Epidemiological update of cystic echinococcosis in livestock and assessment of practices related to its control in the Mediterranean area.

Cystic echinococcosis (CE), caused by the tapeworm Echinococcus granulosus, is a zoonotic parasitic disease that still represents a serious threat to human and animal health worldwide. The Mediterranean basin is recognized as one of the major hotspots of CE due to several factors, including the presence of diverse intermediate host species as well as socio-economic and cultural conditions of local communities. This study aims to take a closer look at epidemiological data on CE in the Mediterranean area and assess the knowledge attitudes and practices of shepherds towards this disease in four countries (Algeria, Greece, Italy and Tunisia), highly endemic for CE, with the final goal of identifying highly endemic risk areas and practices in use which might potentially allow the persistence of E. granulosus infection in these areas. To update the epidemiological scenario of CE in Mediterranean areas, a comprehensive review of peer-reviewed literature on CE prevalence data published during the 2017-2023 period was carried out and, through a geographical information system (GIS), a map displaying the current CE distribution in the Mediterranean area was generated. In addition, a questionnaire survey was conducted through in-depth interviews of the farmers to collect information on their management system as well as knowledge attitudes and practices towards CE. From the farmer-participatory survey some risky practices emerged including the non-regular deworming of dogs or the use of ineffective drugs or dosing, as well as the provision of uncooked animal viscera to dogs. Finally, lower levels of knowledge and awareness of the disease was observed among farmers from North Africa compared with those of European countries. In conclusion, the results obtained highlight that CE is still a very serious problem in Mediterranean areas and increased efforts are needed to promote awareness among farmers and to turn research results into policy in order to reduce the spread of this disease, according to the One Health perspective.

Comparison of the Scolicidal Activity of Two Leaves Extracts of Myrtus communis from Algeria Against Echinococcus granulosus Sensu Lato Protoscoleces.

PURPOSE: During cystic echinococcosis surgery, the use of scolicidal agents such as hypertonic saline (20%) aims to reduce the risk of infection recurrence, but most of the used agents are associated with undesirable side effects. Therefore, the use of natural scolicidal agents such as medicinal plant extracts could reduce these medical issues. The present study aimed to compare in vitro the scolicidal activity between two extracts of the medicinal plant Myrtus communis from Algeria against Echinococcus granulosus sensu lato protoscoleces. METHODS: The ethanolic and aqueous extraction of plant leaves was performed. Phytochemical analysis by gas chromatography-tandem mass spectrometry (GC-MS/MS), determination of total phenolic and flavonoid contents, and in vitro antioxidant activity by DPPH were evaluated for both extracts. Finally, the in vitro scolicidal activity was tested by different concentrations. The viability was evaluated by the eosin exclusion test. RESULTS: The phytochemical analysis revealed 28 components for the ethanolic extract and 44 components for the aqueous extract. The major components were 2'-hydroxy-5'-methoxyacetophenone and 4-amino-2-methylphenol, respectively. The total phenolic and flavonoid contents were 45.9 ± 0.085 mg of gallic acid equivalent per g of extract (GAE/g E) and 16.5 ± 0.004 mg of quercetin equivalent per g (QE/g E) for the ethanolic extract, and 36.5 ± 0.016 mg GAE/g E and 18.2 ± 0.023 mg QE/g E for the aqueous extract, respectively. Furthermore, ethanolic and aqueous extracts of M. communis gave a value of IC50 = 0.009 ± 0.0004 mg/ml and IC50 = 0.012 ± 0.0003 mg/ml for the antioxidant activity, respectively. The in vitro scolicidal activity with concentrations of 50, 75, 100, and 150 mg/ml was tested for 5, 10, 15, and 30 min, and 5, 10, 15, 30, 60, 90, and 120 min for ethanolic and aqueous extracts, respectively. The mortality rate of protoscoleces at concentrations of 100 and 150 mg/ml was 98.8 and 100%, respectively, after 5 min of exposure to the ethanolic extract, while this rate was 100% at the same concentrations only after 60 min of exposure to the aqueous extract. CONCLUSIONS: The ethanolic extract showed a stronger scolicidal activity against E. granulosus s.l protoscoleces than the aqueous extract. In the future, other investigations are necessary to elucidate the mechanism of action and the possible toxicity on human cells. Moreover, experimental animal studies are required to investigate the efficacy of different extracts of this plant and its components as natural anti-parasitic alternatives for the treatment of human cystic echinococcosis.

