Neutralizing based seroprevalence study of Toscana virus in livestock from Algeria.

Toscana virus is a sandfly-borne human pathogen belonging to Phlebovirus genus into Phenuiviridae family. It is emerging in north Africa posing a complex threat to public health. TOSV is heavily affecting sandfly-exposed people in northern Algeria. A larger distribution has recently been stated in Algeria by using dog sera. Dog exposure to TOSV was repeatedly identified in north Algeria, with 4.56% lately detected to possess respective neutralizing antibodies. However, evidence for TOSV has only been observed in dogs among various species of domestic animals. Therefore, we attempted to assess sera from 221 livestock comprising cattle, sheep, goats, rabbits and horses, to identify the presence of TOSV neutralizing antibodies. The study was conducted during 2017, in 11 areas from the governorates of Blida, Medea, Algiers, Tipaza, Ain Defla, Tissemsilt in the north center, and Setif, Mila, Tizi Ouzou, Jijel in the northeast of Algeria. Positive results were obtained in 14.6% (12/82) cattle, 17.18% (11/64) sheep, 15% (3/20) horses and 3.33% (1/30) goats, whereas rabbits remained negative. Positive samples originated mainly from the north centre, with new areas being first-ever detected. The seroprevalence was noticed to be very strongly related to sample origin (p < 0.01). Females (OR=4.09) were observed to be more likely infected. Our findings represent a further proof of TOSV circulation in Algeria. Moreover, they revealed a potential role of livestock (p = 0.00731) in its natural cycle. This fact emphasize how important is to elucidate the exact contribution of livestock to the epidemiology of sandfly-borne phleboviruses, and their impact on public health.

Serological evidence of Rift Valley fever viral infection among camels imported into Southern Algeria.

Rift Valley fever (RVF) is a mosquito-borne viral zoonosis caused by the Rift Valley fever virus (RVFV). The present work aims to investigate the epidemiological status and identify the risk factors associated with RVFV infection in dromedary camels (Camelus dromedarius) from southern Algeria. A total of 269 sera of apparently healthy camels was collected and tested using a competitive Enzyme-Linked Immunosorbent Assay (ELISA). Overall, 72 camels (26.7 %, 95 % CI: 21.4-32) were seropositive to RVFV. IgG antibodies were found to be most prevalent in camels from south-western areas, particularly in Tindouf wilaya (52.38 %, p < 0.0001), and in camels introduced from bordering Sahelian countries (35.8 %) (OR = 8.75, 95 %CI: 2.14-35.81). No anti-RVFV antibodies were detected in sera collected from local camels (0 %). Adult (5-10 years) and aged (>10 years) camels have a significantly higher risk of being infected by RVFV (OR = 2.15; 95 %CI = 1.21-3.81, OR = 2.05; 95 %CI = 1.03-4.11, respectively). This report indicated that dromedaries imported to the south-western areas are exposed to RVFV and may contribute to its spread in Algerian territories.

Assessing the Effectiveness of Qista Baited Traps in Capturing Mosquito Vectors of Diseases in the Camargue Region (France) and Investigating Their Diversity.

Nuisance, allergy, and vector role: mosquitoes are responsible for numerous inconveniences. Several strategies have been employed to fight against this confirmed vector. To record the diversity of mosquito vectors in Camargue (France) and assess the effectiveness of the Qista trap, six BAMs were deployed as a belt barrier to protect the Espeyran Castle (Saint-Gilles, Camargue). Prior to evaluating the reduction in the nuisance rate, recovery nets from the traps and human landing catches (HLC) were utilized twice a week in the treated and control areas. Overall, 85,600 mosquitoes were captured, belonging to eleven species, namely Aedes albopictus, Aedes caspius, Aedes detritus, Aedes dorsalis, Aedes rossicus, Aedes vexans, Anopheles maculipennis, Culex pipiens, Culex modestus, Culiseta annulata and Culiseta longiareolata. The six BAM devices trapped 84,461 mosquitoes. The average capture rate per BAM is 76.92 mosquitoes per day. The rate of nuisance has decreased from 4.33 ± 2.88 before the deployment to 1.59 ± 2.77 after BAM implantation. The Qista BAM trap seems to be an excellent tool for reducing the nuisance rate and may help researchers to optimize trapping methods by obtaining more significant sample sizes. It may also allow the updating of the host-seeking mosquito species' reported biodiversity in the south of France.

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.

