Determination of Knockdown Resistance (kdr) Allele Frequencies (T929I mutation) in Head Louse Populations from Mexico,Canada, and Peru

The head louse Pediculus humanus capitis (De Geer) is a hematophagous ectoparasite that inhabits the human scalp. The infestations are asymptomatic; however, skin irritation from scratching occasionally may cause secondary bacterial infections. The present study determined the presence and frequency of the knockdown resistance (kdr) mutation T929I in 245 head lice collected from Mexico, Peru, and Canada. Head lice were collected manually using a comb in the private head lice control clinic. Allele mutation at T9291 was present in 100% of the total sampled populations (245 lice) examined. In addition, 4.89% of the lice were homozygous susceptible, whereas 6.93% heterozygous and 88.16% homozygous were resistant, respectively. This represents the second report in Mexico and Quebec and fist in Lima.     

The mitochondrial genome of the chimpanzee louse,Pediculus schaeffi: insights into the process of mitochondrial genome fragmentation in the blood-sucking lice of great apes

Background

Blood-sucking lice in the genera Pediculus and Pthirus are obligate ectoparasites of great apes. Unlike most bilateral animals, which have 37 mitochondrial (mt) genes on a single circular chromosome, the sucking lice of humans have extensively fragmented mt genomes. The head louse, Pediculus capitis, and the body louse, Pe. humanus, have their 37 mt genes on 20 minichromosomes. The pubic louse, Pthirus pubis, has its 34 mt genes known on 14 minichromosomes. To understand the process of mt genome fragmentation in the sucking lice of great apes, we sequenced the mt genome of the chimpanzee louse, Pe. schaeffi, and compared it with the three human lice.

Results

We identified all of the 37 mt genes typical of bilateral animals in the chimpanzee louse; these genes are on 18 types of minichromosomes. Seventeen of the 18 minichromosomes of the chimpanzee louse have the same gene content and gene arrangement as their counterparts in the human head louse and the human body louse. However, five genes, cob, trnS₁, trnN, trnE and trnM, which are on three minichromosomes in the human head louse and the human body louse, are together on one minichromosome in the chimpanzee louse.

Conclusions

Using the human pubic louse, Pt. pubis, as an outgroup for comparison, we infer that a single minichromosome has fragmented into three in the lineage leading to the human head louse and the human body louse since this lineage diverged from the chimpanzee louse ~6 million years ago. Our results provide insights into the process of mt genome fragmentation in the sucking lice in a relatively fine evolutionary scale.

Electronic supplementary material

The online version of this article (doi:10.1186/s12864-015-1843-3) contains supplementary material, which is available to authorized users.     

Human lice show photopositive behaviour to white light

We studied the behavioural response of body lice and head lice to white light. We also evaluated the influence of starvation and the presence of other individuals on this response. Experiments were performed in a rectangular arena, half of which was illuminated and the other half kept in the dark. Two experiments were performed: in the first, a single louse was released into the arena for 60 min and the percentage of time spent in the illuminated half was recorded; in the second experiment, a group of lice was released and the number of insects in the illuminated half was recorded. The results showed that the average number of lice and time spent in the illuminated side of the arena was statistically higher than for the controls. Starvation did not influence the reaction of lice, but the number of insects in the illuminated area did increase with the size of the group. This study shows that human lice are photopositive towards white light and that this behaviour is not affected by the nutritional state of the insects. Moreover, it is enhanced by the presence of other lice.     

Immunization of rabbits with a midgut extract of the human body louse Pediculus humanus humanus: the effect of induced resistance on the louse population

Abstract. Resistance to human body lice, Pediculus humanus humanus L, induced by feeding on rabbits immunized with an extract of louse gut was studied. The mortality of lice fed on immunized rabbits was 73%, significantly higher than that of lice fed on control rabbits (52%) ( P < 0.01). The proportion of dead nymphs and female lice with ruptured guts was significantly higher in lice fed on immunized rabbits (P<0.01). The size of the bloodmeal was 35% greater in female lice fed on control rabbits than on immunized rabbits. Lice fed on immunized rabbits laid 40% less eggs than those fed on the controls, they also demonstrated a significant decrease in the number of eggs per female over time ( P < 0.01). 86% of the eggs laid by lice fed on immunized animals hatched, compared with 92% hatching of eggs laid by the lice fed on control animals (P< 0.01). With the exception of the first bloodmeal the percentage of hatched eggs which were laid between any two bloodmeals was significantly smaller (p<0.01) in the lice fed on immunized rabbits than in the control group. The first nymphal stage of lice fed on immunized rabbits took an average of 5.2 days to moult to the second stage, compared with 4 days for those fed on control rabbits.     

