High Genetic Diversity and Rickettsia felis in Pediculus humanus Lice Infesting Mbuti (pygmy people), -Democratic Republic of Congo.

Pediculus humanus is an obligate bloodsucking parasite of humans that has two ecotypes, the head louse and the body louse, which share an intimate history of coevolution with their human host. In the present work, we obtained and analysed head and body lice collected from Mbuti pygmies living in the Orientale province of the Democratic Republic of the Congo. Cytochrome b DNA analysis was performed in order to type the six known lice clades (A, D, B, F, C and E). The results revealed the presence of two mitochondrial clades. Clade D was the most frequent (61.7% of 47), followed by clade A (38.3% of 47). Sixteen haplotypes were found in 47 samples, of which thirteen were novel haplotypes, indicating an unusually high genetic diversity that closely mirrors the diversity of their hosts. Moreover, we report for the first time the presence of the DNA of R. felis in three (6.4% of 47) head and body lice belonging to both clades A and D. Additional studies are needed to clarify whether the Pediculus lice can indeed transmit this emerging zoonotic bacterium to their human hosts.

First molecular characterization of Blastocystis subtypes from animals and animal-keepers stool in Algeria.

Blastocystis sp. is one of the most common enteric parasites found in humans and many non-human hosts. It is an anaerobic protozoan that belongs to the group of Stramenopiles. Based on phylogenetic analysis of ribosomal DNA genes, at least 17 subtypes (ST1-ST17) are described. The aim of this study was to identify and characterize Blastocystis sp. in stool samples from various animal groups and animal-keepers. Overall, 29/70 (41.43%) animals and 7/60 (11.66%) humans sampled were positive for Blastocystis sp. using microscopy. The sequencing of the partial 18S small subunit ribosomal DNA gene (SSU rDNA) revealed the presence of five haplotypes corresponding to ST2 and ST3 in humans, and ST2, ST3, ST7, and ST10 in animals. This is the first report of Blastocystis subtypes in animals in Algeria.

New Haplotypes of Trypanosoma evansi Identified in Dromedary Camels from Algeria.

PURPOSE: Surra is a zoonotic disease caused by Trypanosoma evansi (Trypanozoon), a salivary trypanosome native to Africa which affects a wide range of mammals worldwide and causes mortality and significant economic loss. The present study was devoted to the molecular characterization of T. evansi derived from naturally infected dromedary camels in Algeria. METHODS: A total of 148 blood samples were collected from mixed age camels living in one of four geographic regions (Ouargla, El Oued, Biskra and Ghardaia) of Algeria. Samples underwent PCR amplification and sequencing of the internal transcribed spacer 1 (ITS1) complete sequence. RESULTS: DNA of Trypanosoma spp. was found in 19 camels (12.84%). Trypanosoma spp. molecular positivity was not affected by sex (p = 0.50), age (p = 0.08), or geographic location (p = 0.12). Based on multiple sequence alignment of the obtained DNA sequences with representative T. evansi ITS1 sequences available globally, the Algerian sequences were grouped within four different haplotypes including two which were original. CONCLUSION: Results of this study provide preliminary data on which future studies of genetic diversity and molecular epidemiology of T. evansi can be based.

Emergence of mcr-1 plasmid-mediated colistin-resistant Escherichia coli isolates from seawater.

Colistin is currently regarded as one of the 'last-resort' antibiotics used for the treatment of critical infections caused by multidrug-resistant Gram-negative pathogens. Recently, there have been numerous reports of the emergence of a transferable plasmid-mediated colistin resistance gene, mcr-1 in patients, animals, food, and environment. Here, we characterize the support of colistin resistance among environmental isolates collected from seawater of Algiers coast. Our study was carried out on 246 isolates resistant to colistin (MIC > 2 µg/L). The mcr-1 gene was identified in only two isolates; M49 and M78. The two strains were identified as Escherichia coli and were non-susceptible to amoxicillin, ticarcillin, piperacillin, gentamicin, nalidixic acid, tigecycline, tetracycline, trimethoprim-sulfamethoxazole and colistin. For the latter, isolates M49 and M78 showed MIC values of 4 µg/mL and 8 µg/mL, respectively. Only the strain M78 was intermediary resistant to tobramycin. The two E. coli strains belonged to two different sequence types (STs): ST23 for M49 and ST115 for M78. The mcr-1 gene was present on a non-conjugative plasmid in the two strains.