MALDI-TOF MS Identification of Dromedary Camel Ticks and Detection of Associated Microorganisms, Southern Algeria.

This study used MALDI-TOF MS and molecular tools to identify tick species infesting camels from Tamanrasset in southern Algeria and to investigate their associated microorganisms. Ninety-one adult ticks were collected from nine camels and were morphologically identified as Hyalomma spp., Hyalomma dromedarii, Hyalomma excavatum, Hyalomma impeltatum and Hyalomma anatolicum. Next, the legs of all ticks were subjected to MALDI-TOF MS, and 88/91 specimens provided good-quality MS spectra. Our homemade MALDI-TOF MS arthropod spectra database was then updated with the new MS spectra of 14 specimens of molecularly confirmed species in this study. The spectra of the remaining tick specimens not included in the MS database were queried against the upgraded database. All 74 specimens were correctly identified by MALDI-TOF MS, with logarithmic score values ranging from 1.701 to 2.507, with median and mean values of 2.199 and 2.172 ± 0.169, respectively. One H. impeltatum and one H. dromedarii (2/91; 2.20%) tested positive by qPCR for Coxiella burnetii, the agent of Q fever. We also report the first detection of an Anaplasma sp. close to A. platys in H. dromedarii in Algeria and a potentially new Ehrlichia sp. in H. impeltatum.

First Isolation of Punique Virus from Sand Flies Collected in Northern Algeria.

In the last decade, several phleboviruses transmitted by sand flies were detected in the Mediterranean countries, with the health impact of some of them being unknown. From September to October 2020, a total of 3351 sand flies were captured in Kherrata (Bejaia, northern Algeria) and identified by sex, grouped in 62 pools, which were tested for the presence of phlebovirus RNA using endpoint RT-PCR. Two pools (male and female, respectively) were positive. The genome sequencing and phylogenetic analysis showed that the two phleboviruses detected were closely related to the Punique virus (PUNV) isolated in Tunisia and detected in Algeria. Both PUNV strains were isolated on VERO cells from positive pools. Morphological identification of 300 sand flies randomly selected, showed a clear dominance of Phlebotomus perniciosus (98.67%). The dominance of this species in the study area was confirmed by PCR targeting the mitochondrial DNA. Our result represents the first isolation of PUNV and the second report in Algeria from two distinct regions which confirm its large circulation in the country and more broadly in North Africa. Further studies are needed to measure the impact on public health through seroprevalence studies in humans as well as animals and to investigate its potential involvement in neurological viral diseases.

Presence of the sandfly-borne phlebovirus (Toscana virus) in different bio-geographical regions of Algeria demonstrated by a microneutralisation-based seroprevalence study in owned dogs.

Toscana virus (TOSV) is major meningitis and meningoencephalitis agent in the Mediterranean basin. Dogs are frequently exposed to TOSV; thereby they can contribute to estimating its circulation. In Algeria, little is known about its circulation, and available data are restricted to the Kabylian region. To investigate the current situation in Algeria, a total of 205 dog sera collected from 13 different wilayas over the country were analyzed by using in-house Enzyme Linked Immunosorbent Assay (ELISA) and microneutralization test (MNT). An overall seroprevalence rate of 20% (14.5-25.5%) was observed by ELISA. Whereas, a seroprevalence rate of 4.56% (1.65-7.43%) was recorded by microneutralization test elucidating the exact occurrence of TOSV exposure in dogs, in Algeria. Positive dogs were detected from the areas of Algiers, Bejaia, Blida, Bouira, Medea, Setif, and Tlemcen in the north; Laghouat in the high lands and Tamanrasset in great Sahara. Only one serum, originating from Bejaia in the north east, was positive for both testing methods, while 8/9 positive sera in MNT remained negative in ELISA. MNT negative/ELISA positive result of 40/41 might suggest evidence for dog transmission, and circulation of phleboviruses other than TOSV. Noticeably, TOSV and antigenically related viruses are largely prevalent. Thus, they are not only confined to Kabylia region, but are widespread in Algeria, despite its climate diversity.

Identification of blood source preferences and Leishmania infection in sand flies (Diptera: Psychodidae) in north-eastern Algeria.