Genotyping of Human Lice Suggests Multiple Emergences of Body Lice from Local Head Louse Populations

Background

Genetic analyses of human lice have shown that the current taxonomic classification of head lice (Pediculus humanus capitis) and body lice (Pediculus humanus humanus) does not reflect their phylogenetic organization. Three phylotypes of head lice A, B and C exist but body lice have been observed only in phylotype A. Head and body lice have different behaviours and only the latter have been involved in outbreaks of infectious diseases including epidemic typhus, trench fever and louse borne recurrent fever. Recent studies suggest that body lice arose several times from head louse populations.

Methods and Findings

By introducing a new genotyping technique, sequencing variable intergenic spacers which were selected from louse genomic sequence, we were able to evaluate the genotypic distribution of 207 human lice. Sequence variation of two intergenic spacers, S2 and S5, discriminated the 207 lice into 148 genotypes and sequence variation of another two intergenic spacers, PM1 and PM2, discriminated 174 lice into 77 genotypes. Concatenation of the four intergenic spacers discriminated a panel of 97 lice into 96 genotypes. These intergenic spacer sequence types were relatively specific geographically, and enabled us to identify two clusters in France, one cluster in Central Africa (where a large body louse outbreak has been observed) and one cluster in Russia. Interestingly, head and body lice were not genetically differentiated.

Conclusions

We propose a hypothesis for the emergence of body lice, and suggest that humans with both low hygiene and head louse infestations provide an opportunity for head louse variants, able to ingest a larger blood meal (a required characteristic of body lice), to colonize clothing. If this hypothesis is ultimately supported, it would help to explain why poor human hygiene often coincides with outbreaks of body lice. Additionally, if head lice act as a reservoir for body lice, and that any social degradation in human populations may allow the formation of new populations of body lice, then head louse populations are potentially a greater threat to humans than previously assumed.     

Engorgement response of human body lice Pediculus humanus (Insecta: Anoplura) to blood fractions and their components

ABSTRACT. Fed through a synthetic membrane, 80% of females of the human body louse, Pediculus humanus de Geer, engorged on whole human blood compared to 30% on platelet poor plasma. Both the plasma albumin and small molecular weight components of the cellular fraction seemed to stimulate engorgement by lice. Known haemo-phagostimulants such as adenosine triphosphate and diphosphoglyceric acid when added to plasma, did not replace the small molecular weight components of the cellular fraction as feeding stimuli. Nymphs discriminated less than adults in selecting blood fractions.     

The digestive system of human lice: current advances and potential applications

Abstract. Haematophagy has evolved independently in different insect groups. This has resulted in the development of different kinds of adaptations (e.g. suitable mouthparts as well as alterations in the gut anatomy and in digestive enzyme composition). At the same time, insects have acquired different levels of adaptation to blood‐sucking habits. One of the most interesting examples is the co‐evolution of sucking lice (Anoplura) with their hosts. Human head and body lice, Pediculus humanus capitis De Geer, 1767 and Pediculus humanus humanus Linnaeus, 1758, are highly host‐ and anatomic‐region‐specific ectoparasites. They spend their entire lives on the host, ingesting small blood meals, several times per day. To date, several genes encoding digestive protease and proteolytic activities have been identified. The blood is stored in the anterior distensible region of the midgut, where genes encoding serine endopeptidases are highly expressed. Likewise, in the narrow posterior region of the midgut, the high‐level expression of genes encoding exopeptidases has been documented. Symbiotic microorganisms inhabiting the mycetome complete the deficient nutrition of human lice. This review focuses on the digestion habits of the best‐investigated taxon of human lice P. h. humanus in relation to other blood‐sucking insects.     