Human Lice in Paleoentomology and Paleomicrobiology.

Lice are a classic example of cospeciation. Human lice confirm this cospeciation with lice specialized in hominids which differ from those of gorillas and chimpanzees. Head lice and body lice seem to belong to closely related species with different ecotypes and a different geographical distribution which may reflect population movements. Paleo-entomology allows us in some cases to trace the migrations of archaic human populations. The analysis of lice found on mummies in Egypt and South America has clarified a certain number of these migrations, also the study of lice and the diseases they transmit has shed a new light on the epidemics of the past.

Host switching of human lice to new world monkeys in South America.

The coevolution between a host and its obligate parasite is exemplified in the sucking lice that infest primates. In the context of close lice-host partnerships and cospeciation, Pediculus mjobergi, the louse of New World primates, has long been puzzling because its morphology resembles that of human lice. To investigate the possibility that P. mjobergi was transmitted to monkeys from the first humans who set foot on the American continent thousands of years ago, we obtained and compared P. mjobergi lice collected from howler monkeys from Argentina to human lice gathered from a remote and isolated village in Amazonia that has escaped globalization. Morphological examinations were first conducted and verified the similarity between the monkey and human lice. The molecular characterization of several nuclear and mitochondrial genetic markers in the two types of lice revealed that one of the P. mjobergi specimens had a unique haplotype that clustered with the haplotypes of Amazonian head lice that are prevalent in tropical regions in the Americas, a natural habitat of New World monkeys. Because this phylogenetic group forms a separate branch within the clade of lice from humans that were of American origin, this finding indicates that human lice have transferred to New World monkeys.

A New Clade of African Body and Head Lice Infected by Bartonella quintana and Yersinia pestis-Democratic Republic of the Congo.

The human body louse is known as a vector for the transmission of three serious diseases-specifically, epidemic typhus, trench fever, and relapsing fever caused by Rickettsia prowazekii, Bartonella quintana, and Borrelia recurrentis, respectively-that have killed millions of people. It is also suspected in the transmission of a fourth pathogen, Yersinia pestis, which is the etiologic agent of plague. To date, human lice belonging to the genus Pediculus have been classified into three mitochondrial clades: A, B, and C. Here, we describe a fourth mitochondrial clade, Clade D, comprising head and body lice. Clade D may be a vector of B. quintana and Y. pestis, which is prevalent in a highly plague-endemic area near the Rethy Health District, Orientale Province, Democratic Republic of the Congo.

Studies of Ancient Lice Reveal Unsuspected Past Migrations of Vectors.

Lice are among the oldest parasites of humans representing an excellent marker of the evolution and migration of our species over time. Here, we analyzed by real-time polymerase chain reaction (RT-PCR) developed in this study the mitochondrial DNA of seven ancient head louse eggs found on hair remains recovered from two sites in Israel: 1) five nits dating from Chalcolithic period (4,000 bc) were found in the Cave of the Treasure located at Nahal Mishmar, in the Judean Desert and 2) two nits dating from Early Islamic Period (ad 650-810) were found in Nahal Omer in the Arava Valley (between Dead Sea and Red Sea). Our results suggest that these eggs belonged to people originating from west Africa based on identification of the louse mitochondrial sub-clade specific to that region.

Detection of Bartonella quintana in African body and head lice.

Currently, the body louse is the only recognized vector of Bartonella quintana, an organism that causes trench fever. In this work, we investigated the prevalence of this bacterium in human lice in different African countries. We tested 616 head lice and 424 body lice from nine African countries using real-time polymerase chain reaction targeting intergenic spacer region 2 and specific B. quintana genes. Overall, B. quintana DNA was found in 54% and 2% of body and head lice, respectively. Our results also show that there are more body lice positive for B. quintana in poor countries, which was determined by the gross domestic product, than in wealthy areas (228/403 versus 0/21, P < 0.001). A similar finding was obtained for head lice (8/226 versus 2/390, P = 0.007). Our findings suggest that head lice in Africa may be infected by B. quintana when patients live in poor economic conditions and are also exposed to body lice.

