Interspecific hybridization of Eucalyptus as a potential tool to improve the bioactivity of essential oils against permethrin-resistant head lice from Argentina

The essential oils extracted from Eucalyptus grandis, Eucalyptus camaldulensis, Eucalyptus tereticornis, and the hybrids E. grandisxE. camaldulensis, and E. grandisxE. tereticornis were analyzed by GC-MS, and evaluated for their fumigant and repellent effects on permethrin-resistant head lice. Fumigant activity of both hybrids was higher than that for pure species. E. grandisxE. tereticornis and E. grandisxE. camaldulensis showed KT50 values of 12.99 and 13.63min, respectively. E. grandis, E. camaldulensis, and E. tereticornis showed KT50 values of 25.57, 35.01, and 31.31, respectively. A simple regression analysis revealed a significant correlation between KT50 data and % of 1,8-cineole in these essential oils. Repellency varied from 47.80+/-16% to 80.69+/-6% for the five Eucalyptus essential oils tested. Interspecific hybridization improves the pediculicidal activity of Eucalyptus essential oils.     

Insecticidal activity of essential oils from eleven Eucalyptus spp. and two hybrids: lethal and sublethal effects of their major components on Blattella germanica

The aim of the current study was to evaluate the fumigant activity of the essential oils from 11 species of the genus Eucalyptus and two of their hybrids on first instar of Blattella germanica L. The fumigant activity and repellence of the four major monoterpene components of these essential oils also were tested. Fumigant activity was evaluated by exposing nymphs to the vapors emitted by 50 microl of essential oil or monoterpene in a closed container. The lowest knockdown time 50% (KT50) values, expressed in minutes, were elicited by the essential oils of the Eucalyptus grandis X Eucalyptus tereticornis (57.9) hybrid, Eucalyptus sideroxylon A. Cunn (62.0), E. grandis X Eucalyptus camaldulensis (63.8) hybrid, Eucalyptus viminalis Labill (64.1), Eucalyptus dunnii Maiden (64.5), and Eucalyptus grandis (Hill) ex Maiden (68.7). The KT50 values for the remaining essential oils ranged between 74.5 (E. saligna Smith) and 161.4 min (E. tereticornis Smith). The essential oil from the hybrid E. grandis X E. tereticornis was 3.7 times less toxic than dichlorvos (positive control). The KT50 values of monoterpenes were 38.8 for alpha-pinene, 55.3 for 1,8-cineole, 175.6 for p-cymene, and 178.3 for gamma-terpinene. Alpha-pinene was 2.5 times less toxic than dichlorvos. There was a strong positive correlation between the fumigant activity of essential oils and their corresponding 1,8-cineole and alpha-pinene concentration. Repellency was quantified using a video tracking system. Two concentrations of monoterpenes were studied (7 and 70 microg/cm2). All compounds produced a light repellent effect but only when applied at 70 microg/cm2. In all cases, the repellent effect was less than that produced by the broad-spectrum insect repellent N,N-diethyl-3-methylbenzamide (positive control).     

Evidence That Head and Body Lice on Homeless Persons Have the Same Genotype

Human head lice and body lice are morphologically and biologically similar but have distinct ecologies. They were shown to have almost the same basic genetic content (one gene is absent in head lice), but differentially express certain genes, presumably responsible for the vector competence. They are now believed to be ecotypes of the same species (Pediculus humanus) and based on mitochondrial studies, body lice have been included with head lice in one of three clades of human head lice (Clade A). Here, we tested whether head and body lice collected from the same host belong to the same population by examining highly polymorphic intergenic spacers. This study was performed on lice collected from five homeless persons living in the same shelter in which Clade A lice are prevalent. Lice were individually genotyped at four spacer loci. The genetic identity and diversity of lice from head and body populations were compared for each homeless person. Population genetic structure was tested between lice from the two body regions and between the lice from different host individuals.We found two pairs of head and body lice on the same homeless person with identical multi locus genotypes. No difference in genetic diversity was found between head and body louse populations and no evidence of significant structure between the louse populations was found, even after controlling for a possible effect of the host individual. More surprisingly, no structure was obvious between lice of different homeless persons.We believe that the head and body lice collected from our five subjects belong to the same population and are shared between people living in the same shelter. These findings confirm that head and body lice are two ecotypes of the same species and show the importance of implementing measures to prevent lice transmission between homeless people in shelters.     