Nuclear Genetic Diversity in Human Lice (Pediculus humanus) Reveals Continental Differences and High Inbreeding among Worldwide Populations

Understanding the evolution of parasites is important to both basic and applied evolutionary biology. Knowledge of the genetic structure of parasite populations is critical for our ability to predict how an infection can spread through a host population and for the design of effective control methods. However, very little is known about the genetic structure of most human parasites, including the human louse (Pediculus humanus). This species is composed of two ecotypes: the head louse (Pediculus humanus capitis De Geer), and the clothing (body) louse (Pediculus humanus humanus Linnaeus). Hundreds of millions of head louse infestations affect children every year, and this number is on the rise, in part because of increased resistance to insecticides. Clothing lice affect mostly homeless and refugee-camp populations and although they are less prevalent than head lice, the medical consequences are more severe because they vector deadly bacterial pathogens. In this study we present the first assessment of the genetic structure of human louse populations by analyzing the nuclear genetic variation at 15 newly developed microsatellite loci in 93 human lice from 11 sites in four world regions. Both ecotypes showed heterozygote deficits relative to Hardy–Weinberg equilibrium and high inbreeding values, an expected pattern given their parasitic life history. Bayesian clustering analyses assigned lice to four distinct genetic clusters that were geographically structured. The low levels of gene flow among louse populations suggested that the evolution of insecticide resistance in lice would most likely be affected by local selection pressures, underscoring the importance of tailoring control strategies to population-specific genetic makeup and evolutionary history. Our panel of microsatellite markers provides powerful data to investigate not only ecological and evolutionary processes in lice, but also those in their human hosts because of the long-term coevolutionary association between lice and humans.     

Isoenzymes of human lice: pediculus humanus and P. capitis

Human lice (Phthiraptera: Pediculidae) from Africa, America and Europe were electrophoresed for 28 enzymes, with special interest in metabolic factors likely to be involved with insecticide resistance. Zymogram profiles of the body louse (Pediculus humanus L. from France and U.S.A.) and the head louse (P. capitis DeGeer from France, Madagascar, Mali & Senegal) were compared. Only esterase two enzymes, phosphoglucomutase (Pgm) and 3 (Est-3), showed electrophoretic variation. In our starch gel electrophoresis conditions, P. humanus showed three electromorphs of Pgm migrating anodally 6, 11 and 16 mm (designated alleles a, b, c, respectively). Of the putative Pgm alleles, b and c occurred in all samples of both species of lice, whereas allele a was found only in P. humanus lab strain from U.S.A. Esterase 3 had four electromorphs migrating 23, 26, 30 and 35 mm (designated alleles a, b, c and d). Among putative Est alleles, a was found only in P. capitis from Bamako (all 14 specimens aa homozygotes), allele d was found only in P. capitis from Dakar (39% frequency), whereas Est-3 alleles b and c showed apparently balanced polymorphism in all samples of both P. humanus and P. capitis except that from Bamako. Despite the limited amount of isoenzyme variation detected (only 2/31 polymorphic loci), divergences of Est-3 and Pgm among Pediculus populations may be relevant to their biosystematics and resistance.     

Permethrin resistance in the head louse Pediculus capitis from Israel

Head lice, Pediculus capitis , were collected from children aged 3–12 years in Maale Adumin, a town near Jerusalem, after reports of control failure with the pyrethroid insecticide permethrin. A total of 1516 children were examined: living lice and eggs were found on 12.1% of the children; or another 22.8% of the children only nits were found. Twice as many girls as boys (8.1% v 4%) were infested with lice and or nits. Head lice collected from infested children were exposed to permethrin impregnated filter-papers. Log time probit mortality (ltp) regression lines were calculated for mortality data and compared to ltp lines for a similar collection of head lice made in 1989. The regression lines for the two years were significantly different, with a 4-fold decrease in susceptibility at the LT50 level between 1989 and 1994. The slopes of the lines also suggested that the 1994 population was more heterogenous in its response to permethrin than the 1989 population. In contrast, a laboratory population of body lice (Pediculus humanus) tested with the same batch of permethrin-impregnated papers showed a slight but non-significant increase in susceptibility between 1989 and 1994. The results suggest that resistance to pyrethroids has developed rapidly among head lice since permethrin was introduced in 1991 as a pediculicide in Israel.     

Partial characterization and post-feeding activity of midgut aminopeptidase in the human body louse, Pediculus humanus humanus

Abstract. A leucine aminopeptidase was found in the midgut of the human body louse, Pediculus humanus humanus L. (Anoplura: Pediculidae). The enzyme is activated by the bloodmeal with a pH optimum at 8. The enzyme is soluble in both aqueous and detergent-containing solutions. The two forms of the enzyme had the same Km but exhibited different catalytic activities with regard to Vmax values in these solutions. The enzyme is inhibited competitively by a substrate analogue 1,10-phenanthroline and by Mn²⁺ ions in the presence and absence of detergent.     

