Molecular detection and associated risk factors of Anaplasma marginale, A. ovis and A. platys in sheep from Algeria with evidence of the absence of A. phagocytophilum.

Anaplasma species are obligate intracellular rickettsial pathogens that cause significant diseases in animals and humans. Despite their importance, limited information on Anaplasma infections in Algeria has been published thus far. This study aimed to assess the infection rate, characterize Anaplasma species, and identify associated risk factors in selected sheep farms across Oum El Bouaghi region in Algeria. In 2018, we collected 417 blood samples from sheep (Ovis aries) and performed molecular characterization of Anaplasma species infecting these animals. This characterization involved the use of 16S rRNA, msp2, rpoB, and msp5 genes, which were analyzed through nested PCR, qPCR, cPCR, DNA sequencing, and subsequent phylogenetic analysis. Our findings revealed infection rates of 12.7 % for Anaplasma species detected, with Anaplasma ovis at 10.8 %, Anaplasma marginale at 1.7 %, and Anaplasma platys at 0.2 %. Interestingly, all tested animals were found negative for Anaplasma phagocytophilum. Statistical analyses, including the Chi-square test and Fisher exact test, failed to establish any significant relationships (p > 0.05) between A. ovis and A. platys infections and variables such as age, sex, sampling season, and tick infestation level. However, A. marginale infection exhibited a significant association with age (p < 0.05), with a higher incidence observed in lambs (5.2 %) compared to other age groups. Remarkably, this study represents the first molecular detection of A. platys and A. marginale in Algerian sheep. These findings suggest that Algerian sheep may serve as potential reservoirs for these pathogens. This research contributes valuable insights into the prevalence and characteristics of Anaplasma infections in Algerian sheep populations, emphasizing the need for further investigation and enhanced surveillance to better understand and manage these diseases.

The first study on seroprevalence and risk factors of Neospora caninum infection in pregnant local cows from Northeast Algeria.

Background and Aim: Neospora caninum is one of the most common infectious organisms worldwide that causes abortion in cattle. To the best of our knowledge, no previous studies have focused on N. caninum infection in the local Atlas brown cattle from Northeast Algeria. This study aimed to assess the importance of bovine neosporosis for causing abortion in Atlas brown cattle and to identify selected risk factors. Materials and Methods: A case-control study was performed on 60 control farms and 30 case farms. We collected 650 blood samples from 650 pregnant cows from 90 farms in five Algerian provinces; Jijel, Skikda, Annaba, El-Tarf, and Souk-Ahras. Sera samples were analyzed for the presence of antibodies against N. caninum using enzyme-linked immunosorbent assays. Results: The seroprevalence of N. caninum infection in the cows was 36.2% (95% confidence interval [CI]: 32.7-39.8) and in the farms was 81.1% (95% CI: 73.0-89.2). Risk factors found by multivariable logistic regression included: Presence of dogs (odds ratio [OR] 4.7, 95 CI 2.9-7.3); age ≥84 months (OR 4.9, 95 CI 2.8-8.3); Jijel region (OR 2.2, 95 CI 1.1-4.5); white (OR 2.5, 95 CI 1.4-4.4) and gray (OR 2.5, 95 CI 1.4-4.5) coat; moderate (OR 2.30, 95 CI 1.4-3.8) and bad (OR 3.1, 95 CI 1.8-5.3) hygiene; and second (OR 2.5, 95 CI 1.4-4.4); and last (OR 2.3, 95 CI 1.3-4.2) stage of pregnancy. Our case-control study showed no significant association between seropositivity of N. caninum and abortion at the farms level (OR 0.9, 95 CI 0.3-2.7). Similarly, there was no significant association between seropositivity of N. caninum and abortion at the individual level (OR 0.8, 95 CI 0.6-1.2). Conclusion: This is the first study of N. caninum infection in pregnant local cows from Northeast Algeria. The prevalence rate of antibodies against N. caninum was high. Almost all risk factors studied for infection were significantly associated with seroprevalence. Our analysis showed no relation between N. caninum infection and abortion. Consequently, these local cows are resistant to abortion caused by N. caninum.

Effect of Pistacia lentiscus L. Vegetable Oil on Growth Performance and Coccidiosis in Broiler Chickens: In vitro and In vivo Assessment.