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%).     

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.     

Prevalence of head lice (Pediculus humanus capitis) infestation among schools workers in the Eastern Region,Saudi Arabia

In a cross-sectional survey in February-May 2019, the prevalence of Pediculosis capitis with demographic data and the behavioral practices were investigated among 750 participants in the Eastern region of Saudi Arabia. Female participation was highly remarked with a percentage of 94.08% compared to that of male one about 5.91%. A deficiency of knowledge about lice infestation was noted especially among illiterate participants raised from their socio-economic levels (p-value = 0.001). Lice infestation reached higher rates in children aged less than 20 years with itching of the hair scalp. The obtained results revealed that 59.33% of the respondents believed that frequency of personal hygiene and washing of hair were the best methods for preventing the lice infestation. However, the treatment of lice infestation using anti-lice agents (p-value = 0.020) was preferred by 14.26% of participants. Although knowledge about the preventive tools for lice infestation (p-value 0.089) was not significantly associated with the experience of infestation but knowledge about the appropriate treatment to kill lice (p-value 0.020) and the wrong practices in the treatment of a head lice infestation were (p-value 0.005) significantly associated with the experience of infestation. Health programs and campaigns preventions are highly advised to increase the awareness of Pediculosis capitis with an effective strategic plan to control, manage, and prevent this disease.     

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.     

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.     

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.     

Fumigant Toxicity of Plant Essential Oils to Plutella xylostella (Lepidoptera: Yponomeutidae) and Cotesia glomerata (Hymenoptera: Braconidae)

The fumigant toxicity of 66 plant essential oils to Plutella xylostella (L.) larvae and Cotesia glomerata (L.) adults was examined using a vapor-phase toxicity bioassay and compared with that of dichlorvos. Responses varied according to oil and insect species used. Based on 24 h LD₅₀ values, pennyroyal oil [10.77 mg/filter paper (4.25 cm diameter)] was the most toxic fumigant, followed by rosemary and sage (Dalmatin) oils (15.15 mg/paper). Potent fumigant toxicity was also produced from armoise, buchu leaf, cedarleaf, coriander, eucalyptus, howood, lavender, myrtle, niaouli, peppermint, and rosewood oils (LD₅₀, 21.29–27.31 mg/paper). All essential oils were less effective than dichlorvos (LD₅₀, 0.52 mg/paper). Against adult C. glomerata, dichlorvos (LD₅₀, 0.03 mg/paper) was the most toxic fumigant, whereas the LD₅₀ values of the 14 essential oils ranged from 1.59 to 8.51 mg/paper. Based on selective toxicity ratio (STR, P. xylostella LD₅₀/C. glomerata LD₅₀), the 14 essential oils (STR, 2.5–14.5) are more selective than dichlorvos (STR, 17.3). The essential oils tested merit further study as potential fumigants for the control of P. xylostella in greenhouses because of their selective toxicity to adult C. glomerata and their much greater activity as a fumigant.     

双去甲氧基姜黄素对朱砂叶螨的生物活性及作用方式

在室温26℃±1℃、湿度60%～80%、光照14 h条件下,测定了天然植物活性成分双去甲氧基姜黄素对重要植食性害螨朱砂叶螨各螨态的触杀和熏蒸活性,对幼螨、若螨、成螨的驱避活性,以及对雌成螨的产卵抑制活性.采用玻片浸渍法测得双去甲氧基姜黄素对朱砂叶螨雌成螨48 h的LCso为0.433 mg·mL-1.在0.883 mg·mL-1(LC70)的浓度下,双去甲氧基姜黄素对朱砂叶螨不同螨态触杀活性的大小依次为幼螨＞若螨＞成螨＞卵,其中对幼螨24h和48 h的校正死亡率分别为60.0%和83.3%;对朱砂叶螨各螨态的熏蒸作用不明显,024h和48 h的校正死亡率均小于3%;对幼螨、若螨以及成螨均表现出较强的驱避作用,其中对幼螨的效果最好,不同处理时间的驱避率均在85%以上,其次是若螨,对成螨的驱避性相对较差,72 h的驱避率仅为47.8%.同时对雌成螨有明显的产卵抑制作用,处理后120 h产卵抑制率达到89.3%.表明双去甲氧基姜黄素对朱砂叶螨的主要作用方式为触杀、驱避和产卵抑制作用.     

