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Human lice, Pediculus humanus, are obligate blood-sucking parasites. Body lice, Pediculus h. humanus, occur in two divergent mitochondrial clades (A and D) each exhibiting a particular geographic distribution. Currently, the body louse is recognized as the only vector for louse-borne diseases. In this study, we aimed to study the genetic diversity of body lice collected from homeless populations in three localities of northern Algeria, and to investigate louse-borne pathogens in these lice. In this study, 524 body lice specimens were collected from 44 homeless people in three localities: Algiers, Tizi Ouzou and Boumerdès located in northern Algeria. Duplex clade specific real-time PCRs (qPCR) and Cytochrome b (cytb) mitochondrial DNA (mtDNA) analysis were performed in order to identify the mitochondrial clade. Screening of louse-borne pathogens bacteria was based on targeting specific genes for each pathogen using qPCR supplemented by sequencing. All body lice belong to clade A. Through amplification and sequencing of the cytb gene we confirmed the presence of three haplotypes: A5, A9 and A63, which is novel. The molecular investigation of the 524 body lice samples revealed the presence of four human pathogens: Bartonella quintana (13.35%), Coxiella burnetii (10.52%), Anaplasma phagocytophilum (0.76%) and Acinetobacter species (A. baumannii, A. johnsonii, A. berezeniae, A. nosocomialis and A. variabilis, in total 46.94%). To the best of our knowledge, our study is the first to show the genetic diversity and presence of several emerging pathogenic bacteria in homeless' body lice from Algeria. We also report for the first time, the presence of several species of Acinetobacter in human body lice. Our results highlight the fact that body lice may be suspected as being a much broader vector of several pathogenic agents than previously thought. Nevertheless, other studies are needed to encourage epidemiological investigations and surveys of louse-associated infections.

Human lice have long been parasites of humans and are responsible for several epidemics in human medical history. However, their morphology, particularly head and body lice, remains very similar. In this study, we discriminate human louse species based on the number, shape and size of aeropyles (respiratory pores) of the egg operculum as well as the size of the respiratory orifice in the aeropyles. For this purpose, lice and eggs were collected from infested patients referring to Parasitology Department of Avicenne Hospital (Bobigny, France) and analyzed morphologically. The microscopic examination of the aeropyles revealed a large variation in the number ranging from 5 to 12 for head lice, 10 to 16 for body lice and 13 to 18 for pubic lice with a statistically distinct average of 8.3, 12.9 and 15.7, respectively. Although the length of the head and body louse eggs were almost similar, the width of body louse eggs were bigger than those of head louse (412.9 μm and 363.5 μm, respectively), while pubic louse eggs had the smallest length (692.9 μm) and width (286.1 μm). In addition, pubic louse eggs had a greater operculum length and width (253.7 μm length and 220.7 μm width) than head (230.6 μm length and 210.2 μm width) and body lice (227.9 μm length and 199.9 μm width). Moreover, pubic lice had the smallest average aeropyle size (41.2 μm) compared to head (51.1 μm) and body lice (49.1 μm). The respiratory orifice on the aeropyles of head and body lice (9.9 μm) were almost twice as big as those of pubic lice (4.9 μm). There was an inverse correlation between the number of aeropyles and their size in head, body and pubic lice. These findings are helpful in accurate identification of human louse species using their eggs, which could be useful for control management strategies against these ectoparasites.

  Selected morphological and biological differences between human head and body lice. Because of somewhat effective treatment options and increased societal standards for clothing and body hygiene, body lice are currently quite rare in most developed countries [9]. The number of homeless persons has increased significantly in recent decades, and the medical welfare of these people can be difficult to monitor for various reasons [10]. Since homeless persons may not have a change of clothing or be able to adequately delouse their clothes, their garb provides nourishing and unique environments needed for deposition and maintenance of body louse eggs [9], [10].

The human head louse, Pediculus humanus capitis, is subdivided into several significantly divergent mitochondrial haplogroups, each with particular geographical distributions. Historically, they are among the oldest human parasites, representing an excellent marker for tracking older events in human evolutionary history. In this study, ancient DNA analysis using real-time polymerase chain reaction (qPCR), combined with conventional PCR, was applied to the remains of twenty-four ancient head lice and their eggs from the Roman period which were recovered from Israel. The lice and eggs were found in three combs, one of which was recovered from archaeological excavations in the Hatzeva area of the Judean desert, and two of which found in Moa, in the Arava region, close to the Dead Sea. Results show that the head lice remains dating approximately to 2,000 years old have a cytb haplogroup A, which is worldwide in distribution, and haplogroup B, which has thus far only been found in contemporary lice from America, Europe, Australia and, most recently, Africa. More specifically, this haplogroup B has a B36 haplotype, the most common among B haplogroups, and has been present in America for at least 4,000 years. The present findings confirm that clade B lice existed, at least in the Middle East, prior to contacts between Native Americans and Europeans. These results support a Middle Eastern origin for clade B followed by its introduction into the New World with the early peoples. Lastly, the presence of Acinetobacter baumannii DNA was demonstrated by qPCR and sequencing in four head lice remains belonging to clade A.