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.

An innovative strategy for deworming dogs in Mediterranean areas highly endemic for cystic echinococcosis.

BACKGROUND: Cystic echinococcosis (CE), caused by the larval stage of Echinococcus granulosus sensu lato, is a zoonotic parasitic disease of economic and public health importance worldwide, especially in the Mediterranean area. Canids are the main definitive hosts of the adult cestode contaminating the environment with parasite eggs released with feces. In rural and peri-urban areas, the risk of transmission to livestock as well as humans is high because of the free-roaming behavior of owned/not owned dogs. Collecting data on animal movements and behavior using GPS dataloggers could be a milestone to contain the spread of this parasitosis. Thus, this study aims to develop a comprehensive control strategy, focused on deworming a dog population in a pilot area of southern Italy (Campania region) highly endemic for CE. METHODS: Accordingly, five sheep farms, tested to be positive for CE, were selected. In each sheep farm, all shepherd dogs present were treated every 2 months with praziquantel. Furthermore, 15 GPS dataloggers were applied to sheep and dogs, and their movements were tracked for 1 month; the distances that they traveled and their respective home ranges were determined using minimum convex polygon (MCP) analysis with a convex hull geometry as output. RESULTS: The results showed that the mean daily walking distances traveled by sheep and dogs did not significantly differ. Over 90% of the point locations collected by GPS fell within 1500 mt of the farm, and the longest distances were traveled between 10:00 and 17:00. In all the sheep farms monitored, the area traversed by the animals during their daily activities showed an extension of < 250 hectares. Based on the home range of the animals, the area with the highest risk of access from canids (minimum safe convex polygon) was estimated around the centroid of each farm, and a potential scheme for the delivery of praziquantel-laced baits for the treatment of not owned dogs gravitating around the grazing area was designed. CONCLUSIONS: This study documents the usefulness of geospatial technology in supporting parasite control strategies to reduce disease transmission.

German Ixodes inopinatus samples may not actually represent this tick species.

Ticks are important vectors of human and animal pathogens, but many questions remain unanswered regarding their taxonomy. Molecular sequencing methods have allowed research to start understanding the evolutionary history of even closely related tick species. Ixodes inopinatus is considered a sister species and highly similar to Ixodes ricinus, an important vector of many tick-borne pathogens in Europe, but identification between these species remains ambiguous with disagreement on the geographic extent of I. inopinatus. In 2018-2019, 1583 ticks were collected from breeding great tits (Parus major) in southern Germany, of which 45 were later morphologically identified as I. inopinatus. We aimed to confirm morphological identification using molecular tools. Utilizing two genetic markers (16S rRNA, TROSPA) and whole genome sequencing of specific ticks (n = 8), we were able to determine that German samples, morphologically identified as I. inopinatus, genetically represent I. ricinus regardless of previous morphological identification, and most likely are not I. ricinus/I. inopinatus hybrids. Further, our results showed that the entire mitochondrial genome, let alone singular mitochondrial genes (i.e., 16S), is unable to distinguish between I. ricinus and I. inopinatus. Our results suggest that I. inopinatus is geographically isolated as a species (northern Africa and potentially southern Spain and Portugal) and brings into question whether I. inopinatus exists in central Europe. Our results highlight the probable existence of I. inopinatus and the power of utilizing genomic data in answering questions regarding tick taxonomy.

