Molecular detection and MST genotyping of Coxiella burnetii in ruminants and stray dogs and cats in Northern Algeria.

Aiming at identifying the reservoir and contamination sources of Coxiella burnetii in Northern Algeria, we investigated the molecular presence of the bacterium in 599 samples (blood, placenta, liver, spleen, and uterus) collected from cattle, sheep, dogs and cats. Our qPCR results showed that 15/344 (4.36%) blood samples and six/255 (2.35%) organ specimens were positive for C. burnetii. In cattle, three (4%) blood and liver samples were positive. In sheep, one blood (1.19%) and 3 (8.57%) placenta samples were positive. At the Algiers dog pound, 8 (10%) and 3 (5%) blood samples were qPCR positivein dogs and cats, respectively. In addition, MST genotyping showed that MST 33 was present in cattle and sheep, MST 20 in cattle,andMST 21 in dogs and cats.

TRIzol-based RNA extraction for detection protocol for SARS-CoV-2 of coronavirus disease 2019.

Diagnostic testing is important for managing the 2019 novel coronavirus (SARS-CoV-2). We developed an optimized protocol for SARS-CoV-2 RNA extraction from the surface of the respiratory mucosa with nasopharyngeal swabs and compared the sensitivity of RNA extraction methods. RNA extraction was performed using three different procedures (TRIzol, QIAamp, VMT-TRIzol) from nine positive SARS-CoV-2 samples. SARS-CoV-2 was detected by real-time reverse transcriptase PCR (RT-PCR) using a detection kit for SARS-CoV-2 (Sun Yat-sen University). Compared to RT-PCR results, there were no discernible differences in detection rates when comparing the three different extraction procedures. On the basis of these results, the use of TRIzol as a transport medium and RNA extraction method for SARS-CoV-2 detection may be a helpful alternative for laboratories facing shortages of commercial testing kits.

Seasonal distribution of Rickettsia spp. in ticks in northeast Algeria.

This article discusses research conducted on the sampling of two tick species: Ixodes ricinus and Rhipicephalus bursa. Ticks were collected in northern Algeria (El Tarf) in 2014 and studied for differences in abundance and seasonal distribution of population dynamics, as well as tested by PCR for the presence of Rickettsia spp. By molecular tools, four Rickettsia pathogens agents were detected: R. helvetica, R. monacensis, R. raoultii and R. massiliae.

Corrigendum to 'Coxiella burnetii infection with women's febrile spontaneous abortion reported in Algiers'.

[This corrects the article DOI: 10.1016/j.nmni.2018.08.003.].

Der p 5 allergen from house dust mite: first epitope mapping of rabbit IgG blocking antibodies.

Der p 5 is one of the important house dust mite allergens in Algeria; this allergen is frequently recognized by patients with allergic asthma. However, there is no information on its IgG-binding epitopes. In the present study, rabbits were immunized with recombinant Der p 5 allergen, and serum samples were obtained. Recognition of linear IgG epitopes of Der p 5 was determined using synthesized peptides derived from the allergen sequence. The results showed that serum from immunized rabbits recognized three linear epitopes from Der p 5 (28EDKKHDYQNEFDFLLMERIHEQIK43), (37IHEQIKKGELALFYLQEQ55) and (92LMQRKDLDIFEQYNLEMAKKS112). More interestingly, we observed that the 92L-S112 amino acid sequence is well recognized by both IgE and IgG antibodies. Der p 5 stimulates the synthesis of specific IgG antibodies which recognize common but also novel epitopes compared to IgE antibody binding. Indeed, the potential to induce IgG antibodies can be used to inhibit human IgE binding to allergens which may be part of the mechanism of action of specific immunotherapy.

West Nile virus in Algeria: a comprehensive overview.

