New Haplotypes of Blastocystis sp. Identified in Faeces from Various Animal Groups in Algeria.

PURPOSE: Blastocystis sp. is a single-celled, anaerobic, parasitic protozoan commonly found in the intestinal tract of animals and humans globally. Genetic analysis has revealed significant diversity within its species, leading to the identification of at least 40 subtypes (ST1-ST40). This study aimed to identify and differentiate Blastocystis in faeces samples from various animal hosts in Algeria. METHODS: A total of 403 fecal samples, collected from both domestic and zoo animals, were subjected to PCR amplification and sequencing of Blastocystis-specific small subunit ribosomal RNA (SSU-RNA) gene. RESULTS: The overall prevalence of Blastocystis in animals was found to be 38.9%. Through comprehensive phylogenetic and phylogeographic analyses, we identified four distinct subtypes (ST1 in both domestic and zoo animals, and ST3, ST4, and ST5 exclusively in zoo animals), encompassing nine different haplotypes, including five that appear original to Algeria. CONCLUSION: This study represents the first epidemiological molecular investigation of Blastocystis sp. in animals in Algeria.

Microscopic and molecular prevalence and associated risk factors with Toxocara and Blastocystis infection in dogs and cats in Mitidja, Algeria.

Domestic dogs and cats can serve as a source of environmental contamination with Toxocara spp. and Blastocystis spp., and this represents a neglected public and veterinary health problem. We assessed the microscopic and molecular prevalence of these species in a locality in Algeria and identified the associated risk factors. The faeces of 225 dogs and 78 cats were collected in Mitidja between March and July 2022. The samples were analysed by coproscopy and by polymerase chain reaction (PCR) targeting the Internal Transcribed Spacer 2 (ITS2) and Small Subunit Ribosomal (SSU-RNA) of T. canis and Blastocystis spp. respectively. The overall microscopic prevalence of Toxocara spp. in dogs and cats was 9.78 ± 1.98% and 12.82 ± 7.42%, respectively. The rate of Blastocystis spp. was 15.11 ± 2.39% and 15.38 ± 4.08% in dogs and cats, respectively while the molecular prevalence of T. canis in dogs was 4.89 ± 1.44% and in cats 1.28 ± 1.27%; the prevalence of Blastocystis spp. was 41.78 ± 3.29% and 34.62 ± 5.39% in dogs and cats, respectively. Phylogenetic and phylogeographic analyses identified the presence of the H1 subtype of T. canis in dogs, and the ST1 subtype of Blastocystis in dogs and cats. Dogs with clinical signs were more likely to be infected with T. canis (OR 6.039, P < 0.05) than healthy dogs. This study demonstrates that dogs and cats are carriers of Toxocara spp. and Blastocystis spp. and are therefore a source of environmental contamination. Veterinarians and human health professionals should work together to implement control strategies as part of a "One Health" approach to improving animal health and reducing the risk of transmission to humans.

Characterisation of field tropical Theileriosis and associated risk factors in two bioclimatic areas of Algeria.

Tropical theileriosis (TT) is a tick-borne disease caused by Theileria annulata and commonly infects cattle in tropical and subtropical regions, including Algeria. It is a significant obstacle to cattle breeding programs established to improve production in Algeria. The present investigation aimed to estimate the current molecular prevalence, risk factors, and genetic characterisation of T. annulata in two bioclimatic areas of Algeria. In a cross-sectional study, 679 blood samples (629 from healthy cattle selected on farms and 50 from diseased cattle identified by veterinarians) were collected from the humid (n = 307+50) and semi-arid (n = 322) areas and screened by blood smear examination followed by polymerase chain reaction targeting cytochrome oxidase subunit 3 (cox III) mitochondrial and the 18S ribosomal RNA (18S rRNA) genes for Theileria spp. Seventy-six positive samples (56 clinically healthy and 20 with clinical signs) for Theileria spp. were confirmed to be T. annulata by the merozoïtes surface antigen-1 (Tams1) gene showing a rate of 8.9 % in clinically healthy and 40.0 % in suspected cattle. Among the 307 bloods samples collected from healthy cattle in the humid area, 25 cattle (8.1 %) were positive for T. annulata. Of the 322 healthy cattle from the semi-arid site, 31 (9.6 %) were carriers of T. annulata DNA. In subclinical population, demographic and environmental parameters analysis indicated that T. annulata infection was higher in adult crossbred cattle raised in the intensive and semi-intensive system (P<0.001). The multiple logistic regression analysis showed that age, breed, farming system, and bioclimatic area are potential risk factors for T. annulata infection in cattle (P<0.05). Multiple alignments of cox III sequences of T. annulata showed high heterogeneity with 25 polymorphic sites (nucleotide diversity π = 0.02402), resulting in two haplotypes with a low genetic diversity index (Hd) of 0.533. The 18S rRNA sequence alignment revealed only one T. annulata genotype with 100 % identity to the strains isolated from cattle and ticks in Mediterranean and Asian countries. Our preliminary results will serve as a basis for further studies on the genetic diversity and molecular epidemiology of T. annulata.

