically related to those from cattle in Algeria, while assemblage A isolates were from a well-known subtype prevalent in humans. Data generated from the study improve our understanding of the transmission of G. duodenalis in Algeria.

Zoonotic Cryptosporidium species and subtypes in lambs and goat kids in Algeria.

BACKGROUND: Little is known on the occurrence and identity of Cryptosporidium species in sheep and goats in Algeria. This study aimed at investigating the occurrence of Cryptosporidium species in lambs and goat kids younger than 4 weeks. METHODS: A total of 154 fecal samples (62 from lambs and 92 from kid goats) were collected from 13 sheep flocks in Médea, Algeria and 18 goat flocks across Algiers and Boumerdes. They were screened for Cryptosporidium spp. by nested-PCR analysis of a fragment of the small subunit (SSU) rRNA gene, followed by restriction fragment length polymorphism and sequence analyses to determine the Cryptosporidium species present. Cryptosporidium parvum and C. ubiquitum were further subtyped by sequence analysis of the 60 kDa glycoprotein gene. RESULTS: Cryptosporidium spp. were detected in 17 fecal samples (11.0%): 9 from lambs (14.5%) and 8 from goat kids (8.7%). The species identified included C. parvum in 3 lambs, C. xiaoi in 6 lambs and 6 goat kids, and C. ubiquitum in 2 goat kids. Cryptosporidium infections were detected mostly in animals during the first two weeks of life (7/8 for goat kids and 7/9 for lambs) and in association with diarrhea occurrence (7/17 or 41.2% goat kids and 7/10 or 70.0% lambs with diarrhea were positive for Cryptosporidium spp.). Subtyping of C. parvum and C. ubiquitum isolates identified the zoonotic IIaA13G2R1 and XIIa subtype families, respectively. Minor differences in the SSU rRNA gene sequences were observed between C. xiaoi from sheep and goats. CONCLUSIONS: Results of this study indicate that three Cryptosporidium species occur in lambs and goat kids in Algeria, including zoonotic C. parvum and C. ubiquitum. They are associated with the occurrence of neonatal diarrhea.

Genotypes and subtypes of Cryptosporidium spp. in diarrheic lambs and goat kids in northern Greece.

Inconsistent data exist on the distribution of zoonotic Cryptosporidium species and subtypes in sheep and goats in European countries, and few such data are available from Greece. In this study, 280 fecal specimens were collected from 132 diarrheic lambs and 148 diarrheic goat kids aged 4 to 15 days on 15 farms in northern Greece, and examined for Cryptosporidium spp. using microscopy of Ziehl-Neelsen-stained fecal smears. Cryptosporidium spp. in 80 microscopy-positive fecal specimens (39 from lambs and 41 from goat kids) were genotyped by PCR-RFLP analysis of the small subunit rRNA gene and subtyped by sequence analysis the 60 kDa glycoprotein gene. Among the 33 specimens successfully genotyped, C. parvum was found in 32 and C. xiaoi in one. Seven subtypes belonging to two subtype families (IIa and IId) were identified among the 29 C. parvum specimens successfully subtyped, including IIaA14G2R1 (1/29), IIaA15G2R1 (6/29), IIaA20G1R1 (7/29), IIdA14G2 (1/29), IIdA15G1 (9/29), IIdA16G1 (3/29), and IIdA23G1 (2/29). Lambs were more commonly infected with C. parvum IIa subtypes, whereas goat kids were more with IId subtypes. The results illustrate that C. parvum is prevalent in diarrheic lambs and goat kids in northern Greece and these animals could potentially play a role in epidemiology of human cryptosporidiosis.

Divergent Cryptosporidium parvum subtype and Enterocytozoon bieneusi genotypes in dromedary camels in Algeria.

Little information is available on the occurrence of the zoonotic protists Cryptosporidium spp. and none on Enterocytozoon bieneusi in camels. This preliminary study was conducted to examine the identity of Cryptosporidium subtypes and E. bieneusi genotypes in dromedary camels in Algeria. A total of 39 fecal specimens were collected from young camels. PCR-sequence analysis of the small subunit rRNA was used to detect and genotype Cryptosporidium spp. Cryptosporidium parvum present was further subtyped by sequence analysis of the 60 kDa glycoprotein gene. PCR-sequence analysis of the ribosomal internal transcribed spacer gene was used to detect and genotype E. bieneusi. Altogether, two and eight of the specimens analyzed were positive for C. parvum and E. bieneusi, respectively. The former was identified as a new subtype that is genetically related to the C. hominis If subtype family, whereas the latter was identified as two related genotypes (Macaque1 and a novel genotype) in the newly assigned E. bieneusi genotype group 8. Although they are not known hosts for C. parvum and E. bieneusi, camels are apparently infected with genetically distinct variants of these pathogens.

Molecular characterization of zoonotic pathogens Cryptosporidium spp., Giardia duodenalis and Enterocytozoon bieneusi in calves in Algeria.

Little is known on the identity and public health potential of Cryptosporidium spp., Giardia duodenalis and Enterocytozoon bieneusi in farm animals in Algeria. In this study, 102 fecal specimens from pre-weaned dairy calves with or without diarrhea were collected from 19 dairy farms located in 6 provinces. PCR-restriction fragment length polymorphism analysis of the small subunit rRNA gene was used to detect and differentiate Cryptosporidium spp., whereas PCR-sequence analysis of the triosephosphate isomerase gene and ribosomal internal transcribed spacer were used to detect and genotype G. duodenalis and E. bieneusi, respectively. Cryptosporidium was found in 14 specimens, among which 7 had C. parvum, 4 had C. bovis, and 3 had mixed infection of C. parvum and C. bovis or C. bovis and C. andersoni. Subtyping of C. parvum by PCR-sequence analysis of the 60kDa glycoprotein gene identified two zoonotic subtypes IIaA16G2R1 and IIaA17G3R1. G. duodenalis was found in 28 specimens, with 6 having the host-specific assemblage E, 14 having the zoonotic assemblage A (all belonging to A2 subtype), and 8 having mixed assemblages. Six known genotypes of E. bieneusi belonging to Group 2, including I, J, BEB3, BEB4, BEB6 and PtEb XI, were identified in 11 specimens. Diarrhea was mostly associated with the occurrence of C. parvum. Data from this study suggest that human-pathogenic C. parvum subtypes and G. duodenalis and E. bieneusi genotypes are common on dairy farms in Algeria.

Common occurrence of zoonotic pathogen Cryptosporidium meleagridis in broiler chickens and turkeys in Algeria.

Only a small number of birds have been identified by molecular techniques as having Cryptosporidium meleagridis, the third most important species for human cryptosporidiosis. In this study, using PCR-RFLP analysis of the small subunit (SSU) rRNA gene, we examined the ileum of 90 dead chickens from 23 farms and 57 dead turkeys from 16 farms in Algeria for Cryptosporidium spp. C. meleagridis-positive specimens were subtyped by sequence analysis of the 60 kDa glycoprotein gene. Cryptosporidium infection rates were 34% and 44% in chickens and turkeys, respectively, with all positive turkeys (25) and most positive chickens (26/31) having C. meleagridis. All C. meleagridis specimens belonged to a new subtype family. The frequent occurrence of C. meleagridis in chickens and turkeys illustrates the potential for zoonotic transmission of cryptosporidiosis in Algeria.