West Nile virus (WNV) is a mosquito-borne virus belonging to the genus Flavivirus, related to the Japanese encephalitis antigenic complex of Flaviviridae family. It is transmitted by the bite of infected mosquitoes. The virus is maintained in a mosquito-bird-mosquito transmission cycle. WNV has recently dramatically expanded its geographical range and is now considered the most widespread arbovirus in the world, including the Americas, Europe and countries facing the Mediterranean Basin. In Algeria, West Nile disease (WND) infections with human meningoencephalitis cases have been reported in 1994 in Tinerkouk (southwest Sahara. In autumn 2012, one fatal clinical case of WNV neuroinvasive infection was reported in Jijel (coastal east). During the same year, a retrospective serosurvey performed in Algiers and bordering areas highlighted specific anti-WNV IgG in local population. Between 2013 and 2014 two clinical cases were reported, in Timimoune (south) and Guelma (northeast) respectively. Although no case was reported in equids, serosurveys demonstrated its presence: an animal serosurvey was conducted in Djanet (south) in 1975, and in 2014 a seroprevalence of equids in the northeast part of Algeria highlighted a virus circulation. This review aims to evaluate the global epidemiologic situation of West Nile disease in Algeria, with an updated situation based on human cases, equine reports and entomologic investigations. Our study reinforces the need for building the capacity for surveillance in this region to prevent future emergence of WNV and other arboviruses.

Neglected vector-borne bacterial diseases and arboviruses in the Mediterranean area.

Arthropod vectors can transmit pathogenic microorganisms from one vertebrate to another during their blood meal. Although some vector-borne diseases have been eradicated in the Mediterranean area, such as malaria and dengue, recent endemic microorganisms (Toscana virus, Rickettsia spp.) remain neglected even though they cause many more cases. New diagnostic tools and innovative tools for the identification and characterization of vector species and microorganisms have been developed at IHU Méditerranée Infection, either internally or through collaborative and integrated projects. We have detected Rickettsia slovaca as a human pathogen and have described the disease; we have shown that Rickettsia felis can be transmitted by Anopheles mosquitoes; we have emphasized the increasing importance of bedbug (Cimex lectularius) as a potential vector of Bartonella quintana; and we have described the Toscana virus, a major agent of meningitis and meningoencephalitis which was disseminated in North Africa and Central and Eastern Europe, where it frequently cocirculates with a large number of newly described phleboviruses transmitted by sand flies.

Coxiella burnetii infection with women's febrile spontaneous abortion reported in Algiers.

We investigated Q fever infection in Febrile Spontaneous Abortions in women by using a serologic method (Immuno-Fluorescence Assay, IFA) and a molecular method (real-time quantitative PCR, qPCR) in Obstetric-Gynaecology (OB-GYN) services in two hospitals in Algiers. We included in the case group 380 women who experienced Febrile Spontaneous Abortion; the control group comprised 345 women who gave birth without any other infections or complications. Among the 725 women included, antibodies against Coxiella burnetii were detected by IFA in three (03) cases patients; all control group samples were IFA negative. In other hand, only four (04) placental samples among the case group came back with q PCR positive for IS1111 and IS30a too. A relationship between C. burnetii infection and febrile spontaneous abortion exists in OB-GYN services in Algiers.

Genotyping of Coxiella burnetii detected in placental tissues from aborted dairy cattle in the north of Algeria.

Coxiella burnetii, is an obligate intracellular bacterium which is present throughout the world. In humans, C. burnetii is the causative agent of Q fever. In cattle, the infection is suspected to cause stillbirths, retained fetal membranes, metritis and infertility. The birth products of ruminants shed huge amounts of bacteria, and are considered a major source for human infection. The present study was designed to search for the presence of C. burnetii in placental tissues collected from aborted and normal calving dairy cows in Algeria, using molecular tools. A total of 77 placental tissue fragments were collected from dairy cows. 73 samples were collected from aborted cows and four samples were collected from natural calving cows over a period of two years from January 2013 to March 2015. The presence of C. burnetii in these samples was screened by quantitative real-time polymerase chain reaction (qPCR) targeting two different genes, IS1111 and IS30 A. The positive PCR amplicons were subsequently sequenced for Multispacer Sequence Typing determination (MST) using seven pairs of sequences (Cox2, Cox5, Cox18, Cox37, Cox56, Cox57, and Cox61). Fourteen placental tissues (19.1%) were found to be positive for C. burnetii by qPCR; 9 (12.3%) from the city of Blida and 5 (6.84%) from the city of Medea. Genotyping of the corresponding amplicons displayed 100% identity with C. burnetii MST20 genotype, confirming the circulation of this clone in dairy farms from Algeria.

Molecular evidence of Rickettsia slovaca in spleen of wild boars in northeastern Algeria.