This study aims to assess the ability of vegetable oil of Pistacia lentiscus L. (lentiscus oil) in stimulating growth performance of broiler chickens and protecting them against coccidiosis. For this purpose, an in vitro test was first carried out to evaluate the destructive effect of this oil on Eimeria spp oocysts. On the other hand, an in vivo study was carried out to evaluate, once again, the capacity of the vegetable oil of Pistacia lentiscus L. in stimulating broilers growth performance and reducing the coccidiosis clinical signs. Thus, day old chicks were randomly divided into four equal groups: (1) uninfected and not-supplemented control (NI NS); (2) uninfected and supplemented (NI S); (3) infected and not supplemented (I NS); (4) infected and supplemented (IS). Each group was divided into three replicates containing each of them two subjects. The experimental groups (2 and 4) are supplemented with lentiscus oil by force-feeding at the rate of 1 mL per day from the 18th day until the end of the experiment. The chicks of the third and the fourth group are inoculated orally with sporulated oocysts (6.5 × 105 oocysts of Eimeria spp) on the day 20 of age. The results showed that lentiscus oil has an anticoccidial dose-dependent effect as shown by oocysts counting and released substances measurement at 273 nm. The growth performance of the (NI S) group was found better with an improvement percentage of 9.14% compared to the control (p < 0.05). Likewise, the weight gain of the (I S) group seems slightly higher than that of the control one (1316 g and 1235 g, respectively) (p < 0.05). In conclusion, the vegetable oil of Pistacia lentiscus L. has shown, not only, a promising growth effect in broiler chickens, but also, it seems to have a protective effect against coccidiosis sequels caused by Eimeria acervulina infection.

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.

Molecular Characterization of Giardia duodenalis in Children and Adults Sampled in Algeria.

The molecular epidemiology of giardiasis in Africa remains unclear. A study was carried out across four hospitals in Algeria. A total of 119 fecal samples from 55 children, 37 adults, and 27 individuals of undetermined age, all scored positive for intestinal parasites by microscopy, and were screened by real-time PCR for Giardia. Molecular characterization of Giardia was performed by assemblage-specific PCR and PCR targeting the triose phosphate isomerase gene (tpi). Of the 119 samples, 80 (67%) were Giardia-positive by real-time PCR. For 48 moderately-highly real-time PCR-positive samples, tpi genotyping assigned 22 samples to Assemblage A and 26 to Assemblage B. Contrary to Assemblage A, Assemblage B exhibited substantial genetic diversity and allelic heterozygosity. Assemblage-specific PCR proved to be specific for discriminating Assemblage A or B but not as sensitive as tpi genotyping. We confirmed that real-time PCR is more sensitive than microscopy for detecting Giardia in stool samples and that robust amplification and sequencing of the tpi gene is feasible when moderate-to-strongly real-time PCR-positive samples are used. This study is one of the few performed in Africa providing genotyping data on Giardia infections in humans. Both assemblages A and B were commonly seen and not associated with specific sociodemographic data.

Seroprevalence of Antibodies Against Anaplasma Phagocytophilum and Borrelia Burgdorferi in Horses (Equus Caballus) from Northern Algeria.

INTRODUCTION: Horses (Equus caballus) are susceptible to tick-borne diseases. Two of them, Lyme borreliosis due to Borrelia burgdorferi and granulocytic anaplasmosis due to Anaplasma phagocytophilum were investigated in Algerian horses. The diseases have been less extensively studied in horses and results pertinent to Algeria have not been published. MATERIAL AND METHODS: Blood samples were obtained from 128 horses. IgG antibodies directed against Anaplasma phagocytophilum and Borrelia burgdorferi were detected by an indirect immunofluorescence antibody test (IFAT) and ELISA. The potential effects of age, gender, breed, and health status on seropositivity were also evaluated. RESULTS: Using IFAT, 28 (21.8%) and 25 (19.5%) animals were positive for B. burgdorferi and A. phagocytophilum, respectively. Using ELISA, 19 (14.8%) and 33 (25.9%) animals were positive for these bacteria. CONCLUSION: The study shows that horses in Algeria are exposed or co-exposed to tick-transmitted zoonotic bacterial species.

Seroprevalence and risk factors of Coxiella burnetii infection in cattle in northeast Algeria.

A cross-sectional study was conducted to determine the seroprevalence and the risk factors associated with C. burnetii infection in cattle in the state of Setif in northeastern Algeria from March 2016 to April 2018. A total of 678 cows animals aged at least 24 months and belonging to 90 herds were randomly selected. A serum sample from each cow was tested for antibodies against C. burnetii using an indirect enzyme-linked immunosorbent assay (ELISA). A structured questionnaire focusing on risk factors for C. burnetii infection was administered to farm owners involved in the study. The individual animal prevalence was 11.36% (77/678) (95%CI 8.97-13.75%), the herd prevalence was 45.56% (41/90) (95%CI 35.27-55.84%), and the within-herd prevalence ranged from 9.09 to 57.14% (mean 23.71%; Q1 11.11%, Q2 or median 20%, Q3 30%). Multivariable logistic regression analysis revealed that contact with other herds (odds ratio (OR) 1.95, 95 CI 1.12-3.42) and purchased animals (OR 2.05, 95 CI 1.14-3.68) was identified as risk factors for seropositivity to C. burnetii, while the use of disinfectants (OR 0.32, 95 CI 0.14-0.72) was identified as protective factor. The results from the present study indicate that C. burnetii is circulating into cattle herds in the region of Setif in Northeastern of Algeria. It is recommended to implement good hygienic practices and measures of biosecurity to reduce the spread of infection between cattle herds and possible exposure of humans.