Molecular characterization and phylogenetic analysis of Argas persicus (Oken, 1818) (Acari: Argasidae) from domestic birds in eastern Algeria.

Argas persicus (the fowl tick) is a species of soft tick commonly associated with poultry farms. It has a wide geographic distribution and colonizes different climate regions. Morphological identification of A. persicus has been reported worldwide, but genetic data regarding its molecular characterization is limited. The present study provides data for morphological identification and genetic characterization of A. persicus collected from domestic birds in traditional farms from east Algeria (Setif region). Additionally, A. persicus samples originating from Gansu province in China were included for comparative molecular study. In total, 1518 ticks collected from 30 infested farms were examined and morphologically identified as A. persicus. Furthermore, the 14 tick samples obtained from China were morphologically identified as A. persicus. Molecular analysis of 30 ticks from Algeria (one tick from each infested farm) and the 14 Chinese samples based on PCR, sequencing, and phylogenetic analysis of three mitochondrial genetic markers (16S rRNA, 12S rRNA, and cox1) confirmed morphological results where all samples belonged to the A. persicus group. However, phylogenetic analysis showed that all Algerian samples and two Chinese samples belong to A. persicus sensu stricto (s.s.), while the remaining Chinese samples represented A. persicus sensu lato (s.l.) (divergent lineage). The present study confirms the occurrence of A. persicus s.s. both in Algeria and China, as well as provides novel molecular data for a distinct Chinese lineage of A. persicus.

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.

Molecular detection of tick-borne pathogens in Rhipicephalus sanguineus sensu stricto collected from dogs in the steppe and high plateau regions of Algeria.

Epidemiology and distributions of canine tick-borne diseases as well as their veterinary and zoonotic significance are poorly understood in Algeria. The present study describes a molecular investigation of important tick-borne pathogens in Rhipicephalus sanguineus sensu stricto collected from domestic dogs in steppe and high plateau areas of central and eastern Algeria. In total, 1,043 ticks were collected from 147 dogs, including 756 ticks from 124 dogs in the steppe region of Djelfa and 287 ticks from 23 dogs in the high plateau area of Bordj Bou Arreridj. Ticks were divided into 384 pools (309 pools from Djelfa and 75 pools from Bordj Bou Arreridj) and tested for genomic materials of Crimean-Congo hemorrhagic fever virus (CCHFV) as well as DNA for Anaplasma phagocytophilum, Anaplasma platys, Ehrlichia canis, Rickettsia spp., Babesia spp., and Hepatozoon spp. using PCR, sequencing and phylogenetic analysis. Hepatozoon spp. was most prevalent, with 160 positive pools (41.7%), and 12 of these were sequenced and identified as Hepatozoon canis. Babesia spp. was detected in 50 samples (13.0%), of which 11 were sequenced and identified as Babesia vogeli. A. platys and E. canis were detected in 92 (24.0%) and 15 (3.9%) of tested samples, respectively. Rickettsia spp. were detected in 24 (6.3%) samples, including 11 samples identified as R. massiliae, 6 samples identified as R. conorii conorii, and 7 samples could not be identified to species level. All 384 pools tested negative for CCHFV and A. phagocytophilum. In addition to detection of R. conorii conorii, R. massiliae, and E. canis, the present study provides the first molecular data for occurrence of A. platys, B. vogeli, and H. canis in Rh. sanguineus s.s. infesting dogs in Algeria. Further large scale studies should be conducted to better understand the epidemiology, distributions, and importance of canine tick-borne pathogens 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).

Epidemiological profile and fertility assessment of hydatid cysts surgically removed from patients in Djelfa province, Algeria.

A survey on human cystic echinococcosis was carried out in two public health establishments including the state hospital and one private medical clinic in Djelfa province (Algeria) to report the epidemiological profile of surgical cases and to examine the fertility of collected cysts. Total 18 hydatid cysts from 11 patients of different ages living both in rural and urban areas were obtained. Liver localization was noted in 73% of cases compared to lung localization (27%). Microscopic examination showed a fertility rate of 94.45% against 5.55% of sterile cysts.

