In vitro efficacy of over-the-counter botanical pediculicides against the head louse Pediculus humanus var capitis based on a stringent standard for mortality assessment

Abstract.  Infestation of the head louse Pediculus humanus var capitis DeGeer (Phthiraptera: Pediculidae) is an important public health problem in Australia, with up to a third of children infested in some primary schools. Insecticide resistance and inadequate attention to the application instructions of topical pediculicides are common reasons for treatment failure. This study evaluated six popular Australian over-the-counter products against head lice, primarily comprised of different botanical extracts, and compared them with permethrin 1% (Quellada^®) and a non-treatment control in order to assess their in vitro efficacy. We also assessed commonly used criteria for evaluating pediculicide efficacy in vitro. All tested products failed to demonstrate high levels of efficacy with the exception of Tea Tree Gel^®, which outperformed 1% permethrin. Permethrin had a high level of efficacy, but using stringent criteria 18% of lice were not dead at 3 h, indicating some resistance to Quellada^®. Commonly used less stringent criteria were shown to overestimate mortality of head lice as a result of the protective phenomenon of stasis or sham death observed in exposed lice that may recover after some time. Using two different levels of stringency resulted in different rankings of efficacy for most products, with the exception of the first ranked product, Tea Tree Gel^®. Rankings of efficacy also varied over time, even within the different assessment criteria. Government regulatory agencies should require standard in vitro tests using stringent mortality criteria, with an observation period of ≥ 6 h, to determine the efficacy of new pediculicides, and only products that cause a minimum mortality rate (e.g. 80%) in head lice collected from the target population should be licensed for sale.     

Pyrethroid resistance mechanisms in the head louse Pediculus capitis from Israel: implications for control

In Israel, the head louse, Pediculus capitis, developed resistance to DDT through the extensive use of this insecticide until the 1980s. In 1991, permethrin was introduced for control of DDT resistant P. capitis in Israel, leading to control failure of this pyrethroid insecticide by 1994. Pyrethroid resistance of P. capitis in Israel extends to phenothrin, which has not been used for louse control. We identified a glutathione S-transferase(GST)-based mechanism of DDT resistance in the Israeli head lice. This GST mechanism occurred before 1989, while permethrin resistance in P. capitis developed after 1994, suggesting that the main GST resistance mechanism selected by DDT use does not confer any pyrethroid cross-resistance. Esterase activity levels were equivalent in pyrethroid resistant and susceptible P. capitis field-collected in Israel, and in a susceptible strain of P. humanus, the body louse, indicating no involvement of any esterase-based mechanism in resistance. A weak monooxygenase-based permethrin metabolism resistance mechanism was the only factor identified which could account for any of the observed pyrethroid resistance in P. capitis. However, the lack of synergism of phenothrin resistance by piperonyl butoxide suggests that a non-oxidative mechanism is also present in the resistant lice. Therefore it seems probable that pyrethroid resistance in Israeli P. capitis is due to a combination of nerve insensitivity (knockdown resistance or 'kdr') and monooxygenase resistance mechanisms.     

Infestation status of head louse and treatment with lindane shampoo in children of primary school and kindergarten in Chinju-shi, Kyongsangnam-do, Korea

The infestation status of head louse among children attending primary schools and kindergartens in Chinju-shi, Kyongsangnam-do, Korea, was investigated between June and July 1999. Out of 2,288 children examined, 3.9% of boys (48/1,242) and 23.5% of girls (246/1,046) were infested with nits or adult/nymphs of lice. The effectiveness of lindane shampoo (1% gamma benzene hexachloride solution) was evaluated after one or two time applications to all the children infested. The negative conversion rate of pediculosis was 93.5%. Effective control measures are needed to control and prevent such ectoparasite infestation amongst children.     