Head lice, Pediculus humanus capitis, occur in four divergent mitochondrial clades (A, B, C and D), each having particular geographical distributions. Recent studies suggest that head lice, as is the case of body lice, can act as a vector for louse-borne diseases. Therefore, understanding the genetic diversity of lice worldwide is of critical importance to our understanding of the risk of louse-borne diseases. Here, we report the results of the first molecular screening of pygmies' head lice in the Republic of Congo for seven pathogens and an analysis of lice mitochondrial clades. We developed two duplex clade-specific real-time PCRs and identified three major mitochondrial clades: A, C, and D indicating high diversity among the head lice studied. We identified the presence of a dangerous human pathogen, Borrelia recurrentis, the causative agent of relapsing fever, in ten clade A head lice, which was not reported in the Republic of Congo, and B. theileri in one head louse. The results also show widespread infection among head lice with several species of Acinetobacter. A. junii was the most prevalent, followed by A. ursingii, A. baumannii, A. johnsonii, A. schindleri, A. lwoffii, A. nosocomialis and A. towneri. Our study is the first to show the presence of B. recurrentis in African pygmies' head lice in the Republic of Congo. This study is also the first to report the presence of DNAs of B. theileri and several species of Acinetobacter in human head lice. Further studies are needed to determine whether the head lice can transmit these pathogenic bacteria from person to another.

Background Head lice, Pediculus humanus capitis , are obligate blood-sucking parasites. Phylogenetically, they occur in five divergent mitochondrial clades (A, D, B, C and E), each having a particular geographical distribution. Recent studies have revealed that head lice, as is the case of body lice, can act as a vector for louse-borne diseases. Here, we aimed to study the genetic diversity of head lice collected from Niger’s refugees (migrant population) arriving in Algeria, northern Africa, and to look for louse-borne pathogens. Comparative head lice samples collected from indigenous population of schoolchildren (non-immigrant) were also analyzed to frame the study. Results In this study, 37 head lice samples were collected from 31 Nigerien refugees, as well as 45 head lice from 27 schoolchildren. The collection was established in three localities of eastern Algiers, north Algeria. Quantitative real-time PCR screening of pathogens bacteria and the genetic characterisation of the head lice satut were performed. Through amplification and sequencing of the cytb gene, results showed that all head lice of Nigerien refugees 37/82 (45.12%) belonged to clade E with the presence of four new haplotypes, while, of the 45 head lice of schoolchildren, 34/82 lice (41.46%) belonged to clade A and 11/82 (13.41%) belonged to clade B. Our study is the first to report the existence of clade E haplogroup in Nigerien head lice. DNA of Coxiella burnetii was detected in 3/37 (8.10%) of the head lice collected from 3 of the 31 (9.67%) migrant population. We also revealed the presence of Acinetobacter DNA in 20/37 (54.05%) of head lice collected from 25/31 (80.64%) of the Nigerien refugees, and in 25/45 (55.55%) head lice collected from 15/27 (55.55%) schoolchildren. All positive Nigerien-head lice for Acinetobacter spp. were identified as A. baumannii , while positive schoolchildren-head lice were identified as A. johnsonii 15/25 (60%), A. variabilis 8/25 (32%) and A. baumannii 2/25 (8%). Conclusions Based on these findings from head lice collected on migrant and non-migrant population, our results show, for the first time, that head lice from Niger belong to haplogroup E, and confirm that the clade E had a west African distribution. We also detected, for the first time, the presence of C. burnetii and A. baumannii in these Nigerien head lice. Nevertheless, further studies are needed to determine whether the head lice can transmit these pathogenic bacteria from one person to another.