Effect of permethrin-impregnated underwear on body lice in sheltered homeless persons: a randomized controlled trial.

IMPORTANCE: The control of body lice in homeless persons remains a challenge. OBJECTIVE: To determine whether the use of long-lasting insecticide-treated underwear provides effective long-term protection against body lice in homeless persons. DESIGN, SETTING, AND PARTICIPANTS: A randomized, double-blind, placebo-controlled trial was conducted in February and December 2011 in 2 homeless shelters (Madrague Ville and Forbin) in Marseille, France. Of the 125 homeless persons screened for eligibility, 73 body lice-infested homeless persons, 18 years or older, were enrolled. INTERVENTIONS: Body lice-infested homeless persons were randomly assigned to receive 0.4% permethrin-impregnated underwear or an identical-appearing placebo for 45 days, in a 1:1 ratio, with a permuted block size of 10. Visits were scheduled at days 14 and 45. Data regarding the presence or absence of live body lice were collected. MAIN OUTCOMES AND MEASURES: The primary and secondary end points were the proportions of homeless persons free of body lice on days 14 and 45, respectively. Mutations associated with permethrin resistance in the body lice were also identified. RESULTS: Significantly more homeless persons receiving permethrin-impregnated underwear than homeless persons receiving the placebo were free of body lice on day 14 in the intent-to-treat population (28% vs 9%; P = .04), with a between-group difference of 18.4 percentage points (95% CI, 1.4-35.4), and in the per-protocol population (34% vs 11%; P = .03), with a between-group difference of 23.7 percentage points (95% CI, 3.6-43.7). This difference was not sustained on day 45. At baseline, the prevalence of the permethrin-resistant haplotype was 51% in the permethrin group and 44% in the placebo group. On day 45, the permethrin-resistant haplotype was significantly more frequent in the permethrin group than in the placebo group (73% vs 45%, P < .001). CONCLUSION AND RELEVANCE: Permethrin-impregnated underwear is more efficient than placebo at eliminating body louse infestations by day 14; however, this difference was not sustained on day 45. The use of permethrin may have increased the resistance to permethrin in body lice and thus must be avoided. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT01287663.

Evidence of sympatry of clade a and clade B head lice in a pre-Columbian Chilean mummy from Camarones.

Three different lineages of head lice are known to parasitize humans. Clade A, which is currently worldwide in distribution, was previously demonstrated to be present in the Americas before the time of Columbus. The two other types of head lice are geographically restricted to America and Australia for clade B and to Africa and Asia for clade C. In this study, we tested two operculated nits from a 4,000-year-old Chilean mummy of Camarones for the presence of the partial Cytb mitochondrial gene (270 bp). Our finding shows that clade B head lice were present in America before the arrival of the European colonists.

Distinguishing body lice from head lice by multiplex real-time PCR analysis of the Phum_PHUM540560 gene.

BACKGROUND: Body louse or head louse? Once removed from their environment, body and head lice are indistinguishable. Neither the morphological criteria used since the mid-18th century nor the various genetic studies conducted since the advent of molecular biology tools have allowed body lice and head lice to be differentiated. In this work, using a portion of the Phum_PHUM540560 gene from the body louse, we aimed to develop a multiplex real-time polymerase chain reaction (PCR) assay to differentiate between body and head lice in a single reaction. MATERIALS AND METHODS: A total of 142 human lice were collected from mono-infested hosts from 13 countries on five continents. We first identified the louse clade using a cytochrome b (CYTB) PCR sequence alignment. We then aligned a fragment of the Phum_PHUM540560 gene amplified from head and body lice to design-specific TaqMan(©) FAM- and VIC-labeled probes. RESULTS: All the analyzed lice were Clade A lice. A total of 22 polymorphisms between the body and head lice were characterized. The multiplex real-time PCR analysis enabled the body and head lice to be distinguished in two hours. This method is simple, with 100% specificity and sensitivity. CONCLUSIONS: We confirmed that the Phum_PHUM540560 gene is a useful genetic marker for the study of lice.

