Comparison of a Commercial Enzyme-Linked Immunosorbent Assay (ELISA) with the Modified Agglutination Test (MAT) for the Detection of Antibodies against Toxoplasma gondii in a Cohort of Hunting Dogs.

Toxoplasmosis is a zoonotic disease, caused by the protozoan Toxoplasma gondii, affecting most warm-blooded animals. Assessing the seroprevalence of T. gondii in different animal species gives a good estimate of the global circulation of the parasite and the risk for human infections. However, the seroprevalence of T. gondii in dogs is not studied as much as other species, despite their close contact with wildlife and humans in rural or urban environments and evidence that dogs can also be a potential source for human contaminations. A commercial enzyme-inked immunosorbent assay (ELISA) kit to detect anti-T. gondii antibodies in sera of hunting dogs potentially naturally infected, was compared to the modified agglutination test (MAT), used as the reference method. The ELISA presented a sensitivity of 76.5% (CI 95%: 60.0-87.6) and a specificity of 87.7% (CI 95%: 76.7-93.9) and a substantial agreement with the MAT for the detection of canine anti-T. gondii antibodies. Both tests can therefore be used widely for epidemiology studies on T. gondii infections in dogs. With a mean seroprevalence of T. gondii infection in hunting dogs from northern Algeria of 36.8% (CI 95%: 34.9-38.7), this study also highlights the importance of T. gondii seroprevalence studies in companion animals to assess infectious risk for human populations.

Prediction of Trypanosoma evansi infection in dromedaries using artificial neural network (ANN).

Surra is caused by Trypanosoma evansi, a flagellated parasite that affects domestic and wild animals. Surra is a neglected tropical disease causing serious problems to camels breed in Algeria. The aim of our study consists to extract the major risk factors that predict T.evansi infection in dromedaries using artificial neural networks. This investigation was conducted on 115 dromedaries from Ghardaïa district, Southern Algeria. The immune trypanolysis test was used to detect antibodies against T. evansi. Firstly, the gamma test has been used to choose optimal input parameters. The obtained results indicate that the age, gender, breed, clinical manifestations history, herd size, as well as the animal activities were the most predictors of T. evansi infection. Afterward, an artificial neural network method has been performed for modelling the proposed optimal inputs and their accuracy was assessed through seven statistical indicators. The comparative study indicates the effectiveness of the (6-9-1) model trained by the Tansig transfer function. The proposed model has demonstrated a good performance: 0.925 for training data and 0.962 for validation data. Furthermore it could be very useful for the rapid intervention of veterinarians as close as possible to the point-of-care (POC).

Assessment of Trypanosoma evansi prevalence and associated risk factors by immune trypanolysis test in camels from Ghardaïa district, southern Algeria.

Trypanosoma evansi (T. evansi) is a flagellated parasite with worldwide distribution, mainly affecting camels, horses, dogs, buffaloes and wild animals. Trypanosomosis caused by T. evansi, known as surra, is a vector borne disease that affects the health and productivity of camels. The aim of our study was to assess the prevalence of trypanosomosis due to T. evansi in camels by the immune trypanolosis test and to identify associated risk factors. Our cross-sectional study was performed on 161 camels from Ghardaïa district, southern Algeria. A structured questionnaire was used to collect data on individual characteristics (age, gender and breed) husbandry management (herd size and activity of animals) and health conditions (history of abortion and clinical symptoms). The immune trypanolysis test revealed an overall seroprevalence of 9.3% (CI 95%, 5.9-14.9). Possible factors associated with T. evansi infection were analysed by univariate and multivariate logistic regression. The results showed that risk factors for camels were history of symptoms (P = 0.002, OR = 21.91, CI95% = 3.48-169.80), racing activities (P = 0.003, OR = 0.01, CI95% = 0.001-0.18) and small herd size (P = 0.013, OR = 8.22, CI95% = 1.64-49.75). In conclusion, this study showed that T. evansi is endemic in camels of Ghardaïa district. To reduce dissemination of the disease to non-endemic areas, it is recommended to minimise risk factors associated with the infection.

Molecular detection of the B1 gene of Toxoplasma gondii in blood samples of female sheep and goats in Tebessa, northeastern Algeria.