D-limonene nanoemulsion: lousicidal activity,stability, and effect on the cuticle of Columbicola columbae

The current study was conducted to investigate the efficacy and stability of D-limonene (DL) and its nanoemulsion (DLN) against pigeon feather lice (Columbicola columbae) and their mode of action. DL pure form and DLN were prepared and characterized freshly and after storage for 50 days. In vitro bioassay on live lice was conducted with different concentrations of DL, DLN, and deltamethrin (DM). The results revealed significant mortality rates in the DL-, DLN-, DM-treated groups when compared with the control (p < 0.05). The scanning electron micrographs of lice treated with DL and DLN revealed collapsed bodies with destruction in the cuticle of the mouthparts and damaged antennae. The 50 days stored DLN showed stability in their effectiveness when compared with the freshly prepared formulation. DL and DLN caused significant inhibition (p ≤ 0.05) in acetylcholinesterase activity (AchE). Malondialdehyde level (MDA) was significantly increased while glutathione was significantly decreased in DL- and DLN-treated lice. In conclusion, DL and DLN have significant lousicidal activities. DLN showed better stability than DL after storage for 50 days. In addition, the mode of action of DL may associate with its effect on the cuticle of the lice body, inhibition of AchE, and increasing oxidative stress in the treated lice.     

Resistance and the control of lice on humans and production animals

Phthiraptera (lice) are specialised insects adapted to parasitise many warm-blooded vertebrates, including domestic animals and humans. Often, attempts by the host to alleviate the irritation created by lice, causes derangement of the hair/fur coat. Unless treated, this derangement may cause economic losses due to hide damage and/or downgrading of wool/hair/fur. In 1981, application of aqueous insecticide solutions (dipping) for the control of sheep body lice (Bovicola ovis) was largely superseded by off-shears pyrethroid "pour-on" treatments. By 1985, several field failures with these products were found to be due to low-level (20x) insecticide resistance. In 1990, high-level (640x) resistance was diagnosed in a New South Wales population. However, despite 30+years use, organophosphate-based products are still usually effective. Until recently, cattle lice caused little concern. Treatments were applied mainly for aesthetic reasons when cattle were to be presented for sale, and also to prevent damage to fences by rubbing cattle. However, the introduction of quality-management programmes have raised awareness of the economic losses due to hide damage associated with lice infestations. Emerging industries such as emu and alpaca farming have raised the pest status of other louse species, and necessitated insecticidal intervention. In humans, attempts to control head lice, Pediculus humanus capitis, infestations have repeatedly failed around the world.     

Ovicidal and adulticidal activities of Cinnamomum zeylanicum bark essential oil compounds and related compounds against Pediculus humanus capitis (Anoplura: Pediculicidae)

The toxicity of cinnamon, Cinnamomum zeylanicum, bark essential oil compounds against eggs and adult females of human head louse, Pediculus humanus capitis, was examined using direct contact and vapour phase toxicity bioassays and compared with the lethal activity of their related compounds, benzyl alcohol, cinnamic acid, cinnamyl acetate, 4-hydroxybenzaldehyde and salicylaldehyde, as well as two widely used pediculicides, d-phenothrin and pyrethrum. In a filter-paper contact toxicity bioassay with female lice at 0.25 mg/cm², benzaldehyde was 29- and 27-fold more toxic than pyrethrum and d-phenothrin, respectively, as judged by median lethal time (LT₅₀) values. Salicylaldehyde was nine and eight times more active than pyrethrum and d-phenothrin, respectively. Pediculicidal activity of linalool was comparable with that of d-phenothrin and pyrethrum. Cinnamomum bark essential oil was slightly less effective than either d-phenothrin or pyrethrum. Benzyl alcohol and (E)-cinnamaldehyde exhibited moderate pediculicidal activity. After 24 h of exposure, no hatching was observed with 0.063 mg/cm² salicylaldehyde, 0.125 mg/cm² benzaldehyde, 0.5 mg/cm² Cinnamomum bark essential oil, 1.0 mg/cm² (E)-cinnamaldehyde, and 1.0 mg/cm² benzyl cinnamate. Little or no ovicidal activity was observed with d-phenothrin or pyrethrum. In vapour phase toxicity tests with female lice, benzaldehyde and salicylaldehyde were much more effective in closed containers than in open ones, indicating that the mode of delivery of these compounds was largely due to action in the vapour phase. Neither d-phenothrin nor pyrethrum exhibited fumigant toxicity. Cinnamomum bark essential oil and test compounds described merit further study as potential pediculicides or ovicides for the control of P. h. capitis.     