Molecular evolution of Pediculus humanus and the origin of clothing

The human head louse (Pediculus humanus capitis) and body louse (P. humanus corporis or P. h. humanus) are strict, obligate human ectoparasites that differ mainly in their habitat on the host : the head louse lives and feeds exclusively on the scalp, whereas the body louse feeds on the body but lives in clothing. This ecological differentiation probably arose when humans adopted frequent use of clothing, an important event in human evolution for which there is no direct archaeological evidence. We therefore used a molecular clock approach to date the origin of body lice, assuming that this should correspond with the frequent use of clothing. Sequences were obtained from two mtDNA and two nuclear DNA segments from a global sample of 40 head and body lice, and from a chimpanzee louse to use as an outgroup. The results indicate greater diversity in African than non-African lice, suggesting an African origin of human lice. A molecular clock analysis indicates that body lice originated not more than about 72,000 +/- 42,000 years ago; the mtDNA sequences also indicate a demographic expansion of body lice that correlates with the spread of modern humans out of Africa. These results suggest that clothing was a surprisingly recent innovation in human evolution.     

What's in a name: the taxonomic status of human head and body lice

Human head lice (Anoplura: Pediculidae: Pediculus) are pandemic, parasitizing countless school children worldwide due to the evolution of insecticide resistance, and human body (clothing) lice are responsible for the deaths of millions as a result of vectoring several deadly bacterial pathogens. Despite the obvious impact these lice have had on their human hosts, it is unclear whether head and body lice represent two morphological forms of a single species or two distinct species. To assess the taxonomic status of head and body lice, we provide a synthesis of publicly available molecular data in GenBank, and we compare phylogenetic and population genetic methods using the most diverse geographic and molecular sampling presently available. Our analyses find reticulated networks, gene flow, and a lack of reciprocal monophyly, all of which indicate that head and body lice do not represent genetically distinct evolutionary units. Based on these findings, as well as inconsistencies of morphological, behavioral, and ecological variability between head and body lice, we contend that no known species concept would recognize these louse morphotypes as separate species. We recommend recognizing head and body lice as morphotypes of a single species, Pediculus humanus, until compelling new data and analyses (preferably analyses of fast evolving nuclear markers in a coalescent framework) indicate otherwise.     

Partial purification of the aminopeptidase from the midgut of the human body louse, Pediculus humanus humanus

The midgut of the human body louse Pediculus humanus humanus contains a thermally stable leucine aminopeptidase, which was detected by agarose gel electrophoresis using l ‐amino oxidase. Midgut extracts were homogenized in saline or in 1% Triton X‐100 and the aminopeptidase was purified by Superose 6 gel filtration chromatography. A peak with enzyme activity that was extracted with or without Triton X‐100 was eluted at a molecular weight 67–69 kDa. Non‐denaturing polyacrylamide gel electrophoresis resolved one band of molecular weight of 69 kDa for samples that were extracted in a saline buffer. Two closely linked bands of molecular weight 67 kDa and 69 kDa were observed in samples that were extracted in 1% Triton X‐100.     

Repellency of essential oils and their components to the human body louse, Pediculus humanus humanus

Five essential oils and nine of their components were compared to diethyl toluamide (DEET) for their repellent activity against the human body louse, Pediculus humanus humanus. The absolute or intrinsic repellency of the compounds was tested by applying the repellent to corduroy patches and comparing them with untreated patches. It was found that the most effective repellents were DEET and citronella, whose activity lasted at least 29 days. The activity of rosemary lasted at least 18 days and that of eucalyptus more than 8 days. The repellent activity of the oil components such as citronellal and geraniol lasted more than 15 and 8 days, respectively. DEET remained effective at a dilution of 1:32, geraniol at 1:8, citronella at 1:4 and rosemary and citronellal at 1:1. The comparative or standard repellency of the candidate repellents was examined with the aid of a new screening technique using hairs treated with ammonium bicarbonate which is attractive to lice. Using this technique it could be shown that the repellent activity of citronella and geraniol lasted 2 days and that of rosemary and citronellal for only one day. DEET was active for less than one day. Serial dilutions of these substances also revealed that citronella was the most potent repellent for lice, followed by citronellal, rosemary, geraniol and DEET. The differences however, were not significant.     

