Pediculus humanus capitis: a study of the male genitalia using a combined stereoscopic, confocal laser scanning, and scanning electron microscopy approach.

BACKGROUND: The male genital structures of arthropods are key features in the taxonomic and phylogenetic study of these organisms. The male genitalia of the head louse Pediculus humanus capitis are complex organs which are partly composed of structures that dynamically extrude during copulation. METHODS: Here, we describe the morphology of the genitalia of P. humanus capitis at the copulation stage, and at rest, by using stereoscopic microscopy, confocal laser scanning microscopy (CLSM), and scanning electron microscopy (SEM). RESULTS: CLSM and SEM images revealed that the vesica is composed of two distinct anatomical parts, the proximal lobe and the distal lobe. Both lobes have short and narrow spines, as well as long and wide scales with either sharp or rounded tips. The rounded scales vary in size and have a wavy base and rounded tips, and thus resemble a tongue in appearance. We identified a gland-like area on the penis with 11 shallow circular depressions, and a flat area with 14-16 exit orifices. The apical end of the penis has a foliaceous trifurcation and serves to expel the contents of the ejaculatory duct. These characteristics were recorded for all the specimens analyzed, indicating that these structures are highly conserved; to our knowledge, they have not been previously reported for any suborder of lice. CONCLUSIONS: To the best of our knowledge, our results reveal for the first time the morphological details, and complexity, of the male genitalia of the head louse P. humanus capitis at different stages of copulation. The new approach described here provided information that should be taken into consideration in future research on the genitalia of lice. Application of this approach will also impact the taxonomic and phylogenetic study of other insect taxa.

Characterization of nit sheath protein functions and transglutaminase-mediated cross-linking in the human head louse, Pediculus humanus capitis.

BACKGROUND: Head louse females secrete liquid glue during oviposition, which is solidified to form the nit sheath over the egg. Recently, two homologous proteins, named louse nit sheath protein (LNSP) 1 and LNSP 2, were identified as adhesive proteins but the precise mechanism of nit sheath solidification is unknown. METHODS: We determined the temporal transcriptome profiles of the head louse accessory glands plus oviduct, from which putative major structural proteins and those with functional importance were deduced. A series of RNA interference (RNAi) experiments and treatment of an inhibitor were conducted to elucidate the function and action mechanism of each component. RESULTS: By transcriptome profiling of genes expressed in the louse accessory glands plus uterus, the LNSP1 and LNSP2 along with two hypothetical proteins were confirmed to be the major structural proteins. In addition, several proteins with functional importance, including transglutaminase (TG), defensin 1 and defensin 2, were identified. When LNSP1 was knocked down via RNA interference, most eggs became nonviable via desiccation, suggesting its role in desiccation resistance. Knockdown of LNSP2, however, resulted in oviposition failure, which suggests that LNSP2 may serve as the basic platform to form the nit sheath and may have an additional function of lubrication. Knockdown of TG also impaired egg hatching, demonstrating its role in the cross-linking of nit sheath proteins. The role of TG in cross-linking was further confirmed by injecting or hair coating of GGsTop, a TG inhibitor. CONCLUSIONS: Both LNSP1 and LNSP2 are essential for maintaining egg viability besides their function as glue. The TG-mediated cross-linking plays critical roles in water preservation that are essential for ensuring normal embryogenesis. TG-mediated cross-linking mechanism can be employed as a therapeutic target to control human louse eggs, and any topically applied TG inhibitors can be exploited as potential ovicidal agents.

Sensory features of the human louse antenna: new contributions and comparisons between ecotypes.

Body and head lice are known to be exclusive ectoparasites of human hosts. Current genomic and transcriptomic data suggest that both louse types represent ecotypes of the same species. They exhibit morphological and physiological differences that probably arose from living in different biotopes. Sensory traits represent suitable candidates to be affected by environmental heterogeneity. Therefore, through scanning electron microscope studies, this study analysed and compared the sensory structures of both ecotypes' antennae. Seven types of sensory structures were identified in both ecotypes: bristles, tuft organs, pore organs, single pore and three morphotypes of sensilla basiconica. Notably, the single pore and the morphotypes of sensilla basiconica were described for the first time in the body louse antenna. This study's comparative analysis mainly revealed size differences across the sensory structures of the ecotypes. Bristles of the flagellomere 2 of the body louse antenna were longer than the head louse bristles. In addition, the pore organs of the head louse antenna presented a higher diameter than those of the body louse. The possible relevance of size differences regarding the biotopes exploited by the body louse and the head louse is discussed. Yet, physiological studies may help to fully understand the phenotypical differences of both ecotypes.