Leishmaniases are among the most neglected vector-borne diseases, infecting humans as well various animal hosts with clinical outcomes varying from cutaneous disorders to visceral and life-threatening disease. In Algeria, canine leishmaniasis (CanL) caused by Leishmania infantum is endemic mainly throughout the northern regions of the country with the Mediterranean climate that favours the occurrence of Larroussius sand flies, the vectors of the parasite. This study conducted in Bougaa and Kherrata, two regions located in north-eastern Algeria and endemic for CanL, focuses on: i) composition of sand fly fauna, ii) screening of Leishmania parasites and iii) the blood sources of engorged females. Entomological surveys were conducted between June and September 2019 using CDC light-traps in rural areas of both regions. Sand fly specimens were morphologically identified, females were screened for Leishmania DNA using kDNA and ITS1 primers, blood meals in engorged females were identified by peptide mass mapping (PMM)-based MALDI-TOF mass spectrometry and confirmed by DNA sequencing analysis. Overall, 1940 specimens (844 males, 1096 females) were collected, all belonging to the subgenus Larroussius: Phlebotomus perniciosus, (94.64%), Ph. perfiliewi (4.74%) and Ph. longicuspis (0.62%). No Leishmania DNA was detected in the evaluated pools (n = 106) (1096 females). PMM-based MALDI-TOF MS successfully identified a source of blood in 92% (141/154) of engorged females (135 Ph. perniciosus and 6 Ph. perfiliewi). All blood meals were taken from domestic cattle (Bos taurus) except for one originating from a dog (Canis lupus familiaris) and one from sheep (Ovis aries). Sequencing of host cytochrome B gene confirmed these identifications but showed lower success rate of 58% (29/50), demonstrating the high effectivity of peptide mass mapping (PMM)-based MALDI-TOF mass spectrometry for routine identification of blood meals of varying degree of digestion. Our findings represent first record of cattle and dog blood in sand flies in Algeria and striking feeding preference of local sand fly population at domestic sites of studied regions for cattle which may play an important role in parasite transmission. Further studies are needed to better understand potential contribution of cattle to ecology of sand flies and epidemiology of leishmaniasis in north-eastern Algeria.

Identification of Algerian field-caught mosquito vectors by MALDI-TOF MS.

Vector-borne diseases represent a real threats worldwide, in reason of the lack of vaccine and cure for some diseases. Among arthropod vectors, mosquitoes are described to be the most dangerous animal on earth, resulting in an estimated 725,000 deaths per year due to their borne diseases. Geographical position of Algeria makes this country a high risk area for emerging and re-emerging diseases, such as dengue coming from north (Europe) and malaria from south (Africa). To prevent these threats, rapid and continuous surveillance of mosquito vectors is essential. For this purpose we aimed in this study to create a mosquito vectors locale database using MALDI-TOF mass spectrometry technology for rapid identification of these arthropods. This methodology was validated by testing 211 mosquitoes, including four species (Aedes albopictus, Culex pipiens, Culex quinquefasciatus, and Culiseta longiareolata), in two northern wilayahs of Algeria (Algiers and Bejaia). Species determination by MALDI TOF MS was highly concordant with reference phenotypic and genetic methods. Using this MALDI-TOF MS tool will allow better surveillance of mosquito species able to transmit mosquito borne diseases in Algeria.

Phlebotomine sandflies and associated pathogens in Algeria: update and comprehensive overview.

Being the only established vectors of the protozoan parasites of the genus Leishmania, sand flies have become very important in all countries where leishmaniasis exists. It is caused by a variety of species, each one having specific mammalian reservoir hosts and vectors. Leishmania and sand fly classification has always been a controversial matter, and the increasing number of sand fly species described world complicates the task. Until recently, sand flies distributed in the Old World were known as vectors for few Phleboviruses including two known species (Sandfly fever Naples virus, and alehebad virus), and tentative species such as sand fly fever Sicilian virus and Corfou virus. These infections are emerging in the Mediterranean region and will likely spread in forthcoming decades, posing a complex threat to public health. Here, we reviewed the current literature on phlebotomine sand flies fauna and epidemiology of sand fly­borne infections in Algeria.

Epidemiological situation analysis of cutaneous leishmaniasis in Batna (northeast): An important focus in Algeria.