In Vitro Screening of Anti-lice Activity of Pongamia pinnata Leaves

Growing patterns of pediculocidal drug resistance towards head louse laid the foundation for research in exploring novel anti-lice agents from medicinal plants. In the present study, various extracts of Pongamia pinnata leaves were tested against the head louse Pediculus humanus capitis. A filter paper diffusion method was conducted for determining the potential pediculocidal and ovicidal activity of chloroform, petroleum ether, methanol, and water extracts of P. pinnata leaves. The findings revealed that petroleum ether extracts possess excellent anti-lice activity with values ranging between 50.3% and 100% where as chloroform and methanol extracts showed moderate pediculocidal effects. The chloroform and methanol extracts were also successful in inhibiting nymph emergence and the petroleum ether extract was the most effective with a complete inhibition of emergence. Water extract was devoid of both pediculocidal and ovicidal activities. All the results were well comparable with benzoyl benzoate (25% w/v). These results showed the prospect of using P. pinnata leave extracts against P. humanus capitis in difficult situations of emergence of resistance to synthetic anti-lice agents.     

Effects of blood type and blood handling on feeding success,longevity and egg production in the body louse,Pediculus humanus humanus

The effects of feeding different types of human blood to human body lice, Pediculus humanus humanus L. (Phthiraptera: Pediculidae), on feeding success, longevity and numbers of eggs laid were investigated using an artificial blood‐feeding system in the laboratory. No significant differences were found between lice fed on different human blood types for any of the parameters tested. However, when lice were fed on human blood of one blood type followed immediately by a different blood type, they took significantly smaller bloodmeals, their longevity was reduced and they laid fewer eggs per female than control lice that had been fed twice on the same human blood type. When lice were fed human blood that had been stored for 1–26 weeks, the quantity of blood taken, the proportion of lice that became fully engorged and lice longevity diminished gradually as the storage time of the blood increased, but there was no effect of storage time on the mean number of eggs laid per female. However, lice would not feed on 26‐week‐old blood. The type of anticoagulant used had a significant effect on the proportion fed, longevity and number of eggs laid per female. Generally, EDTA (ethylenediaminetetraacetic acid)‐treated blood reduced longevity and the number of eggs laid per female to a greater degree than heparinized or citrated blood. Lice fed on rabbit blood took significantly larger amounts of blood, lived longer and laid a higher mean number of eggs per female than lice fed on human blood.     

Immunization of rabbits with faecal extract of Pediculus humanus, the human body louse: effects on louse development and reproduction

Abstract Immunization of rabbits with a faecal extract of the human body louse (Pediculus humanus) induced a high titre of specific IgG. The mean weight of Wood taken by females fed on the immunized rabbits was significantly lower (29%) than taken by females fed on the control rabbits. The mean number of eggs per female fed on the immunized rabbits was significantly lower than for females fed on the control rabbits. The hatchability of the eggs laid by lice fed on immunized rabbits (91%) was significantly lower than of those fed on control rabbits (94%). The rate of development of nymphs fed on control rabbits was significantly higher than those fed on the immunized rabbits. There was no difference in survival rates of lice fed on immunized and control rabbits.     

Plant extracts and their components as potential control agents against human head lice

The head lice, Pediculus humanus capitis (Phthiraptera:Pediculidae), is an obligate ectoparasite of humans that causes pediculosis capitis, a nuisance for millions of people worldwide, with high prevalence in children. Pediculosis capitis has been treated by methods that include the physical remotion of lice, various domestic treatments and conventional insecticides. None of these methods render complete protection, and there is clear evidence for the evolution of resistance and cross-resistance to conventional insecticides. Non-toxic alternative options are hence needed for head lice treatment and/or prevention, and natural products from plants, especially essential oils (EOs), are good candidates for safer control agents that may provide good anti-lice activity and low levels of evolved resistance. A few EOs have been tested as repellents with promissory results, although often in vitro tests and clinical trials produce contradictory results. A handful of fixed extracts and several EOs and their individual components have also been tested as contact pediculicides or fumigants. The studies have focused mainly on plant families characterized for the production of EOs. While many EOs and individual compounds showed pediculicide activity, comparing results is difficult due to the diverse bioassay methodologies. Studies of anti-lice activity of individual EO components provide the basis for preliminary conclusions of structure–activity relationships, although no clear patterns can yet be drawn. We here attempt to provide a concise compilation of the available information on anti-lice activity of plant extracts and plant-derived compounds, which we hope may be of help for future developments in this area.

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