Toxoplasmosis, caused by Toxoplasma gondii, is a parasitic zoonosis of crucial medical and veterinary importance. It is mainly diagnosed by serological methods which are limited by insufficient sensitivity. Therefore, it is necessary to rely on direct detection of the parasite. The present study was aimed for direct detection of the parasite DNA in the blood samples of sheep and goats using PCR targeting the B1 gene. The study was carried out in 20 small ruminant farms between 2016 and 2018 in Tebessa region, part of north-eastern Algeria, and concerned 227 and 91 aborted female sheep and goats respectively. DNA of T. gondii was detected in 35.24 % and 18.68 % blood samples of sheep and goats respectively (p < 0.001). Molecular prevalence was higher in 13-24 month old female sheep (93.33 %) than 1-12 month old female sheep (14.37 %) (p < 0.0001). While, in goats no significant difference was observed in relation to age. Female sheep that aborted between 1-60 days of gestation were found to be more infected (46.41 %) compared to females that aborted between 61-120 days of gestation (12.16 %) (p < 0.001). Whereas, female goats that aborted between 61-120 days of gestation were found to be more infested (30.77 %) compared to females that aborted between 1-60 days of gestation (16.67 %) (p < 0.001). This study revealed that small ruminants are highly infected with T. gondii, which represents a major risk for the consumer in Tebessa. Further studies are needed to improve our knowledge of the different genotypes of T. gondii infecting small ruminant population.

Comparison of serological and molecular tests for detection of Trypanosoma evansi in domestic animals from Ghardaïa district, South Algeria.

Trypanosoma evansi (T. evansi) is a hemoflagellate parasite that affects a broad range of mammalian hosts and that causes a disease called surra. Diagnosis of surra based on clinical symptoms alone is inaccurate. Therefore, a variety of serological and molecular diagnostic tests are used to assist in the detection of T. evansi infections. The aim of this study was to compare the diagnostic performance of four serological tests (CATT/T.evansi, immune trypanolysis, ELISA with purified variant surface glycoprotein RoTat 1.2 and with whole cell lysate) and two molecular PCR tests targeting sequences within the ribosomal genes locus (ITS1 TD PCR and 18S qPCR). Tests were carried out on blood samples from 161 dromedary camels, 93 horses, 129 goats, 168 sheep, 127 bovines and 76 dogs. Latent class analysis was carried out to calculate the sensitivity and specificity of each diagnostic test. Cohen's Kappa test was used to assess the concordance between the different diagnostic tests. Overall positivity rates observed with the serological tests were as follows: 3.1 % with CATT/T.evansi, 4.9 % with ELISA/RoTat 1.2, 3.4 % with ELISA/whole lysate and 2.0 % with immune trypanolysis (TL). Among the 754 samples tested with the molecular tests, 1.7 % were positive with 18S qPCR and 1.3 % with ITS1 TD PCR. Cohen's Kappa test showed agreement ranging from fair to substantial (k = 0.2-0.8) between serological diagnostic tests. However, it showed a perfect agreement (k = 0.868) between molecular diagnostic tests. Latent class analysis showed that all serological tests were 100 % sensitive, in contrast to the molecular tests with 47 % sensitivity. All tests, though, were highly specific (≥ 97 %). Given the persistence of circulating antibodies after cure, detectable by serological tests, it is recommend combining a serological and a molecular diagnostic test for accurate diagnosis of infection with T. evansi in domestic animals.

Oral flora of stray dogs and cats in Algeria: Pasteurella and other zoonotic bacteria.