Prevalence of nits and lice in samples of cut hair from floors of barbershops and beauty parlors in Belo Horizonte, Minas Gerais State, Brazil

A louse survey based on samples of cut hair collected from floors of barbershops and beauty parlors was conducted in Belo Horizonte, Minas Gerais State, Brazil, from October 1984 to April 1985, as an alternative way to determine the prevalence of pediculosis capitis in the population. Of 475 samples examined for nits, nymphs, or adults of Pediculus capitis, 140 were infested (29.5%). A total of 58 lice and 3,553 nits were found in 33,632.9 g of hair collected, giving a ratio of 0.10 nit/g. Almost 29% of the nits were viable and capable of being transmitted after hatching. There was significant difference among the infestation rates by socioeconomic levels, and samples from barbershops with male customers were the most infested. Based upon the number of haircuts in each sample, we estimated that 5 or 6% of the population might be infested by this species.     

The prevalence of head louse infestation among urban and rural children in Korea

The prevalence of head louse (Pediculus hamanus var. capitis) infestation in preschool and school children in 9 urban and 8 rural areas of Korea, was investigated. Of 11,865 children who were examined for head lice and nits, 2,900(24.4%) were found infested. The over-all infestation rate of rural children, 58.9%, was higher than that of urban children, 14.4%. There also existed significant differences in the infestation rate among various regions of urban as well as rural areas. Females were more infested than males. But there was no differences in infestation rate in either sex of preschool age. The infestation rate increased gradually from the age of 6, reached a plateau between 9-12 years of age and thereafter slowly decreased. The infestation rate of an orphanage children was higher than that of general childhood population. Health education and mass-delousing attempt are urgently required to lower such a high prevalence of head louse infestation in Korea.     

Infestation of people with lice in Kathmandu and Pokhara, Nepal

The prevalence of infestation with head lice and body lice, Pediculus spp. (Phthiraptera: Pediculidae) and pubic (crab) lice Pthirus pubis (L.) (Phthiraptera: Pthiridae), was recorded from 484 people in Nepal. The prevalence of head lice varied from 16% in a sample of people aged 10-39 years of age, to 59% in street children. Simultaneous infestations with head and body lice (double infestations) varied from 18% in slum children to 59% in street children.     

Severe head lice infestation in an Andean mummy of Arica, Chile

Pediculus humanus capitis is an ancient human parasite, probably inherited from pre-hominid times. Infestation appears as a recurrent health problem throughout history, including in pre-Columbian populations. Here, we describe and discuss the occurrence of pre-Columbian pediculosis in the Andean region of the Atacama Desert. Using a light microscope and scanning electron microscopy, we studied a highly infested Maitas Chiribaya mummy from Arica in northern Chile dating to 670-990 calibrated years A.D. The scalp and hair of the mummy were almost completely covered by nits and adult head lice. Low- and high-vacuum scanning electron microscopy revealed a well-preserved morphology of the eggs. In addition, the excellent preservation of the nearly 1,000-yr-old adult head lice allowed us to observe and characterize the head, antennae, thorax, abdomen, and legs. Leg segmentation, abdominal spiracles, and sexual dimorphism also were clearly observed. The preservation of the ectoparasites allowed us to examine the micromorphology using scanning electron microscopy; the opercula, aeropyles, and spiracles were clearly visible. This case study provides strong evidence that head lice were a common nuisance for Andean farmers and herders. Head lice are transmitted by direct head-to-head contact; thus, this ancient farmer and herder was potentially infesting other people. The present study contributes to the body of research focusing on lice in ancient populations.     

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.     

