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.     

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.     

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 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.     

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.     

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.     

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.     

Characterization of body louse midgut proteins recognized by resistant hosts

Abstract. The human body louse, Pediculus humanus , showed eighteen midgut proteins ranging between 12 and 117 kDa, when analysed by SDS-PAGE electrophoresis. Seven of them (12 kDa, 17 kDa, 29 kDa, 35 kDa, 40 kDa, 55 kDa and 97 kDa) were major bands based on their intensity of staining. The immunization of rabbits with a midgut extract elicited the production of protective polyclonal antibodies. These antibodies reacted strongly with all major midgut proteins as well as with 63 kDa and 117 kDa proteins when tested by the Western blot technique. The analysis of the proteins revealed that the 12 kDa, 25 kDa, 29 kDa, 35 kDa, 45 kDa, 87 kDa and 97 kDa proteins are glycosylated and none of them contained a lipid moiety. By electroelution, the proteins of 35 kDa and 63 kDa were purified. On trypsinization, the proteins of 35 kDa and 63 kDa produced four major fragments (F, F₂, F₃, and F₄) when resolved on a 18% SDS-PAGE. The Fj fragment of the 35 kDa protein reacted with me polyclonal antibodies by the immunoblot technique.     

Repellency of essential oils against Nephotettix cincticeps: Laboratory and glasshouse assays

Four oils from Piper nigrum, Litsea cubeba, Zanthoxylum bungeanum and Curcuma longa were obtained by ethanol extraction. The repellency of these oils and two major compounds (linalool and piperine) was evaluated against female adult and third‐instar nymphs of the rice pest, Nephotettix cincticeps, under laboratory and glasshouse conditions. Paired‐choice and no‐choice assays were used for each treatment, with essential oils evaluated after 24 and 48 hr of exposure and chemical compounds evaluated after 12 and 24 hr of exposure. The potential effects of essential oils on activities of glutathione S tranferase (GST), carboxyl esterase (CarE) and acetyl cholinesterase (AchE) were also evaluated after 48 hr of exposure to leafhoppers. The constituents of the essential oils were determined using GC‐MS. The results showed that the major components in the oils were piperine (34.75%) for P. nigrum, 9,12‐octadecadienoic acid (Z,Z) (18.74%) for L. cubeba, ethanone, 1‐(2‐hydroxy‐4,6‐dimethoxyphenyl) (18.51%) for Z. bungeanum and turmerone (15.89%) for C. longa. In all cases, the essential oils repelled female adults and third‐instar nymphs of N. cincticeps. The repellency of the tested oils and chemicals compounds in the paired‐choice assay was higher than in the no‐choice assay. In all experimental conditions, P. nigrum and C. longa oils were the most and the least potent, respectively. Linalool was the best repellent among the single‐tested compounds under laboratory conditions. In the glasshouse study, the highest repellency was observed in the mixture of linalool and piperine. GST and CarE activities of leafhoppers were significantly enhanced by exposure to the four essentials oils; AchE activity increased significantly only in the P. nigrum and L. cubeba assays. Our results clearly indicate that the tested oils and chemical compounds are promising agents for developing plant‐based pesticides to control N. cincticeps.     

Chimeric mitochondrial minichromosomes of the human body louse, Pediculus humanus: evidence for homologous and non-homologous recombination

The mitochondrial (mt) genome of the human body louse, Pediculus humanus, consists of 18 minichromosomes. Each minichromosome is 3 to 4 kb long and has 1 to 3 genes. There is unequivocal evidence for recombination between different mt minichromosomes in P. humanus. It is not known, however, how these minichromosomes recombine. Here, we report the discovery of eight chimeric mt minichromosomes in P. humanus. We classify these chimeric mt minichromosomes into two groups: Group I and Group II. Group I chimeric minichromosomes contain parts of two different protein-coding genes that are from different minichromosomes. The two parts of protein-coding genes in each Group I chimeric minichromosome are joined at a microhomologous nucleotide sequence; microhomologous nucleotide sequences are hallmarks of non-homologous recombination. Group II chimeric minichromosomes contain all of the genes and the non-coding regions of two different minichromosomes. The conserved sequence blocks in the non-coding regions of Group II chimeric minichromosomes resemble the "recombination repeats" in the non-coding regions of the mt genomes of higher plants. These repeats are essential to homologous recombination in higher plants. Our analyses of the nucleotide sequences of chimeric mt minichromosomes indicate both homologous and non-homologous recombination between minichromosomes in the mitochondria of the human body louse.     

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.     

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.

---

     

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.     

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.     

Antifungal properties of essential oils and their main components uponCryptococcus neoformans

Cryptococcus neoformans opportunistic fungus met in the last phasis of AIDS is inhibited in vitro by several essential oils on natural volatile compounds. The minimal inhibitory concentration may reach 100 µl/l and minimal fungicidal concentration 200 µl/l with Palmarosa or Cinnamon oils. Among phenolic compounds, thymol and carvacrol are most fungitoxic. Terpenoids, citral, geraniol, and citronellol show best activities.     

The inhibition of Candida albicans by selected essential oils and their major components

Many volatile oils are known to possess antifungal properties and are potentially applicable as antimycotic agents. By studying the efficacy of essential oils against different pathogenic mycetes, we have evaluated the in-vitro inhibiting activity of some essential oils and their main constituents against a strain of Candida albicans. Sixteen commercial essential oils and forty-two pure constituents (alcohols, aldehydes, ketons, phenols and hydrocarbons), were tested by using a semisolid agar antifungal susceptibility (SAAS) method. Gas chromatography/mass spectroscopy analyses of the oils tested were performed. The essential oils of Origanum vulgare, Satureja montana, Mentha piperita, Cinnamomum verum, Cymbopogon flexuosus showed maximum inhibitory activity (MIC = 500 ppm) after 7 days. According to the results of the examination of pure constituents, -phellandrene proved to be the most interesting component among cyclic monoterpenic hydrocarbons as it showed a strong activity (MIC = 50 ppm). The most active of phenols was carvacrol (MIC 100 ppm). The open-chain alcohol 1-decanol was the most active of alcohols at 50 ppm. Finally, among aldehydes, a strong activity was shown by trans-cynnamaldehyde (MIC 50 ppm).     

Control of the chewing louse Bovicola (Werneckiella) ocellatus in donkeys,using essential oils

Infestations by lice can be a significant clinical and welfare issue in the management of large animals. The limited range of commercial pediculicides available and the development of resistance have led to the need to explore alternative louse management approaches. The results of in vitro and in vivo trials undertaken to control populations of the donkey chewing louse, Bovicola ocellatus (Piaget) (Phthiraptera: Trichodectidae) using the essential oils of tea tree (Melaleuca alternifolia) and lavender (Lavandula angustifolia) are reported here. Results of contact and vapour bioassays showed that 5% (v/v) tea tree and lavender oils resulted in > 80% louse mortality after 2 h of exposure. On farms, separate groups of 10 donkeys sprayed with 5% (v/v) tea tree and lavender oil as part of their usual grooming regime showed significant reductions in louse numbers compared with a control group (0.2% polysorbate 80 in water). These findings indicate that tea tree and lavender essential oils can provide clinically useful levels of control of B. ocellatus when used as part of a grooming routine and suggest that with further development could form the basis of an easy to apply and valuable component of a louse management programme for donkeys.