Seroepidemiological study of the exposure to Toxoplasma gondii among horses in Algeria and analysis of risk factors.

AIM: The aim of this study was to assess the seroprevalence of the Toxoplasma gondii in horses in different parts of Algeria and to determine risk factors for the infection. MATERIALS AND METHODS: A total of 736 blood samples were collected from horses of various breeds, gender, coat colors, and ages. All horses came from various farms, racecourses, and equestrian centers. The seroprevalence was investigated by three different methods: Indirect fluorescent antibody test (IFAT) as reference method, enzyme-linked immunosorbent assay (ELISA), and latex agglutination test (LAT). RESULTS: Out of the 736 sera, 178 (24.18%) were positive for IFAT, 133 (18.07%) for LAT, and 317 (43.07%) for ELISA. It was found that IFAT and LAT were in high agreement (Kappa 0.79), indicating that LAT and IFAT had similar capabilities in the detection of anti-T. gondii antibodies from horse sera. Risk factors analysis based on IFAT results indicated that the habit of the animals was significant risk factors (p≤0.05) for Toxoplasma infection. The seroprevalence was significantly higher in horses living on farms. Moreover, a higher seroprevalence was found in older animals compared to younger ones. Furthermore, the seroprevalence in females was significantly higher than that in males and gelding. Breed, coat color, and water sources are also important factors to influence the seroprevalence of T. gondii. CONCLUSION: The results indicated that T. gondii is present in horses throughout Algeria and thus represents a risk for both human and animal health. These results underline the need to increase the vigilance and the preventive measures against this disease not only to protect the horses but also to limit the spread of the parasite.

Prevalence of The Main Infectious Causes of Abortion in Dairy Cattle in Algeria.

INTRODUCTION: Abortion in cattle is a major source of economic losses for the agriculture sector. It can be due to infectious or non-infectious factors. Among infectious factors, parasites, bacteria, viruses, and fungi can be involved. The present work investigated the prevalence of the main infectious agents of abortion in Algerian cattle. MATERIAL AND METHODS: Altogether 278 non-aborting and 82 aborting cows were analysed. RESULTS: The prevalence ranged from 0% for Tritrichomonas foetus to 15% for Neospora caninum. Additionally, a case-control study was performed to find the association between the presence of the pathogens and the occurrence of abortion in cows. The odds ratios were significant for Neospora caninum, bovine herpes virus 4, BVD virus, Brucella abortus, Salmonella Dublin, Leptospira interrogans serovar Hardjo, and Coxiella burnetii. CONCLUSIONS: The pathogens enumerated here could be major causes of abortion among Algerian cattle.

Prevalence of select vector-borne pathogens in stray and client-owned dogs from Algiers.

Data on the prevalence of vector-borne diseases agents infecting canines in Algeria is currently lacking. The purpose of this study is to assess by serological and molecular methods the prevalence of select arthropod borne-bacterial infections in client-owned and stray dogs. Antibodies to Anaplasma phagocytophilum were the most prevalent at 47.7%, followed by Borrelia burgdorferi s.l. at 37.6%, Ehrlichia canis at 30.0%, Bartonella henselae at 32.4% and Bartonella vinsonii subsp. berkhoffii at 27%. Seroprevalence was statistically significantly higher in stray dogs than those owned by clients. Seropositivity was not associated with health status, except for E. canis. Molecular evaluation indicates that 17.8% of the 213 analyzed dogs were positive for Ehrlichia and Anaplasma with a prevalence of 4.2% for E. canis, 14.1% for Anaplama platys and 0% for A. phagocytophilum. Seven (7.1%) of the tested dogs were positive for Bartonella spp. with two characterized as Bartonella rochalimae, four as B. henselae and one as B.v. subsp. berkhoffii.

Comparison of different serological methods to detect antibodies specific to Neospora caninum in bovine and canine sera.