BACKGROUND AND AIM: Knowledge of potentially pathogenic bacteria presents in the oral cavity of dogs and cats may be helpful in determining appropriate treatment for infected bite wounds. About 120.000 people are exposed to dog and cat bites every year in Algeria, but little is known about the dog and cat oral flora causing bite wound complications. The purpose of this study was to identify potential zoonotic bacteria from oral cavity of dogs and cats and to determine their susceptibility to antibiotics to contribute to the treatment of bite wound infection. MATERIALS AND METHODS: Oral swabs from 100 stray dogs and 100 stray cats were collected and cultured in several media: Chocolate agar, MacConkey agar, and Mannitol Salt Agar. Bacterial isolates were identified using several commercial kits of the analytical profile index and tested for antibiotic susceptibility by disk diffusion method. RESULTS: Overall, 185/200 (92.5%) dogs and cats carried zoonotic bacteria in their mouths, of which 55.13% (102/185) had at least two bacterial pathogens. 374 pathogenic strains belonging to 15 genera were isolated: Eleven were Gram-negative (Proteus, Pasteurella, Escherichia, Moraxella, Klebsiella, Acinetobacter, Enterobacter, Pseudomonas, Aeromonas, and Neisseria Haemophilus) and four were Gram-positive (Staphylococcus, Streptococcus, and Corynebacterium, Bacillus). Fifty-one strains of Pasteurella were isolated from 44 carriers of Pasteurella (21 Pasteurella multocida, 21 Pasteurella pneumotropica, and 9 Pasteurella spp.). Pasteurella strains were tested for antibiotic resistance. Resistance to at least one drug was observed in 8 (15.68%) of Pasteurella isolates and two strains (3.92%) were found to be multidrug-resistant (to two or more drugs). Erythromycin, penicillin, and ampicillin were the antimicrobials to which the isolates showed greater resistance (7.84%, 5.88%, and 3.92%, respectively). CONCLUSION: To the best of our knowledge, this study is the first in Algeria to detect potential human pathogenic bacteria in the oral cavity of dogs and cats. It reveals that these animals have multiple zoonotic bacteria in their mouths including Pasteurella species, which may be multidrug-resistant.

Epidemiological study of sausage in Algeria: Prevalence, quality assessment, and antibiotic resistance of Staphylococcus aureus isolates and the risk factors associated with consumer habits affecting foodborne poisoning.

AIM: The first aim was to assess the quality and determine the prevalence and antibiotic susceptibility of Staphylococcus aureus contamination of raw sausage sold in ten municipalities in the Northeast of Algeria. Second, a consumer sausage purchasing survey was designed to investigate potential risk factors that have a significant association with the occurrence of foodborne poisoning among sausage consumers' behavior and its relationship with independent variables. MATERIALS AND METHODS: A total of 230 butcheries from ten departments (Daira) of Algiers with more than 40 municipalities were included randomly in these studies to collect raw sausage samples and to distribute 700 structured questionnaires to meat consumers. Our two studies were conducted at the same time, between June 2016 and April 2018. Sausage samples were taken once per butchery to estimate the prevalence of S. aureus contamination and therefore deduct the quality assessment of raw sausage (Merguez) sold in Algiers, Algeria. All isolated strains were tested for their antimicrobial resistance. Furthermore, questionnaires were distributed and used to collect information on various aspects of sausage consumption and foodborne disease. The data collected were analyzed with different statistical approaches, such as the Chi-square test and the odds ratio (OR) univariable logistic model. All the risk factors were analyzed by studying their association with the occurrence of consumers who claimed to have food poisoning after consuming sausage. RESULTS: The overall prevalence of S. aureus contamination from sausages was 25.22% (n=58/230). Over 83.33% of strains showed resistance to at least one of the antibiotics tested. The most important was for tetracycline (58%) followed by fosfomycin (33%), penicillin G (25%), and oxacillin (36%). Moreover, the multiple antibiotic resistance (MAR) index include 20 profiles with MAR >0.2. Out of the 440 meat consumers, 22.16% revealed having food poisoning after sausage consumption. The risk factors recorded were: Consumption outside of home (24.30%, OR=1.769, p=0.040), during the summer season (24.30%, OR=1.159) and during lunch (26.50%, OR=1.421). CONCLUSION: Our study highlights a high prevalence of S. aureus contamination in Merguez, especially in some departments of Algiers, and the high multidrug resistance of S. aureus isolates against tetracycline and oxacillin; thus, S. aureus contamination in sausage is considered a potential risk to public health. Therefore, to reduce and prevent the spread of resistant strains, robust management and monitoring of antibiotic use should be established. Therefore, it is necessary to improve the sanitation conditions and education regarding personal hygiene and change certain consumption habits of Algerian consumers to ensure food safety. Finally, it can be concluded that the application of the HACCP system is essential either in butcheries producing sausage and/or slaughterhouses. From this perspective, studies might be performed to characterize Staphylococcus spp and S. aureus to investigate their virulence factors.