Ovicidal and adulticidal effects of monoterpenoids against permethrin-resistant human head lice, Pediculus humanus capitis

Abstract.  The improper use of pediculicides containing permethrin has led to the development of resistance. Thus, new alternatives for control are needed. Plant-derived insecticides are attractive alternatives to common chemical insecticides because most of them are environmentally friendly and non-toxic to mammals. The toxic activity of 23 monoterpenoids belonging to several chemical classes was tested against the eggs of permethrin-resistant head lice, Pediculus humanus capitis De Geer (Phthiraptera: Pediculidae). Significant differences in ovicidal action were observed among the tested substances. The most effective chemicals were hydrocarbons and ethers, followed by ketones, alcohols, phenols and esters. A linear relationship between egg mortality and knockdown time (KT₅₀) on adults by the tested components revealed that most of the components were effective on both egg and adult stages. The monoterpenoids described herein are good candidates as effective pediculicides.     

The unusual reproductive system of head and body lice (Pediculus humanus)

Insect reproduction is extremely variable, but the implications of alternative genetic systems are often overlooked in studies on the evolution of insecticide resistance. Both ecotypes of Pediculus humanus (Phthiraptera: Pediculidae), the human head and body lice, are human ectoparasites, the control of which is challenged by the recent spread of resistance alleles. The present study conclusively establishes for the first time that both head and body lice reproduce through paternal genome elimination (PGE), an unusual genetic system in which males transmit only their maternally derived chromosomes. Here, we investigate inheritance patterns of parental genomes using a genotyping approach across families of both ecotypes and show that heterozygous males exclusively or preferentially pass on one allele only, whereas females transmit both in a Mendelian fashion. We do however observe occasional transmission of paternal chromosomes through males, representing the first known case of PGE in which whole‐genome meiotic drive is incomplete. Finally, we discuss the potential implications of this finding for the evolution of resistance and invite the development of new theoretical models of how this knowledge might contribute to increasing the success of pediculicide‐based management schemes.     

How long do louse eggs take to hatch? A possible answer to an age‐old riddle

There are no rigorous data on how long eggs of the head louse, Pediculus capitis (Phthiraptera: Pediculidae), take to hatch. Pediculicide users often report re‐infestations after apparently successful treatments in the absence of infective contacts. This study aimed to resolve the question of whether some louse eggs hatch after the completion of treatment, thereby giving rise to a new infestation. Data were extracted from the records of lice collected after treatments in 20 clinical intervention trials. All datasets were eliminated except those in which only newly hatched louse nymphs were found prior to the final assessment. This excluded the possibility that new eggs were laid after the first treatment and thus any young lice found must have originated from eggs laid before the start of treatment. This identified 23 of 1895 (1.2%) records with evidence of louse nymphs emerging at 13 days or more after the first treatment, 3–6 days longer than previous estimates. Current treatment regimens for pediculicides of two applications 7–10 days apart appear inadequate, which may explain continuing infestation in the community. Therefore, it is suggested that a revised approach using three treatments applied at intervals of 1 week should prevent the survival of any nymphs and their development into a new generation of adults.     

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.     

Permethrin-impregnated bednets reduce nuisance arthropods in Gambian houses

The prevalence of bedbugs (Cimex hemipterus L.), chicken ticks (Argas persicus Oken) and headlice (Pediculus capitis De Geer) was surveyed in a rural area of The Gambia. At the beginning of the study 37.5% of children's beds were infested with bedbugs and 3.9% with chicken ticks, whilst the prevalence rate of pediculosis in children under 10 years old was 28.8%. Both bedbugs and headlice were clustered within compounds. Headlice prevalence increased with hair length and they were more common on girls than boys. Following this cross-sectional survey all bednets in the sixteen hamlets were either dipped in permethrin or a placebo. About 4 months later it was found that bedbugs and chicken ticks had disappeared from homes in which the bednets had been impregnated with permethrin. There was no reduction in hamlets with placebo-treated bednets. The rate of acquiring headlice between the two surveys was reduced by 91.1% in children who slept under insecticide-treated bednets compared with children with placebo-treated bednets. There were also significantly fewer day-flying and crawling insects, except earwigs, in homes of children who slept under insecticide-treated bednets compared with those with placebo-treated nets. These additional benefits of permethrin-treated bednets should contribute to their widespread acceptance and utilization by the community for personal protection.     