Using molecular assays, Rickettsia slovaca, the agent of a spotted fever group rickettsiosis resulting in scalp eschar and neck lymphadenopathy after tick bite, was assessed in 92 spleens recovered from 117 wild boars hunted in the far northeast of Algeria. Rickettsia slovaca was detected in 5.4% of tested wild boar spleens. The presence of R. slovaca DNA in boar spleens questions the relationship that may exist between this bacterium and Sus scrofa algira, and its role in human infections.

Inventory and update on argasid ticks and associated pathogens in Algeria.

Argasid ticks include vectors of relapsing fevers caused by Borrelia spp. in humans, and they can transmit arboviruses and other bacterial pathogens. Knowledge about soft ticks (Ixodida: Argasidae) in Algeria is incomplete, and distribution data need to be updated. Here we report a series of entomologic investigations that we conducted in five different areas in Algeria between 2012 and 2015. Ticks were identified by entomologic keys and molecular tools (16S rRNA gene). Six distinct species belonging to two genera were identified, including Ornithodoros capensis s.s., Ornithodoros rupestris, Ornithodoros occidentalis, Ornithodoros erraticus, Ornithodoros sonrai and Argas persicus. The present study highlights the distribution of soft ticks, the establishment of an update inventory with nine species and associated pathogens detected in argasid ticks in Algeria.

Acaricidal activity of flavonoids extract of Borago officinalis L. (Boraginaceae) against brown dog tick, Rhipicephalus sanguineus (Latreille, 1806).

Borago officinalis L. (Boraginaceae) is a plant of the Boraginaceae family, used in Algeria for food and medicinal purposes. This study reports the effect of flavonoids extracted from the aerial part of Borago officinalis L. (Boraginaceae) on the larvae and engorged adult females of the brown dog tick Rhipicephalus sanguineus (Latreille, 1806) using adults immersion test (AIT) and larval immersion test (LIT). For this purpose, the larvae and engorged female of Rhipicephalus sanguineus (Latreille, 1806) were exposed to serial dilutions of flavonoids extract (50 mg/ml, 25 mg/ml, 12.5 mg/ml and 6.25 mg/ml) using "dipping method" in vitro. The plant extract was obtained by fractionation using appropriate solvents. The extraction yield is 22% with a flavonoids concentration equal to 129.12 µg equivalent of quercetin/ml of the extract. The chromatographic analysis by high performance thin layer chromatography (HPTLC) reveals the presence of gallic acid, vanillic acid, kaempferol, dihydroxybenzoic and quercetin. The results obtained show that the flavonoids extract of Borago officunalis L. (Boraginaceae) considerably reduces the oviposition and the hatching rate of the eggs of Rhipicephalus sanguineus (Latreille, 1806) and was shown to be toxic against newly hatched larvae of Rhipicephalus sanguineus (Latreille, 1806) (P < 0.05).

First field evidence infection of Culex perexiguus by West Nile virus in Sahara Oasis of Algeria.

BACKGROUND & OBJECTIVES: West Nile virus (WNV) is considered one of the most widely distributed arboviruses in the world which is transmitted by several mosquito species including the Culex genus. Culex pipiens is the major vector of this virus in Europe and USA whereas in African countries, other species such as Cx. perexiguus is considered as an important vector. This paper aimed to study the mosquito species involved in WNV transmission in Aougrout, one of the highly populated Oasis of Timimoun Province in Algeria where human WNV neuroinvasive diseases are prevalent. METHODS: CDC light-traps were installed in animal and human shelters for three nights. Collected mosquitoes were pooled and real-time PCR was performed to detect and identify WNV lineages 1 and 2 in the samples. Results: CDC light-traps collected 270 mosquitoes belonging to three genera. Culex genus was predominant with Cx. perexiguus as main species followed by Aedes and Anopheles genus. A total of 33 pools were tested; one pool containing Cx. perexiguus was found positive for WNV lineage 1. INTERPRETATION & CONCLUSION: This study reports for the first time a WNV natural infection of Culex perexiguus in the study region indicating that species other than Cx. pipiens should be taken into consideration in WNV surveillance, especially in specific environments like Saharan Oasis ecosystem.

Comparative analysis of storage conditions and homogenization methods for tick and flea species for identification by MALDI-TOF MS.