Neospora caninum is an apicomplexan parasite responsible for paresis in dogs and abortion in cattle worldwide. Dogs serve as a definitive host, while cattle serve as intermediate host. Many different methods have been developed to detect specific antibodies present in cattle and dog serum. In the present study, the dense granule protein NcGRA6 was incorporated in a latex beads agglutination test (LAT), and compared to other serological methods, including enzyme-linked immunosorbent assay, the direct agglutination test, the immunoblot, and the indirect fluorescent antibody test (IFAT). Using the IFAT as the reference method, 100 sera isolated from Algerian cattle and 100 sera isolated from Algerian dogs, both possibly infected with N. caninum, were used to evaluate the LAT. The sensitivity, specificity, and kappa index were calculated for each host species and assay. For dog sera, the sensitivity and the specificity of the LAT was 76% and 100%, respectively. The McNemar test showed that the LAT was not significantly different from IFAT (P > 0.05). For cattle sera, the sensitivity and the specificity of the LAT were 60% and 100%, respectively. The McNemar test indicated that the LAT was significantly different from IFAT (P < 0.01) and that the LAT was only positive for cattle sera with titers of 1:800 or greater, indicating that LAT can be used for cattle in a clinical context. As well, the LAT has the advantage of being easy and rapid to perform compared to the other assays.

Study of the risk factors associated with Neospora caninum seroprevalence in Algerian cattle populations.

Bovine abortions due to Neospora caninum infection were reported worldwide. The situation in Algeria was unknown. For the evaluation of the prevalence of N. caninum and its associated risk factors, 799 cattle belonging to 87 farms of the north and northeast of Algeria were analyzed. The cattle were divided into imported cattle, local cattle and improved cattle corresponding to breeding between imported and local cattle. Sera were examined for the presence of N. caninum antibodies by indirect fluorescence antibody test. The overall seroprevalence for the 87 farms was 52.87% (41.28-62.71%). The overall animal seroprevalence was 19.64% (16.82-22.45%). The seroprevalence of N. caninum in local cattle (34.28%) was significantly higher (p<0.05) than in modern (16.04%) and improved (18.64%) cattle. The risk factors analysis indicated that cattle population, geographical location, dog presence, season, global farm hygiene or the presence of abortion were significantly associated with seroprevalence.

Neospora caninum is associated with abortion in Algerian cattle.

Neospora caninum is a major cause of abortion in cattle worldwide. However, little information is available for Algeria. Accordingly, 799 cattle from 87 farms in the north and northeast of Algeria were enrolled in a seroepidemiological survey. An indirect fluorescence antibody test (IFAT) revealed a seroprevalence of 19.6%. The animals were divided into 3 groups according to their breed: imported European cattle, local breeds, and crossed animals (European × local). Seroprevalences were 16.0%, 34.3%, and 18.6% in groups 1, 2, and 3, respectively. A case control study was performed to investigate the link between global seropositivity to N. caninum and abortion risk in those cattle farms. There was a significant (P < 0.01) association between the seroprevalence against N. caninum and the occurrence of abortion in those farms (odds ratio [OR]  =  12.03). This was also observed at the individual level (OR  =  2.79). The analysis of results according to the breed revealed a significant association between seroprevalence and abortion in groups 1 and 3, but not for group 2, despite the fact that the highest seroprevalence was observed in group 2. Cerebral tissues from 5 aborted fetuses were available for histology and polymerase chain reaction (PCR). One sample was found positive both by histology and by PCR, 2 samples were positive by PCR only, and 2 samples were negative in both tests.

Evaluation of a SRS2 sandwich commercial enzyme-linked immunosorbent assay for the detection of anti-Neospora caninum antibodies in bovine and canine sera.

Neospora caninum is a parasite responsible for abortion in cows and neuromuscular disease in dogs. Serology is the most widely used technique to evaluate the prevalence of N. caninum in different host populations. A sandwich enzyme-linked immunosorbent assay (ELISA) based on the use of an anti-SRS2 monoclonal antibody was evaluated against the indirect fluorescent antibody test for 100 canine sera and against a well-characterized ELISA for 102 bovine sera. In cattle sera, the relative sensitivity and relative specificity were 100%. In dog sera, the relative specificity and relative sensitivity were 94% and 86%, respectively. The kappa value was 1 for bovine sera and 0.77 for canine sera. The seroprevalence was 3.9% in bovine sera and 21-23% in canine sera. The SRS2 sandwich ELISA was considered a valuable tool in both species.

Detection of Neospora caninum in dog organs using real time PCR systems.

Neospora caninum is a parasite responsible for paresis in dogs. The dog can harbour enkysted parasites in several organs. The detection of N. caninum was performed using 3 different real time PCR systems all amplifying the NC5 DNA region. One system was based on Sybrgreen, one on Plexor technology and the last on Taqman probe. Comparison of the three methods indicated that the detection limit was 1 equivalent genome on pure DNA but that this detection limit increased in the presence of foreign DNA using the Sybrgreen and Plexor systems. Therefore, the Taqman system was chosen to detect N. caninum in liver and spleen of naturally infected dogs. The overall prevalence was 32.2%. Comparison between PCR results and serological results using IFAT showed that among the 28 PCR positive dogs only 9 were seropositive and that 8 seropositive dogs were PCR negative. Therefore serology can underestimate the real carriage in dogs. However, PCR methods must be improved in terms of sensitivity and inhibition problems.