Automontage microscopy and SEM: A combined approach for documenting ancient lice.

Human ectoparasites, including lice, have been recovered from a wide range of archaeological materials. The human head louse, Pediculus humanus capitis, has been identified from mummies and sediments for decades. Louse eggs are the body part most commonly encountered and therefore the most frequently quantified. Typically, several types of microscopy are applied for egg documentation. For studies in which quantification of infestation is a goal, counting is done with the naked eye or with the aid of handheld lenses. For determination and stage classification, stereomicroscopy is commonly used. For more detailed examination of microstructure, light microscopy, scanning electron microscopy (SEM), and confocal laser scanning microscopy (CLSM) can be employed. In most reports, researchers use two or more techniques to accomplish interrelated goals. Automontage microscopy is used to document prehistoric arthropods with good success. Herein, we report the results of a combination of SEM and automontage microscopy to document lice and eggs recovered from South American mummies. This combined approach allows for simultaneous examination of internal and external characteristics. Thirty automontage composite images of 2 adult lice and 16 eggs showed that egg internal morphologies were easily examined showing the within-egg anatomy of emergent nymphs. SEM imaging of 9 lice and 129 eggs was completed. In the case of two adults and several eggs, SEM imaging was accomplish after automontage image capture of the same specimens. This one-to-one image comparison of SEM and automontage shows that transmitted light of automontage reveals egg internal structures and details of the adult lice. SEM allows for high magnification examination of egg, nymph and adult microstructures. We conclude that automontage imaging followed by SEM results in efficient graphic documentation of rare louse specimens.

Diversity of mitochondrial genes and predominance of Clade B in different head lice populations in the northwest of Iran.

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 cox1 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 cox1 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 cox1 and cytb genes were amplified in 348 individuals. Analysis of the 116 sequences showed the 632-bp and 495-bp fragments for cox1 and cytb genes, respectively. The nucleotide and haplotype diversities of cytb and cox1 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 cox1 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.

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.

Prevalence of Bartonella quintana in patients with fever and head lice from rural areas of Sine-Saloum, Senegal.

Trench fever is poorly known by the staff of health facilities that manage febrile patients in Senegal. Bartonella quintana DNA was identified in 5 of 274 (2%) febrile patients from two rural dispensaries and 2 of 71 (3%) head lice specimens collected from the same villages.

Lice.

Pediculosis (capitis, corporis, and pubis) share well-known features: worldwide prevalence (involving millions of people annually); parasites inducing skin lesions directly, and indirectly as a result of itching and hypersensitivity to parasites; and treatment based on good entomological knowledge of the parasite and practical considerations (ie, most available treatments do not act on eggs and should be repeated, depending on the life cycle of the parasites). Infestations are spread most commonly by close contacts. Social stigma and persistent misconceptions complicate the implementation of appropriate management strategies. Head and pubic lice infestations are diagnosed by the visualization of insects or viable nits (eggs). Primary treatments are topical pediculicides (permethrin or malathion), used twice, but emergence of resistance against pediculicides has created the need of alternative treatments including topical or oral ivermectin. Pubic lice are treated the same as head lice, but this finding should prompt evaluation for other sexually transmitted diseases. Body lice infestation should be suspected when symptoms of generalized itching occur in persons who do not change or wash their clothing or bedding regularly; lice may be found in the seams of their clothing.Topically administered permethrin may help to eradicate body lice, but personal hygiene measures are essential for successful treatment. Environmental treatment is also necessary for the eradication of the infestation. Health care personnel who come into contact with this population need to be well informed of the facts in order to disseminate accurate information for diagnosis and management.

Biology and genetics of human head and body lice.

Head lice and body lice have distinct ecologies and differ slightly in morphology and biology, questioning their taxonomic status. Over the past 10 years many genetic studies have been undertaken. Controversial data suggest that not only body lice but also head lice can serve as vectors of Bartonella quintana, and a better understanding of louse epidemiology is crucial. Here, we review taxonomic studies based on biology and genetics, including genomic data on lice, lice endosymbionts, and louse-transmitted bacteria. We recommend that studies of human lice employ morphological and biological characteristics in conjunction with transcriptomic date because lice seem to differ mainly in gene expression (and not in gene content), leading to different phenotypes.

Head lice: scientific assessment of the nit sheath with clinical ramifications and therapeutic options.