Serological, clinical, and risk factors of the Newcastle disease on broilers flocks in Algeria.

AIM: The work aimed at studying the serological and clinical factors, as well as the risk factors of the Newcastle disease (ND) on broilers herds in Algeria. MATERIALS AND METHODS: A sample of 1248 birds was randomly selected from 52 broiler flocks. We took blood samples from each bird at the level of the wing vein area where an indirect enzyme-linked immunosorbent assay technique was carried out through the use of an IDvet kit. RESULTS: The flocks showed 82.69% of seroprevalence. Clinically speaking, the most common symptoms were sneezing, rale, greenish diarrhea, torticollis, and motor discords. Most commonly observed postmortem lesions were the proventriculitis, tracheitis, and enteritis. Especially, the caeca are hemorrhagic. The scores show the effect of risk factors. There was a significant effect on the mortality, the hygiene and vaccination groups on antibody titers in time 2. The antibody titers were elevated in the herd that recorded a high mortality (more than 10%) compared with those which recorded a low mortality (<10%) (p=0.002). Therefore, the antibody titers were elevated in herds with bad hygiene, compared with the ones with good hygiene (p=0.04). At last, when broiler chicken were not boosted by ND vaccine, flocks appeared to be more seropositive (p=0.02). CONCLUSION: The serological survey conducted in this study provided an important scope for ND as a dominant viral disease in broilers. Many factors are responsible for the onset of these diseases; correct biosecurity measures are needed to reduce the impact of this pathology in poultry farms.

Presence of Rickettsia aeschlimannii, 'Candidatus Rickettsia barbariae' and Coxiella burnetii in ticks from livestock in Northwestern Algeria.

Livestock and their ectoparasites are involved in the epidemiology of several zoonotic diseases. Studies regarding the molecular detection of infectious agents in ticks from Northwestern Algeria are scarce. Thus, the presence of spotted fever group Rickettsia spp., Anaplasmataceae microorganisms and Coxiella burnetii was investigated in ticks collected from ruminants in Sidi Bel Abbes and Saida provinces. Rickettsia aeschlimannii was detected in one Hyalomma excavatum pool and one H. marginatum pool. Moreover, 'Candidatus Rickettsia barbariae' was found in one H. excavatum and six Rhipicephalus bursa pools. Lastly, Coxiella burnetii was amplified in two H. excavatum and two R. bursa pools. No Anaplasmataceae bacterium was detected. This study demonstrates the presence of the tick-associated microorganism 'Candidatus R. barbariae' in the North of Africa, and corroborates the presence of the zoonotic pathogens R. aeschlimannii and C. burnetii in Algeria.

Urinary shedding of pathogenic Leptospira in stray dogs and cats, Algiers: A prospective study.

BACKGROUND: Leptospirosis is an important worldwide zoonosis. This disease is caused by pathogenic species of the genus Leptospira which are maintained in the environment via chronic renal infection of carrier animals which can be asymptomatic excretors of the organisms in their urines and become a source of infection for humans and other hosts. The prevalence of animal leptospirosis in Algiers, Algeria, is unknown. METHODOLOGY/PRINCIPAL FINDINGS: Real-time PCR and standard PCR and sequencing were used to detect pathogenic Leptospira organisms in the urines of stray dogs and cats in Algiers. In the presence of appropriate controls, none of the 107 cat urine samples were positive while 5/104 (4.8%) canine urine samples (asymptomatic mixed-breed dogs, three females and two males) were positive in two real-time PCR assays targeting the rrs and hsp genes. The positivity of these samples was confirmed by partial PCR-sequencing of the rpoB gene which yielded 100% sequence similarity with Leptospira interrogans reference sequence. In this study, L. interrogans prevalence was significantly higher in dogs aged < one year (16.46% - 29.41%) than in adults (0%) (P value = 0.0001) and then in the overall dog population (2.68% - 4.8%) (P = 0.0007). CONCLUSIONS/SIGNIFICANCE: These results suggest that dogs are maintenance hosts for zoonotic leptospirosis in Algiers, Algeria. To face this situation, effective canine vaccination strategies and raising public health awareness are mandatory. Further investigations incorporating a larger sample in more localities will be undertaken to document the epidemiology of urban animal leptospirosis in Algeria at large.