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.

Morphological anomalies in Rhipicephalus sanguineus s.s. (Acari: Ixodidae) collected from dogs in steppe and high plateaus regions, Algeria.

The present report describes morphological anomalies in Rhipicephalus sanguineus s.s. collected from dogs between February 2017 and May 2019 in the steppic region of Djelfa and high plateau area of Bordj-Bou-Arreridj in Algeria. Out of 1043 ticks from 147 infested dogs, four adult ticks (0.38%), collected from four dogs, exhibited local morphological abnormalities. One male displayed ectromely of leg I left, and the other three specimens exhibited multiple associated anomalies in idiosomas and exoskeletons, festoons, stigmas (spiracles), as well as in adanal and accessory plates. This report provides additional exemplars of morphological abnormalities in tick populations of Africa, particularly North Africa.

Prevalence and Clinical Manifestations of Giardia intestinalis and Other Intestinal Parasites in Children and Adults in Algeria.

Giardia intestinalis is one of the most common causes of parasite-induced diarrhea, abdominal pain, flatulence, and malabsorption. Yet, data on the epidemiology of G. intestinalis infections in North Africa are limited. The purpose of this study was to carry out a retrospective survey on the level of intestinal parasitism with a particular emphasis on G. intestinalis in children and adults in Algiers, Algeria. A total of 2,054 individuals from outpatient clinics or hospitalized at Beni-Messous University Hospital of Algiers undergoing stool microscopy for ova and parasites were included. The overall parasite infection rate was 28%. In the 567 parasite-positive samples, Blastocystis was found most frequently (57.3%), followed in frequency by Endolimax nana (41.0%), Entamoeba histolytica/dispar (19.6%), G. intestinalis (17.1%), Entamoeba coli (13.9%), Chilomastix mesnili (1.0%), Iodamoeba bütschlii (0.7%), Entamoeba hartmanni (0.5%), and Cryptosporidium spp. (0.2%). Intestinal parasites were generally more common in adults than in children, except for Giardia, which was more common in children (P = 0.0001). Giardia infection was independent of gender (P = 0.94). Compared with other intestinal parasitic infections, clinical manifestations, such as abdominal pain (P = 0.28) and diarrhea (P = 0.82), were found not to be significantly linked to Giardia infection. In conclusion, G. intestinalis is common in individuals referred to the University Hospital of Beni-Messous with digestive symptoms, particularly so in children. However, in our study, intestinal symptoms appeared not to be more linked to Giardia than to other intestinal parasites.

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.

Mitochondrial 16S rRNA gene analysis reveals occurrence of Rhipicephalus sanguineus sensu stricto from steppe and high plateaus regions, Algeria.

The present study aims to perform comparative genetic characterization for R. sanguineus s.l. infesting Algerian dogs living in steppe and high plateau regions based on mitochondrial 16S rRNA gene sequencing and phylogenetic analysis. In total, 1043 ticks were collected from 147 infested dogs, including 756 ticks from 124 dogs in Djelfa and 287 ticks from 23 dogs in Bordj-Bou-Arreridj. All ticks were morphologically identified as R. sanguineus s.l., and 87 specimens were randomly selected for molecular analysis. Sequences obtained from selected ticks clustered with sequences of R. sanguineus s.l. from France, Germany, Portugal, and Spain, all of which belong to R. sanguineus s.s. The occurrence of R. sanguineus s.s. in Algeria is an interesting result given that most R. sanguineus in Africa cluster among the tropical lineage. This finding should prompt further investigation by exhaustive studies across Algerian territory. Such data can be useful to understand the dispersal route of R. sanguineus s.s. from Europe to North Africa, or whether their occurrence in Algeria is a result of historical biogeography.