Head lice, like many insects, produce a protective coating for their newly laid eggs that is essential to the survival of the species. Knowledge of the composition of the sheath, which is the glue by which the egg is attached to human hair, and the nit laying process could lead to production of agents that could be used to attack louse infestations by interfering with the normally protected environment of nymph development within the egg. The physical removal of nits has become an important part of treatment of head louse infestations given the "no-nit" policy in schools. Biochemical analysis has revealed that the nit sheath of the head louse is composed of 4 bands of protein, possibly cross-linked to aliphatic components with a tertiary structure of beta sheeting. Nature has protected the louse by making the nit sheath similar in composition to the hair; thereby, agents designed to unravel the nit sheath may also damage human hair. Possible targets to destroy the nit sheath include proteases, denaturants, beta sheet breaker proteins, and small protein inhibitors of sheath formation. Better understanding of insect glues may allow us to develop compounds so that the liquid secretions of the collateral glands of the female louse, which becomes the nit sheath, do not solidify by oxidation when placed with the louse egg onto human hair. Knowledge of insect behavior, such as oviposition, may also suggest methods for repelling female lice from laying eggs onto hair. Alternatively, agents that coat the nits and restrict the oxygen transfer to the developing larvae may prove beneficial.

Clinical update on resistance and treatment of Pediculosis capitis.

The anatomy and physiology of head lice make them extremely adaptable to their human hosts but also difficult to eradicate. Their coloring and small size make them difficult to see, and the larvae, or nymphs, have multiple exoskeletons for protection. When exposed to pediculicides, a nymph can shed its outer exoskeleton and receive only a sublethal dose. Through natural selection, head lice have developed resistance to commonly used pediculicides, making it increasingly difficult to eliminate infestations. Other contributors to resistance include changes in formulations of pediculicides and improper use. Over the last 20 years, the efficacy of pyrethrins has declined because of necessary safety changes in formulations and unintentional effects of new packaging on chemical components of these products. Pediculicides designed to be applied to wet hair may become too diluted to produce the desired effect. Patients also may use too little product to conserve costs. The combination of decreased product efficacy and exposure of head lice to diluted or insufficient amounts of pediculicides has reduced their effectiveness substantially. Studies comparing the current efficacy of several over-the-counter and prescription pediculicides with effectiveness in the 1980s has shown that OVIDE (malathion) Lotion, 0.5% is the only product that has retained its efficacy over time. The efficacy of over-the-counter pyrethrin products had declined significantly, and lindane, another prescription product, was least effective of all products tested both in the 1980s and in more recent studies. In addition to retaining its efficacy, malathion 0.5% also is effective when applied for only 20 minutes. The shorter application time is safer for children and also decreases the likelihood that residual product remains in the hair, further contributing to resistance.

Pediculosis.

The 3 major lice that infest humans are Pediculus humanus capitis (head louse), Pthirus pubis (crab louse), and Pediculus humanus humanus (body louse). Patients with louse infestation present with scalp pruritus, excoriations, cervical lymphadenopathy, and conjunctivitis. A hypersensitivity rash, or pediculid, may mimic a viral exanthem. Head lice infestation crosses all economic and social boundaries, whereas body lice infestation preferentially affects the homeless and displaced. Body lice are major vectors of diseases such as typhus, trench fever, and relapsing fever. Pubic lice infestation often is acquired as a sexually transmitted disease and may be a marker to screen for other sexually transmitted diseases. Treatment of louse infestation can be challenging. Mechanical measures, such as combing, are helpful as adjunctive measures, but most studies suggest they are not as effective as chemical agents. Resistance to chemical agents is a growing problem. Major types of resistance include knock-down resistance, glutathione-S-transferase-based resistance, and monooxygenase-based resistance. Research is needed to identify new effective treatments.

[The adaptive potentials of human head and clothes lice when parasitizing on man]. / Adaptatsionnye vozmozhnosti golovnykh i platianykh vshei cheloveka pri parazitirovanii na cheloveke.

Two species of lice (Pediculus capitis and P. corporis) from various areas of Russian and adjacent republics were examined and analysed for their specific features. Each species was characterized in terms of its morphology, karyology, biology, reproductive isolation, parasitic connections, epidemiology and role as casual organisms. The two species live apart on the same host and they can live on it simultaneously. P. corporis has a higher influence on the host's metabolism than P. capitis; P. corporis seems to be a younger species and its adaptation to man has not achieved the level that is inherent in P. capitis. The head lice (P. capitis) do not affect an epidemic process in typhus and so pediculosis should be identified by the species of lice.