Comparative trial of treatment with Prioderm lotion and Kwellada shampoo in children with head lice.

The safety and efficacy of treatment with a new pediculicide lotion, Prioderm (0.5% malathion in isopropanol), were compared with those of treatment with Kwellada (1% lindane) shampoo in a randomized trial of children with head lice. The children''s scalps were examined for live lice immediately, 7 days and 4 to 6 weeks following treatment. To determine the in-vitro ovicidal effect of treatment, samples of hair with nits were removed before and immediately following treatment; the subsequent rates of hatching were compared. No live lice were present immediately following either treatment. At 7 days after treatment 2 of the 29 children treated with Prioderm lotion and 4 of the 33 children treated with Kwellada shampoo were infested with live lice, whereas at 4 to 6 weeks after treatment 5 and 3 children respectively were infested. The initial effectiveness of treatment was 93% in the children treated with Prioderm lotion and 88% in those treated with Kwellada shampoo; however, a large difference in efficacy could have been missed owing to the small number of children in the study. Both preparations demonstrated in-vitro ovicidal activity, but neither totally abolished post-treatment hatching. No side effects were reported from either preparation. Because ovicidal activity was incomplete two treatments with the same pediculicides should be given about 7 days apart.     

Development of a paw paw herbal shampoo for the removal of head lice

The development and clinical testing of an herbal lice removal shampoo, containing a standardized extract of paw paw, thymol, and tea tree oil, are described. All of these ingredients were selected for their ability to deplete adenosine triphosphate (ATP) levels and, thus, prevent ATP-dependent pesticide resistance. Optimum concentrations of the ingredients, treatment times, and dosing schedules were established through in vitro tests with head lice. In addition to pilot studies involving 21 participants, a final clinical trial, using the optimum shampoo formulation in 16 participants, demonstrated 100% effectiveness in removing head lice and nits.     

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.     

A survey on head lice infestation in Korea (2001) and the therapeutic efficacy of oral trimethoprim/sulfamethoxazole adding to lindane shampoo

Total of 7,495 children including 3,908 boys and 3,587 girls from a kindergarten and 15 primary schools were examined for head lice infestation (HLI). The overall prevalence of HLI in this study was found to be 5.8%. Head lice were much more commonly detected in girls than in boys with prevalence of 11.2% and 0.9%, respectively. Sixty-nine children with HLI were treated with 1% lindane shampoo alone (group 1), and 45 children with HLI were treated with 1% lindane shampoo and oral trimethoprim/sulfamethoxazole (group 2), and follow-up visits were conducted 2 and 4 weeks later. The children who still had HLI 2 weeks after the primary treatment were treated again. At the 2-week follow-up visit, the treatment success rates of groups 1 and 2 were 76.8% and 86.7%, respectively, and at the 4-week follow-up visit, the rates were 91.3% and 97.8%, respectively. No statistically significant synergistic effect was observed for the combination of a 1% lindane shampoo and oral trimethoprim/sulfamethoxazole.     

Embryonic development of human lice: rearing conditions and susceptibility to spinosad

The embryonic development of human lice was evaluated according to the changes in the morphology of the embryo observed through the transparent chorion. Based on ocular and appendage development, three stages of embryogenesis were established: early, medium, and late. Influence of temperature and relative humidity (RH) on the laboratory rearing of Pediculus humanus capitis eggs was assessed. The optimal ranges for temperature and RH were 27-31 degrees C and 45-75%. The susceptibility of head [corrected] louse eggs to insecticide spinosad (a macrocyclic lactone) was assessed by immersion method. The results showed similar susceptibility to spinosad in early, medium, and late stages of head lice eggs. In addition, this study showed similar susceptibility of head and body lice eggs to spinosad, an insecticide that has not been used as pediculicide in Argentina (lethal concentration 50: 0.01%).     

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.