Antimony susceptibility of Leishmania isolates collected over a 30-year period in Algeria.

BACKGROUND: In Algeria, the treatment of visceral and cutaneous leishmanioses (VL and CL) has been and continues to be based on antimony-containing drugs. It is suspected that high drug selective pressure might favor the emergence of chemoresistant parasites. Although treatment failure is frequently reported during antimonial therapy of both CL and VL, antimonial resistance has never been thoroughly investigated in Algeria. Determining the level of antimonial susceptibility, amongst Leishmania transmitted in Algeria, is of great importance for the development of public health policies. METHODOLOGY/PRINCIPAL FINDINGS: Within the framework of the knowledge about the epidemiology of VL and CL amassed during the last 30 years, we sampled Leishmania isolates to determine their susceptibility to antimony. We analyzed a total of 106 isolates including 88 isolates collected between 1976 and 2013 in Algeria from humans, dogs, rodents, and phlebotomines and 18 collected from dogs in France. All the Algerian isolates were collected in 14 localities where leishmaniasis is endemic. The 50% inhibitory concentrations (IC50) of potassium antimony tartrate (the trivalent form of antimony, Sb(III)) and sodium stibogluconate (the pentavalent form of antimony, Sb(V)) were determined in promastigotes and intramacrophage amastigotes, respectively. The epidemiological cutoff (ECOFF) that allowed us to differentiate between Leishmania species causing cutaneous or visceral leishmaniases that were susceptible (S+) or insusceptible (S-) to the trivalent form of antimony was determined. The computed IC50 cutoff values were 23.83 µg/mL and 15.91 µg/mL for VL and CL, respectively. We report a trend of increasing antimony susceptibility in VL isolates during the 30-year period. In contrast, an increase in the frequency of S- phenotypes in isolates causing CL was observed during the same period. In our study, the emergence of S- phenotypes correlates with the inclusion of L. killicki (syn: L. tropica) isolates that cause cutaneous leishmaniasis and that have emerged in Algeria during the last decade. CONCLUSION/SIGNIFICANCE: Our results provide insight into the spatiotemporal dynamics of Leishmania antimony susceptibility in Algeria. We highlight the need for the future implementation of an effective methodology to determine the antimony susceptibility status of Leishmania isolates to detect the emergence of and prevent the dissemination of drug-resistant strains.

Seroprevalence and risk factors for Coxiella burnetii, the causative agent of Q fever in the dromedary camel (Camelus dromedarius) population in Algeria.

Query (Q) fever is a globally distributed zoonotic disease caused by Coxiella burnetii, a bacterial agent for which ruminants are the most prevalent natural reservoir. Data regarding Q fever infection in camels in Algeria are limited. Therefore, a survey to detect seroprevalence of C. burnetii antibodies was conducted among healthy camel populations in a vast area in southeastern Algeria to determine distribution of the Q fever causative organism and to identify risk factors associated with infection. Between January and March 2016, blood samples were collected from 184 camels and serum samples were subsequently analysed using a commercial Enzyme-Linked Immunosorbent Assay (ELISA) kit. At the time of blood collection, a questionnaire investigating 13 potential predisposing factors associated with C. burnetii seropositivity was completed for every dromedary camel and herd. Results were analysed by a chi-square (χ2) test and multivariate logistic regression. The seroprevalence of C. burnetii at the animal level was 71.2% (95% CI: 65.2-78.3) and 85.3% (95% CI: 72.8-97.8) at the herd level. At the animal level, differences in seroprevalence were observed because of herd size, animal age, animal sex, presence of ticks and contact with other herds. A multivariable logistic regression model identified three main risk factors associated with individual seropositivity: (1) age class > 11 years (OR = 8.81, 95% CI: 2.55-30.41), (2) herd size > 50 head (OR = 4.46, 95% CI: 1.01-19.59) and (3) infestation with ticks (OR 2.2; 95% CI: 1.1-4.5). This study of seroprevalence of C. burnetii infection in camels in Algeria revealed a high seroprevalence of Q fever in camel populations in southeastern Algeria and provided strong evidence that Q fever represents an economic, public health and veterinary concern. Appropriate measures should be taken to prevent the spread of C. burnetii and to reduce the risk of Q fever in farm animals and humans in this agro-ecologically and strategically important region of North Africa.