Background Head lice ( Pediculus humanus capitis ) infestation is a worldwide public health concern, especially in school-aged children. However, its main impact factors and genetic characteristics remain poorly understood in China. Hence, the purpose of the study was to explore the precise association between multiple impact factors and head lice infestation, characterize the genetic variation of the head lice, and develop a sensitive and specific mitochondrial (mt) DNA duplex polymerase chain reaction (PCR) for accurately distinguishing clades A and B. Methods A cross-sectional study was conducted in Hunan Province, central China from January 2018 to July 2023. A total of 9254 schoolchildren from 48 primary schools in each administrative region were examined for head lice. Impact factors for infestation were analyzed using the data collected by a questionnaire. The mt cytb gene sequences of head lice collected in the current study were used for sequence analysis, then were added to the global pool to conduct the phylogenetic analyses. Primers designed on the basis of this gene sequence were used in duplex PCR to diagnose head lice clades A and B by amplicon size. Results Head lice infestation was found in 93.8% (45/48) of the primary schools included in the study. Overall, 6.8% (630/9254) of the examined schoolchildren harbored head lice, with 94.6% (596/630) being girls. A total of 2132 adult head lice were collected from 630 infested cases. The impact factors for head lice infestation included gender, school location, family situation, per capita income, study mode, and hair washing per week ( p  < 0.01). However, season and age were not considered as impact factors for head lice prevalence ( p  > 0.05). Phylogenetic analysis based on mt cytb gene sequences showed that head lice are classified into two clades (A and B), with clade B being more dominant in Hunan Province, central China. The newly developed duplex PCR was able to differentiate clades A from B in China with 100% sensitivity and specificity. Conclusions Our findings revealed that head lice infestation is mostly associated with poverty and poor hygiene in Hunan Province, central China. It is crucial to consider the simultaneous surveillance of head lice infestation in schoolchildren in regions with low level of socioeconomic status; however, datasets from other provinces are warranted to confirm the findings. It further showed that clades A and B are common in central China and that the latter has emerged and become the dominant one. Graphic Abstract

The study analyzes sequence variation of two mitochondrial genes (COI, cytb) in Pediculus humanus from three countries (Egypt, Pakistan, South Africa) that have received little prior attention and integrates these results with prior data. Analysis indicates a maximum K2P distance of 10.3% among 960 COI sequences and 13.8% among 479 cytb sequences. Three analytical methods (BIN, PTP, ABGD) reveal five concordant OTUs for COI and cytb. Neighbor-Joining analysis of the COI sequences confirm five clusters; three corresponding to previously recognized mitochondrial clades A, B, C and two new clades, “D” and “E”, showing 2.3% and 2.8% divergence from their nearest neighbors (NN). Cytb data corroborate five clusters showing that clades “D” and “E” are both 4.6% divergent from their respective NN clades. Phylogenetic analysis supports the monophyly of all clusters recovered by NJ analysis. Divergence time estimates suggest that the earliest split of P. humanus clades occured slightly more than one million years ago (MYa) and the latest about 0.3 MYa. Sequence divergences in COI and cytb among the five clades of P. humanus are 10X those in their human host, a difference that likely reflects both rate acceleration and the acquisition of lice clades from several archaic hominid lineages.

Background The head louse, Pediculus humanus capitis , is the most important ectoparasite causing many health problems. Several linkages are presented for this parasite, each representing a particular geographical distribution, prevalence rate, vector competence, susceptibility to pediculicides, and infestation rate. Determining the genetic nature of these linkages is necessary to identify the population structure and also to develop and monitor control programmes against head lice. This study was designed to analyse cox 1 and cytb genes and determine the mitochondrial clades in head lice populations in the northwest of Iran. Methods Adult head lice were collected from infested females of Ardabil, East and West Azerbaijan, and Zanjan Provinces from 2016 to 2018. Partial fragments of the mitochondrial genes cox 1 and cytb were amplified by PCR and some of the amplicons were sequenced. All confirmed sequences were analysed, and the frequency of each mitochondrial clade was determined in the studied areas. Results A total of 6410 females were clinically examined, and 897 adult head lice were collected from 562 infested cases. Genomic DNA was extracted from 417 samples, and fragments of cox 1 and c ytb genes were amplified in 348 individuals. Analysis of the 116 sequences showed the 632-bp and 495-bp fragments for cox 1 and cytb genes, respectively. The nucleotide and haplotype diversities of cytb and cox 1 genes were 0.02261 and 0.589 and 0.01443 and 0.424, respectively. Sequence analysis indicated 6 haplotypes clustered in two clades, A and B. The relative prevalence of clade B was 73% for cytb and 82% for cox 1 gene. Haplotypes of clade B were found in all the studied areas, while those of clade A were observed only in rural and suburban areas. Conclusions To our knowledge, this is the first study investigated deeply the field populations of Pediculus and documented two clades in the Middle East. The considerable prevalence of pediculosis in the studied areas requires authorities’ attention to establish effective control and preventive measures. Given the role of cytb in monitoring population groups, application of this marker is suggested for future epigenetic studies to evaluate the factors affecting the abundance of these clades.

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. 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. 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. We confirmed that the Phum_PHUM540560 gene is a useful genetic marker for the study of lice.