Cutaneous leishmaniasis (CL) is a parasitic infection which poses a real public health problem in various countries, including Algeria. We aimed in this study to investigate the current epidemiological situation of CL in the wilaya of Batna (northeastern Algeria) during 2010-2020 and to discuss some control measures used to combat the disease in the most endemic foci. Data on CL cases and control measures were obtained from the "Direction de la Santé et de la Population (DSP) of Batna" and the "Etablissement Public de la Santé de Proximité (EPSP) of Barika". Data were analyzed and mapped using Microsoft Office Excel 2007, SPSS.21, and ArcGIS 10.3. A total of 9259 cases were notified, with a peak incidence of 1924 cases in 2010. The annual evolution of the disease showed decreasing and increasing trends during the study period. Cases were detected year-round, with a peak notification in November (1957 cases) and January (1663 cases). CL affects both genders and all age groups with predominance for males (58.6%) and infants less than 9 years old (54.4%). Most cases have occurred in western areas of the wilaya especially in the commune of Barika with 2979 cases (32.17%), followed by Abdelkader Azil (13.37%), Bitam (12.32%), Djezzar (8.67%), Ouled Ammar (7.07%), and Amdoukal (4.36%) communes. Spatiotemporal analysis showed that the highest frequencies of CL were exclusively reported in the western regions. There was no evidence of a possible relationship between Deltamethrin house spraying and leishmaniasis incidence. This study revealed that CL is highly endemic in the western areas of Batna, especially in Barika, with children less than 9 years old as the most age group at risk. The development of control measures and the selection of appropriate strategies are necessary to prevent, control, and eradicate the disease.

Bacterial Infections in Humans and Nonhuman Primates from Africa: Expanding the Knowledge.

The close phylogenetic relationship between humans and other primates creates exceptionally high potential for pathogen exchange. The surveillance of pathogens in primates plays an important role in anticipating possible outbreaks. In this study, we conducted a molecular investigation of pathogenic bacteria in feces from African nonhuman primates (NHPs). We also investigated the pathogens shared by the human population and gorillas living in the same territory in the Republic of Congo. In total, 93% of NHPs (n=176) and 95% (n=38) of humans were found to carry at least one bacterium. Non-pallidum Treponema spp. (including T. succinifaciens, T. berlinense, and several potential new species) were recovered from stools of 70% of great apes, 88% of monkeys, and 79% of humans. Non-tuberculosis Mycobacterium spp. were also common in almost all NHP species as well as in humans. In addition, Acinetobacter spp., members of the primate gut microbiota, were mainly prevalent in human and gorilla. Pathogenic Leptospira spp. were highly present in humans (82%) and gorillas (66%) stool samples in Congo, but were absent in the other NHPs, therefore suggesting a possible gorillas-humans exchange. Particular attention will be necessary for enteropathogenic bacteria detected in humans such as Helicobacter pylori, Salmonella spp. (including S. typhi/paratyphi), Staphyloccocus aureus, and Tropheryma whipplei, some of which were also present in gorillas in the same territory (S. aureus and T. whipplei). This study enhances our knowledge of pathogenic bacteria that threaten African NHPs and humans by using a non-invasive sampling technique. Contact between humans and NHPs results in an exchange of pathogens. Ongoing surveillance, prevention, and treatment strategies alone will limit the spread of these infectious agents.

Potential of Artesunate in the treatment of visceral leishmaniasis in dogs naturally infected by Leishmania infantum: Efficacy evidence from a randomized field trial.

Leishmaniasis is among the world's most neglected diseases. Dogs are the main reservoirs/hosts of Leishmania infantum, causative agent of both canine and human visceral leishmaniosis. Canine leishmaniasis (CanL) represents a public health problem as one of the most prevalent zoonotic diseases worldwide. Current therapeutics present drawbacks; thus, there is a need for more effective, safer, and cheaper drugs. The aim of this study was to evaluate and to compare the efficacy of oral administration of artesunate or meglumine antimoniate/allopurinol in dogs with clinical leishmaniasis. Forty-two dogs with naturally occurring clinical leishmaniasis were included in this open-label, simple randomized positive-control clinical field trial with 6 months of follow-up. Dogs received meglumine antimoniate 100 mg/kg/day and allopurinol 30 mg/kg/day for 28 days (control group, n = 26) or artesunate 25 mg/kg/day for 6 days (test group, n = 16). The animals were evaluated for their clinical evolution, parasite load (by qPCR) and humoral response at different time points: 0, 30, 90, and 180 days after treatment. Data analyses showed a significant improvement in both groups in clinical scores, parasitemia and antibody titers after treatment. Compared to the control group, the artesunate group showed significantly lower clinical score (P = 0.0001), lower parasitemia (P = 0.0001) and antibody titers after 6 months of follow-up. Compared to baseline values, a rapid, significant reduction (P < 0.012) in antibody levels, 2.28- versus 3.04-fold for the control versus artesunate groups, respectively, was observed 30 days after treatment. Antibody levels continued to decrease further in the artesunate group, where 58% of cases became seronegative at the 6-month follow-up. All qPCR-positive dogs were negative after treatment with artesunate, while 14.3% remained positive with the appearance of two new cases in the control group. Artesunate was well tolerated, and no side effects were recorded. Treatment failures were similar in both groups with 27.27% (6/22), including 18.18% (4/22) mortality in the control group, versus 26.66% (4/15), including 13.33% (2/15) mortality in the artesunate group. This is the first report showing the potential of artesunate in the treatment of dogs with clinical leishmaniasis. Artesunate showed higher efficacy than the current first-line treatment for CanL without any adverse effects. It could be a good alternative chemotherapy for CanL, and may be considered for further studies in human leishmaniases. Further clinical trials are needed to confirm these findings, to determine if there are relapses after treatment and if dogs remain infective to sandflies, to define the ideal therapeutic dosage and duration of treatment with artesunate.