Prevalence and risk factors associated with gastrointestinal parasites of pet dogs in North-Central Algeria.

The prevalence and risk factors associated with gastrointestinal parasites in dogs were conducted in Blida, North-Central Algeria. The study was carried out over 131 clinically healthy dogs, from March to June 2019, by coprological methods. Of the 131 collected dogs, 61.07% (n = 80) were found infected by gastrointestinal parasites. Sixty-four dogs were carriers of a single infection with the following parasites Ancylostoma spp (15.27%), Uncinaria spp (14.50%) Toxocara canis (4.58%), Trichuris vulpis (3.82%), Toxascaris leonina (2.29%), Taenia/Echinococcus spp. (2.29%), Mesocestoides spp (0.76%), Cystoisospora spp. (3.05%) and Neospora caninum-like (2.29%). Sixteen dogs harbored mixed infection. Male (OR = 1.18) German shepherds' dogs were more infected (OR = 1.08) by helminthic parasites (OR = 13.64). The frequency of single infections (OR = 6.86) increased with the animal's age (OR = 1.73-3.46). Identifying hookworms, T. canis, and T. vulpis suggests a continuing risk of contamination of pet dogs as a source of human infection with the zoonotic parasites in Blida.

Cross-sectional survey of cattle haemopathogens in Constantine, Northeast Algeria.

This aim of the present study was to estimate the prevalence of haemopathogens in cattle in Beni Hamidene locality, district of Constantine (Νortheastern Algeria). Between June and October 2014, 169 bovines from 25 farms were included in this survey, 32 (18.9%) among them were suspected of piroplasmosis and/or anaplasmosis. Infection prevalences were estimated by microscopic examination of Giemsa-stained blood smears and blood samples from all included cattle (n = 169). Animals were infected by Theileria annulata (65/169; 38.46%), Anaplasma marginale (22/169; 13%) and Babesia bovis (5/169; 3%). Two co-infection patterns were found: Theileria annulata/Anaplasma marginale (7.69%) and Theileria annulata/Babesia bovis (1.18%). Only one farm had no cattle infected by any of the haemopathogens. There was a signification difference of T. annulata infection prevalence according to age category (p =.04). These results emphasised mainly the presence of bovine tropical theileriosis in northeastern, Beni Hamidene locality, province of Constantine, Algeria.

Bovine piroplasmosis-anaplasmosis and clinical signs of tropical theileriosis in the plains of Djurdjura (north Algeria).

The study was conducted during tick activity season over a period of 5 years in the Djurdjura Plains, Algeria. A total of 299 cattle (Holstein, Montbeliard, Fleckvieh and crossbred animals) with clinical signs were included in this study. A total of 171 animals were found positive for at least one pathogen by Giemsa-stained blood smears examination Theileria annulata (136/299, 45.5%), Babesia bovis (14/299, 4.7%), B. bigemina (3/299, 1.0%) and Anaplasma marginale (12/299, 4.0%) were identified. Six animals were co-infected by T. annulata and A. marginale. Although no ticks were collected from diseased animals, clinical signs in cattle were hyperthermia (120/136, 88.3%), gluttony followed by anorexia (113/136, 83.1%), lymph node enlargement (99/136, 72.8%), anaemia (82/136, 60.3%), icterus (58/136, 42.6%) and haemoglobinuria (36/136, 26.5%). Gluttony followed by anorexia was considered highly suggestive of an incubation of tropical theileriosis as shown by a higher receptivity index (IR = 0.89-1). This clinical sign is evident in young Montbeliard and young Holstein males with anaemia (IR = 1) and icterus (IR = 0.78-0.81) which is earlier than haemoglobinuria (IR = 0.51-0.54). The incidence of T. annulata was maximum in July (n = 57), as well as B. bovis (n = 6) and A. marginale (n = 13). These results highlight the preponderance of tropical theileriosis in north-central Algeria, where gluttony followed by anorexia is probably a prodromal symptom during the incubation period of the disease.

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.