Matrix-assisted laser desorption ionization-time of flight mass spectrometry for rapid identification of tick vectors.

A method for rapid species identification of ticks may help clinicians predict the disease outcomes of patients with tick bites and may inform the decision as to whether to administer postexposure prophylactic antibiotic treatment. We aimed to establish a matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) spectrum database based on the analysis of the legs of six tick vectors: Amblyomma variegatum, Rhipicephalus sanguineus, Hyalomma marginatum rufipes, Ixodes ricinus, Dermacentor marginatus, and Dermacentor reticulatus. A blind test was performed on a trial set of ticks to identify specimens of each species. Subsequently, we used MALDI-TOF MS to identify ticks obtained from the wild or removed from patients. The latter tick samples were also identified by 12S ribosomal DNA (rDNA) sequencing and were tested for bacterial infections. Ticks obtained from the wild or removed from patients (R. sanguineus, I. ricinus, and D. marginatus) were accurately identified using MALDI-TOF MS, with the exception of those ticks for which no spectra were available in the database. Furthermore, one damaged specimen was correctly identified as I. ricinus, a vector of Lyme disease, using MALDI-TOF MS only. Six of the 14 ticks removed from patients were found to be infected by pathogens that included Rickettsia, Anaplasma, and Borrelia spp. MALDI-TOF MS appears to be an effective tool for the rapid identification of tick vectors that requires no previous expertise in tick identification. The benefits for clinicians include the more targeted surveillance of patients for symptoms of potentially transmitted diseases and the ability to make more informed decisions as to whether to administer postexposure prophylactic treatment.

Detection of a knockdown resistance mutation associated with permethrin resistance in the body louse Pediculus humanus corporis by use of melting curve analysis genotyping.

Louse-borne diseases are prevalent in the homeless, and body louse eradication has thus far been unsuccessful in this population. We aim to develop a rapid and robust genotyping method usable in large field-based clinical studies to monitor permethrin resistance in the human body louse Pediculus humanus corporis. We assessed a melting curve analysis genotyping method based on real-time PCR using hybridization probes to detect the M815I-T917I-L920F knockdown resistance (kdr) mutation in the paraorthologous voltage-sensitive sodium channel (VSSC) α subunit gene, which is associated with permethrin resistance. The 908-bp DNA fragment of the VSSC gene, encoding the α subunit of the sodium channel and encompassing the three mutation sites, was PCR sequenced from 65 lice collected from a homeless population. We noted a high prevalence of the 3 indicated mutations in the body lice collected from homeless people (100% for the M815I and L920F mutations and 56.73% for the T917I mutation). These results were confirmed by melting curve analysis genotyping, which had a calculated sensitivity of 100% for the M815I and T917I mutations and of 98% for the L920F mutation. The specificity was 100% for M815I and L920F and 96% for T917I. Melting curve analysis genotyping is a fast, sensitive, and specific tool that is fully compatible with the analysis of a large number of samples in epidemiological surveys, allowing the simultaneous genotyping of 96 samples in just over an hour (75 min). Thus, it is perfectly suited for the epidemiological monitoring of permethrin resistance in human body lice in large-scale clinical studies.

Characterization of the novel Listeria monocytogenes PCR serogrouping profile IVb-v1.

The World Health Organization Collaborating Centre for Listeria (WHOCCL) has developed in 2004 a multiplex PCR assay that separates the 4 major Listeria monocytogenes serovars (1/2a, 1/2b, 1/2c, and 4b) into distinct PCR serogroups. A new PCR profile has been recently identified, constituted of amplified DNA fragments of prs, ORF2819, ORF2110 and lmo0737. Here we characterize 22 L. monocytogenes isolates of the WHOCCL collection with this PCR IVb variant 1 (IVb-v1) profile. The 22 isolates belong to the clinically predominant serovar 4b, exhibit 6 distinct pulsed-field gel electrophoresis ApaI/AscI combined profiles, and belong to 2 unrelated multilocus sequence types, indicating that the novel profile does not correspond to a recent clonal emergence. We have updated the WHOCCL serogroup-related PCR typing scheme to include this new profile.