Canine vector-borne protozoa: Molecular and serological investigation for Leishmania spp., Trypanosoma spp., Babesia spp., and Hepatozoon spp. in dogs from Northern Algeria.

Dogs are competent reservoirs/hosts of several protozoan pathogens transmitted by blood-feeding arthropods. Throughout their long history of domestication, they have served as a link for the exchange of parasites among livestock, wildlife, and humans and therefore remain an important source of emerging and re-emerging diseases. In Algeria, while canine leishmaniosis (CanL) is well known to be endemic, no data are available on other vector-borne protozoans. Here, we investigate the occurrence and diversity of trypanosomes, piroplasms and Hepatozoon spp. and update the epidemiological status of CanL in dogs from Kabylia, northern Algeria. A total of 227 dogs from three regions of Kabylia were enrolled, including 77 dogs with clinical signs. Dogs were clinically examined and were tested for L. infantum antibodies using a Rapid Immuno-Migration (RIM™) and a quantitative indirect Immunofluorescence Antibody Test (IFAT). PCR screening and sequencing were performed for vector-borne protozoa. Sixty two percent (141/227) of dogs presented at least one infection, whereas 26% (59/227) were co-infected. L. infantum antibodies were detected in 35.7% (81/227) of dogs including 88.7% (68/77) of sick dogs. Molecular investigation revealed prevalence of: 6.6% (15/227), 13.2% (30/227), 41% (93/227) for Trypanosoma spp., B. vogeli and H. canis, respectively. T. evansi (3.1%) and potential new subspecies of T. congolense had been identified. Dog''s clinical status correlates positively with L. infantum antibody titers and the presence of co-infections. Susceptibility to CanL varied according to the dog's aptitude and guard dogs were more infected (51%) (P-value = .001). B. vogeli infection was more frequent in juveniles than adults (32% vs 9%, P-value < .001) and in females than males (21% vs 10%, P-value = .02). To the authors' knowledge, this is the first report on vector-borne protozoa infected dogs in Algeria. Current results are important not only for animal health, but also to avoid serious public health and livestock problems.

First serological evidence of West Nile virus infection in wild birds in Northern Algeria.

While the epidemiology of Flaviviruses has been extensively studied in most of the Mediterranean basin, little is known about the current situation in Algeria. In order to detect the circulation of West Nile (WNV) and Usutu viruses (USUV) in Kabylia, 165 sera were collected from two wild birds species, namely the long distance migrant Turdus philomelos (song thrush) (n = 92) and the resident Passer domesticus (house sparrow) (n = 73). A total of 154 sera were first analyzed by commercial competition ELISA. WNV and USUV micro-neutralization tests were performed on all c-ELISA positive sera and all samples with poor volume. Overall, 7.8 % (CI95 %: 3.5-11.9) were positive by c-ELISA. Positive results were detected in 12.5 % (CI95 %:5.6-19.4) of song thrushes and 1.5 % (CI95 %: 0.0-4.5) for sparrow. Micro-neutralization tests revealed an overall seroprevalence of 6.7 % for WNV (CI95 %: 2.9-10.3), Neutralizing antibodies were found in 8.7 % (CI95 %: 3.0-14.4) for song thrushes and in 4.1 % (CI95 %: 0.0-8.7) of sparrows. The current study demonstrates significant seroprevalence of WNV antibodies in wild birds in Algeria.