The geographical segregation of human lice preceded that of Pediculus humanus capitis and Pediculus humanus humanus

In order to investigate human-louse phylogeny, we partially sequenced two nuclear (18S rRNA and EF-1 alpha) and one mitochondrial (COI) genes from 155 Pediculus from different geographical origins. The phylogenetic analysis of 18S rRNA and EF-1 alpha sequences showed that human lice were classified into lice from Sub-Saharan Africa and lice from other areas. In both clusters, head and body lice were clearly grouped into two separate clusters. Our results indicate that the earliest divergence within human pediculidae occurred between African lice and other lice, and the divergence between head and body lice was not the result from a single event.     

石菖蒲根茎甲醇提取物对谷蠹的生物活性

采用冷浸法和溶剂萃取法,研究石菖蒲(Acorus gramineus Soland)根茎甲醇提取物对谷蠹Rhizopertha dominica(Fabricius)成虫的生物活性。结果表明,石菖蒲甲醇提取物以石油醚萃取物的生物活性最强,水层次之。石油醚萃取物对谷蠹具有较高的触杀、熏蒸、种群抑制活性和防治效果,并具有一定的驱避活性。药膜法触杀处理72h后对谷蠹的LC50和LC95分别为8.30μg/cm2,51.75μg/cm2。药纸熏蒸法6.25μL/L处理120h后对谷蠹的校正死亡率为94.25%。饲料拌药法250mg/kg处理对谷蠹的防效与5mg/kg马拉硫磷效果相当,且对F1代的种群抑制效果显著高于5mg/kg马拉硫磷处理。     

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 head and body lice of humans are genetically distinct (Insecta: Phthiraptera, Pediculidae): evidence from double infestations

Little is known about the population genetics of the louse infestations of humans. We used microsatellite DNA to study 11 double infestations, that is, hosts infested with head lice and body lice simultaneously. We tested for population structure on a host, and for population structure among seven hosts that shared sleeping quarters. We also sought evidence of migration among louse populations. Our results showed that: (i) the head and body lice on these individual hosts were two genetically distinct populations; (ii) each host had their own populations of head and body lice that were genetically distinct to those on other hosts; and (iii) lice had migrated from head to head, and from body to body, but not between heads and bodies. Our results indicate that head and body lice are separate species.     

Evaluation of three methods for detecting permethrin resistance in adult and nymphal Blattella germanica (Dictyoptera: Blattellidae).

The insecticide resistance status of the German cockroach, Blattella germanica (L.), against permethrin was evaluated with three different bioassay methods. The LT50 and KT50 in adults were measured by glass jar test, the LT50 and KT50 in nymphs were assessed by glass petri dish-test, and the LD50 in adults and nymphs was measured by topical application. The results indicate that in all three bioassays, cockroaches showed different degrees of resistance to permethrin. Resistance ratios of 6.7-, 12.8-, and 7.5-fold for KT50, LD50, and LT50, respectively, were obtained for adult cockroaches compared with those from a susceptible strain. Similarly, resistance ratio values of 17-fold for KT50, 8-fold for LD50, and 6.4-fold for LT50 were obtained for nymphs compared with those from a susceptible strain. Although both adults and nymphs exhibited a similar level of susceptibility to permethrin, adults showed a higher degree of susceptibility to topical application than to the other exposure method. The order of sensitivity (based on resistance ratio values) was topical application (LD50) > LT50 > KT50 in adults. For nymphs, the resistance ratio in the knockdown test was higher than in other tests (resistance ratio = 17-fold). Therefore, the order of test sensitivity based on comparisons of resistance ratio values was KT50 > topical application (LD50) > LT50 in nymphs. Topical application exhibited a higher sensitivity than the LT and KT methods in adults. Thus, the results suggest that topical application and knockdown testing methods are the most sensitive assays for measuring resistance of adult and nymphal German cockroaches, respectively, to permethrin.     

Distinguishing Body Lice from Head Lice by Multiplex Real-Time PCR Analysis of the Phum_PHUM540560 Gene

Background

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.

Materials and Methods

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.

Results

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.

Conclusions

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