First genetic characterization of Toxoplasma gondii in stray cats from Algeria.

Toxoplasmosis is a parasitic disease with worldwide distribution and a major public health problem. In Algeria, no data are currently available about genotypes of Toxoplasma gondii isolated from animals or humans. The present study assesses for the first time the seroprevalence of toxoplasmosis in stray cats, and provides molecular characterization of T. gondii strains circulating in this feline population in Algiers, the capital city of Algeria. Sera from 96 stray cats were tested for the presence of antibodies against T. gondii using the modified agglutination test. The seroprevalence was 50% (48/96) using 1:6 as the positivity cut-off. Different organs samples from stray cats, including heart samples, were tested for the presence of Toxoplasma DNA using real-time PCR. T. Gondii DNA was detected in 90.6% (87/96) of hearts. Of these parasitic DNAs, 22 were submitted to genotyping through the analysis of 15 microsatellite markers. The identified genotypes (12 of 22) mainly belonged to the type II lineage.

Peste des petits ruminants (PPR): A neglected tropical disease in Maghreb region of North Africa and its threat to Europe.

BACKGROUND: Peste des petits ruminants (PPR) is a contagious disease listed by the World Organisation for Animal health (OIE) as being a specific hazard. It affects sheep, goats, and wild ungulates, and is prevalent throughout the developing world particularly Asia, the Middle East, and Africa. PPR has been targeted for eradication by 2030 by the Food and Agriculture Organization of the United Nations (FAO) and the OIE, after the successful eradication of the related disease, rinderpest in cattle. PPR was first reported in 1942 in the Ivory Coast in Western Africa and has since extended its range in Asia, the Middle East, and Africa posing an immediate threat of incursion into Europe, South East Asia and South Africa. Although robust vaccines are available, the use of these vaccines in a systematic and rational manner is not widespread, resulting in this devastating disease becoming an important neglected tropical disease in the developing world. METHODOLOGY: We isolated and characterized the PPR virus from an outbreak in Cheraga, northern Algeria, during October 2015 by analyzing the partial N-gene sequence in comparison with other viruses from the Maghreb region. As well as sequencing the full length viral genome and performing real-time RT-PCR on clinical samples. Maximum-likelihood and Bayesian temporal and phylogeographic analyses were performed to assess the persistence and spread of PPRV circulation from Eastern Africa in the Maghreb region of North Africa. CONCLUSIONS: Recent PPR outbreaks in Cheraga, in the northern part of Algiers (October 2015) and North-West Morocco (June, 2015) highlight that PPRV has spread to the northern border of North Africa and may pose a threat of introduction to Europe. Phylogeographic analysis suggests that lineage IV PPRV has spread from Eastern Africa, most likely from the Sudan 2000 outbreak, into Northern Africa resulting in the 2008 Moroccan outbreak. Maximum-likelihood and Bayesian analysis shows that these North African viruses cluster closely together suggesting the existence of continual regional circulation. Considering the same virus is circulating in Algeria, Morocco and Tunisia, implementation of a common Maghreb PPR eradication strategy would be beneficial for the region.

Development of a Murine Infection Model with Leishmania killicki, Responsible for Cutaneous Leishmaniosis in Algeria: Application in Pharmacology.

In Algeria, Leishmania infantum, Leishmania major, and Leishmania killicki (Leishmania tropica) are responsible for cutaneous leishmaniosis. We established a murine model of L. killicki infection to investigate its infective capacity, some immunophysiopathological aspects, and its suitability for pharmacological purposes. Following the injection of L. major or L. killicki metacyclic promastigotes in the ear dermis of BALB/c mice, the course of infection was followed. The infection with L. killicki caused slower lesion formation than with L. major. The presence of L. killicki or L. major DNA and parasites was detected in the ear dermis and in lymph nodes, spleen, and liver. Lesions induced by L. killicki were nonulcerative in their aspect, whereas those caused by L. major were highly ulcerative and necrotic, which matches well with the lesion phenotype reported in humans for L. killicki and L. major, respectively. The treatment of L. killicki lesions by injection of Glucantime® significantly reduced the lesion thickness and parasite burden. Ear dermal injection of BALB/c mice constitutes a model to study lesions physiopathology caused by L. killicki and presents interest for in vivo screening of new compounds against this pathogen, emerging in Algeria.