Molecular detection of Leishmania infantum DNA and host blood meal identification in Phlebotomus in a hypoendemic focus of human leishmaniasis in northern Algeria.

BACKGROUND: Leishmania parasites are transmitted by female phlebotomine sand flies that maintain the enzootic cycle by circulating between sylvatic and domestic mammals. Humans are part of this cycle as accidental hosts due to the vector's search for a source of blood. In Algeria, Human Leishmaniases (HL) are endemic and represent a serious public health problem because of their high annual incidence and their spread across the country. The aim of this study is to identify sand fly species fauna (vectors of Leishmania), determine their infection rate and identify their feeding preferences using molecular tools in a hypoendemic focus of HL located in the province of Tipaza, northern Algeria. METHODOLOGY/PRINCIPAL FINDINGS: An entomological survey using CDC light traps was conducted between July and October of 2015 in four HL affected peri-urban locations in the province of Tipaza, northern Algeria. Sand flies were identified using the morphological criteria of the genitalia for the males and spermathecae for the females. Leishmania DNA was detected in pooled female sand flies (N = 81 pools with 8-10 specimens per pool) using quantitative real-time polymerase chain reaction (qPCR) targeting two different genes: kDNA-PCR and 18S rRNA. To identify their blood meal sources, blood-fed female sand flies were analyzed by PCR-sequencing targeting the vertebrate cytochrome c oxidase I (COI) gene. A total of 4,045 sand flies were caught, of which 3,727 specimens were morphologically identified. Seven species were recorded: P. (L.) perniciosus (50.28%), P. (L.) perfiliewi (26.13%), P. (L.) longicuspis (21.92%), Sergentomyia (S.) minuta (0.85%), P. (P.) papatasi (0.42%), P. (L.) langeroni (0.32%) and P. (L.) ariasi (0.05%). Afterwards, 740 female specimens were randomly selected and divided into 81 pools and were then screened to investigate the presence of Leishmania spp. L. infantum DNA was detected in three pools, corresponding to three sand fly specimens (one each). The infection rate was 0.33% (2/600) for P. (L.) perniciosus and 2.56% (1/39) for P. (L.) perfiliewi. Analysis of the blood feeding sources (N = 88 specimens) revealed that sand flies belonging to Larroussius subgenera, mainly (71.5%) feed on small ruminants. Human blood is the second feeding source (17%), eight specimens (9%) were found to feed on equines and no domestic reservoir (dog) blood was found. CONCLUSIONS/SIGNIFICANCE: The presence of human leishmaniasis cases, the high abundance of Phlebotomus (Larroussius) species which are proven or suspected vectors of L. infantum, and the detection of L. infantum DNA from its natural vectors (P. (L.) perniciosus, P. (L.) perfiliewi), in addition to the blood-feeding of positive females for L. infantum on humans blood, prove that the major elements of the epidemiological transmission cycle of L. infantum are present and indicate risk factors for an outbreak of the disease in the province of Tipaza.

Detection of relapsing fever Borrelia spp., Bartonella spp. and Anaplasmataceae bacteria in argasid ticks in Algeria.

BACKGROUND: Argasid ticks (soft ticks) are blood-feeding arthropods that can parasitize rodents, birds, humans, livestock and companion animals. Ticks of the Ornithodoros genus are known to be vectors of relapsing fever borreliosis in humans. In Algeria, little is known about relapsing fever borreliosis and other bacterial pathogens transmitted by argasid ticks. METHODOLOGY/PRINCIPAL FINDINGS: Between May 2013 and October 2015, we investigated the presence of soft ticks in 20 rodent burrows, 10 yellow-legged gull (Larus michahellis) nests and animal shelters in six locations in two different bioclimatic zones in Algeria. Six species of argasid ticks were identified morphologically and through 16S rRNA gene sequencing. The presence and prevalence of Borrelia spp., Bartonella spp., Rickettsia spp. and Anaplasmataceae was assessed by qPCR template assays in each specimen. All qPCR-positive samples were confirmed by standard PCR, followed by sequencing the amplified fragments. Two Borrelia species were identified: Borrelia hispanica in Ornithodoros occidentalis in Mostaganem, and Borrelia cf. turicatae in Carios capensis in Algiers. One new Bartonella genotype and one new Anaplasmataceae genotype were also identified in Argas persicus. CONCLUSIONS: The present study highlights the presence of relapsing fever borreliosis agents, although this disease is rarely diagnosed in Algeria. Other bacteria of unknown pathogenicity detected in argasid ticks which may bite humans deserve further investigation.