Ticks and fleas are vectors for numerous human and animal pathogens. Controlling them, which is important in combating such diseases, requires accurate identification, to distinguish between vector and non-vector species. Recently, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was applied to the rapid identification of arthropods. The growth of this promising tool, however, requires guidelines to be established. To this end, standardization protocols were applied to species of Rhipicephalus sanguineus (Ixodida: Ixodidae) Latreille and Ctenocephalides felis felis (Siphonaptera: Pulicidae) Bouché, including the automation of sample homogenization using two homogenizer devices, and varied sample preservation modes for a period of 1-6 months. The MS spectra were then compared with those obtained from manual pestle grinding, the standard homogenization method. Both automated methods generated intense, reproducible MS spectra from fresh specimens. Frozen storage methods appeared to represent the best preservation mode, for up to 6 months, while storage in ethanol is also possible, with some caveats for tick specimens. Carnoy's buffer, however, was shown to be less compatible with MS analysis for the purpose of identifying ticks or fleas. These standard protocols for MALDI-TOF MS arthropod identification should be complemented by additional MS spectrum quality controls, to generalize their use in monitoring arthropods of medical interest.

Enzootic plague foci, Algeria.

In Algeria, PCR sequencing of pla, glpD and rpoB genes found Yersinia pestis in 18/237 (8%) rodents of five species, including Apodemus sylvaticus, previously undescribed as pestiferous; and disclosed three new plague foci. Multiple spacer typing confirmed a new Orientalis variant. Rodent survey should be reinforced in this country hosting reemerging plague.

Responses of artificially reared cat fleas Ctenocephalides felis felis (Bouché, 1835) to different mammalian bloods.

The cat flea, Ctenocephalides felis felis (Bouche, 1835) (Siphonaptera: Pulicidae), which is found worldwide and which parasitizes many species of wild and domestic animal, is a vector and/or reservoir of bacteria, protozoa and helminths. To aid in the study of the physiology and behaviour of fleas and of their transmission of pathogens, it would be of value to improve the laboratory rearing of pathogen-free fleas. The conditions under which artificially reared fleas at the University of Bristol (U.K.) and the Rickettsial Diseases Institute (France) are maintained were studied, with different ratios of male to female fleas per chamber (25 : 50, 50 : 100, 100 : 100, 200 : 200). The fleas were fed with bovine, ovine, caprine, porcine or human blood containing the anticoagulants sodium citrate or EDTA. Egg production was highest when fleas were kept in chambers with a ratio of 25 males to 100 females. In addition, the use of EDTA as an anticoagulant rather than sodium citrate resulted in a large increase in the number of eggs produced per female; however, the low percentage of eggs developing through to adult fleas was lower with EDTA. The modifications described in our rearing methods will improve the rearing of cat fleas for research.

Evidence that clade A and clade B head lice live in sympatry and recombine in Algeria.

Pediculus humanus L. (Psocodea: Pediculidae) can be characterized into three deeply divergent lineages (clades) based on mitochondrial DNA. Clade A consists of both head lice and clothing lice and is distributed worldwide. Clade B consists of head lice only and is mainly found in North and Central America, and in western Europe and Australia. Clade C, which consists only of head lice, is found in Ethiopia, Nepal and Senegal. Twenty-six head lice collected from pupils at different elementary schools in two localities in Algiers (Algeria) were analysed using molecular methods for genotyping lice (cytochrome b and the multi-spacer typing (MST) method. For the first time, we found clade B head lice in Africa living in sympatry with clade A head lice. The phylogenetic analysis of the concatenated sequences of these populations of head lice showed that clade A and clade B head lice had recombined, suggesting that interbreeding occurs when lice live in sympatry.

Record of Phlebotomus (Transphlebotomus) mascittii Grassi, 1908 and Phlebotomus (Larroussius) chadlii Rioux, Juminer & Gibily, 1966 female in Algeria.

We report for the first time the presence of Phlebotomus mascittii and the female of Phlebotomus chadlii in Algeria. These two species were collected during an entomological study conducted in endemic visceral leishmaniasis focus from the north part of the country, Kabylia.

First entomological documentation of Aedes (Stegomyia) albopictus (Skuse, 1894) in Algeria.

In August 2010, during an entomological programme targeting sandflies, in the region of Larbaa-Nath-Iraten, Wilaya of Tizi-Ouzou (Algeria), a female Aedes albopictus was trapped alive and partially engorged. To our knowledge, this is the first report of Ae. albopictus in Algeria and more widely in the Maghreb.