Molecular evidence of vector-borne pathogens in dogs and cats and their ectoparasites in Algiers, Algeria.

In Algeria, only limited information is currently available on the prevalence of emergent canine and feline vector-borne diseases. The aim of the present work was to detect by qPCR vector-associated bacteria in stray dogs and cats and their ectoparasites from Algiers. 18/117 (15.38%) dogs and 2/107 (1.87%) cats were positive for at least one vector-borne agent. Coxiella burnetii and Bartonella henselae were identified in 1/117 (0.85%) dog individually. Ehrlichia canis DNA was detected in 17/117 (14.52%) dogs. 1/107 (0.93%) cat was positive to C. burnetii and another 1/107 (0.93%) to B. henselae. DNA of Rickettsia massiliae, Rickettsia conorii and E. canis was detected in Rhipicephalus sanguineus. Cat fleas were infected with Rickettsia felis, B. henselae and Bartonella clarridgeiae. B. vinsonii subsp. berkhoffii was identified in Xenopsylla cheopis collected from dogs. The findings of this study indicate that dogs and cats from Algeria are exposed to multiple tick and flea-borne pathogens.

Transmission potential of antimony-resistant leishmania field isolates.

We studied the development of antimony-resistant Leishmania infantum in natural vectors Lutzomyia longipalpis and Phlebotomus perniciosus to ascertain the risk of parasite transmission by sand flies. All three resistant strains produced fully mature late-stage infections in sand flies; moreover, the resistant phenotype was maintained after the passage through the vector. These results highlight the risk of circulation of resistant Leishmania strains and question the use of human drugs for treatment of dogs as Leishmania reservoirs.

Antimony resistance and environment: Elusive links to explore during Leishmania life cycle.

Leishmania drug resistance and particularly antimony resistance still continues to emerge in different part of the world. Because visceral and cutaneous leishmaniasis are transmitted in foci with zoonotic or anthroponotic life-cycles, the link between chemotherapeutic resistance and the selection for drug resistance, through drug consumption, cannot be as obvious for all forms of leishmaniasis. The underlying factors that trigger the selection of antimony resistant parasites are poorly studied in regard to environmental aspects. Recently, a correlation between the emergence of antimony unresponsiveness in India and water arsenic contamination has been raised. The presence of some yet unidentified environmental factors driving the selection of antimony resistant Leishmania populations in a zoonotic context of leishmaniasis is also currently questioned. The identification of key molecules involved in the selection of antimony resistance and their importance in the selective process have to be re-evaluated in light of the environment were all the hosts of Leishmania (mammalian and arthropod) evolved. These new insights will help to (i) address the risk of therapeutic failure associated with the emergence of drug-resistance and (ii) propose new therapeutic protocols to aim at reducing the risk of resistance in endemic areas.

Leishmania antimony resistance: what we know what we can learn from the field.

Leishmania is the causative agent of various forms of leishmaniasis, a significant cause of morbidity and mortality. The clinical manifestations of the disease range from self-healing cutaneous and mucocutaneous skin ulcers to a fatal visceral form named visceral leishmaniasis or kala-azar. In the absence of any effective vaccine, the only means to treat and control leishmaniasis is affordable medication. The treatment choice is essentially directed by economic considerations; therefore, for a large majority of countries, chemotherapy relies only on the use of cheaper antimonial compounds. The emergence of antimonial therapy failure in India linked to proven parasite resistance has stressed questions about selective factors as well as transmission risk of drug resistance. Unfortunately, in most parts of the world, the frequency of parasite antimony resistance linked to treatment failure is unknown because of a lack of information on Leishmania antimony susceptibility. This information is crucial for addressing the risk of selection and transmission of drug-resistant parasites, particularly in areas where antimony is the only chemotherapeutic alternative. However, the poor knowledge about factors that favor selection of resistant parasites, the multiplicity of the agents that can play a role in the in vivo antileishmanial activity of antimony, and the lack of a standard protocol to diagnose and survey parasite resistance all contribute to insufficient monitoring of antimony resistance. In this review, we discuss on the factors potentially involved in the selection of antimony resistance in the field and discuss on the methods available for its diagnosis.