Seroprevalence of West Nile virus antibodies in equids in the North-East of Algeria and detection of virus circulation in 2014.

West Nile fever (WNF) is a viral disease of wild birds transmitted by mosquitoes. Humans and equids can also be affected and suffer from meningoencephalitis. In Algeria, since the 1994 epidemic, no data on WNV circulation was available until 2012. In September 2012, a fatal human case of WNV neuro-invasive infection occurred in Jijel province. This study describes the first seroprevalence study of West Nile virus (WNV) antibodies conducted in the equine population in Algeria. During 2014, serum samples were collected from 293 equids (222 donkeys and 71 horses) asymptomatic and unvaccinated for WNV in three localities in Northeastern wetlands of Algeria. Antibodies against WNV were found in 51 samples (seroprevalence 17.4%) of sampled equids, distributed as follows: 19 (seroprevalence 26.8%) horses and 32 (seroprevalence 14.4%) donkeys. Moreover 7 horses coming from Blida, in the center of Algeria, were tested before and after an 8-months stay in North-East Algeria. We observe a seroconversion in 2 horses, showing WNV circulation in 2014 in this specific region of Algeria.

Identification of Algerian Field-Caught Phlebotomine Sand Fly Vectors by MALDI-TOF MS.

BACKGROUND: Phlebotomine sand flies are known to transmit Leishmania parasites, bacteria and viruses that affect humans and animals in many countries worldwide. Precise sand fly identification is essential to prevent phlebotomine-borne diseases. Over the past two decades, progress in matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has emerged as an accurate tool for arthropod identification. The objective of the present study was to investigate the usefulness of MALDI-TOF MS as a tool for identifying field-caught phlebotomine. METHODOLOGY/PRINCIPAL FINDINGS: Sand flies were captured in four sites in north Algeria. A subset was morphologically and genetically identified. Six species were found in these areas and a total of 28 stored frozen specimens were used for the creation of the reference spectrum database. The relevance of this original method for sand fly identification was validated by two successive blind tests including the morphological identification of 80 new specimens which were stored at -80°C, and 292 unknown specimens, including engorged specimens, which were preserved under different conditions. Intra-species reproducibility and inter-species specificity of the protein profiles were obtained, allowing us to distinguish specimens at the gender level. Querying of the sand fly database using the MS spectra from the blind test groups revealed concordant results between morphological and MALDI-TOF MS identification. However, MS identification results were less efficient for specimens which were engorged or stored in alcohol. Identification of 362 phlebotomine sand flies, captured at four Algerian sites, by MALDI-TOF MS, revealed that the subgenus Larroussius was predominant at all the study sites, except for in M'sila where P. (Phlebotomus) papatasi was the only sand fly species detected. CONCLUSION: The present study highlights the application of MALDI-TOF MS for monitoring sand fly fauna captured in the field. The low cost, reliability and rapidity of MALDI-TOF MS analyses opens up new ways in the management of phlebotomine sand fly-borne diseases.

Detection of a novel Rickettsia sp. in soft ticks (Acari: Argasidae) in Algeria.

Argasid ticks are vectors of viral and bacterial agents that can infect humans and animals. In Africa, relapsing fever borreliae are neglected arthropod-borne pathogens that cause mild to deadly septicemia and miscarriage. It would be incredibly beneficial to be able to simultaneous detect and identify other pathogens transmitted by Argasid ticks. From 2012 to 2014, we conducted field surveys in 4 distinct areas of Algeria. We investigated the occurrence of soft ticks in rodent burrows and yellow-legged gull (Larus michahellis) nests in 10 study sites and collected 154 soft ticks. Molecular identification revealed the occurrence of two different soft tick genera and five species, including Carios capensis in yellow-legged gull nests and Ornithodoros occidentalis, Ornithodoros rupestris, Ornithodoros sonrai, Ornithodoros erraticus in rodent burrows. Rickettsial DNA was detected in 41/154, corresponding to a global detection rate of 26.6%. Sequences of the citrate synthase (gltA) gene suggest that this agent is a novel spotted fever group Rickettsia. For the first time in Algeria, we characterize a novel Rickettsia species by molecular means in soft ticks.