Measuring local genetic variation in permethrin-resistant head lice (Phthiraptera: Pediculidae) from Buenos Aires, Argentina.

The cosmopolitan ectoparasite human head louse, Pediculus humanus capitis (De Geer)(Phthiraptera:Pediculidae), affects mostly school-aged children, with infestations reported every year mainly due to louse resistance to pyrethroids. One of the main resistance mechanisms of pyrethroids is the target site insensitivity (kdr), which is caused by single-nucleotide point mutations (SNPs) located in the voltage-sensitive sodium channel gene. In this study, we analyzed individual head lice toxicologically via the description of their susceptibility profile to permethrin and genetically through the genotypification of their kdr alleles as well as nuclear microsatellite loci. Lice were collected from 4 schools in the city of Buenos Aires, Argentina. The resistance ratios varied from 33.3% to 71.4%, with a frequency of the T917I kdr mutation of 87.31% and with 83.6% of the head lice being homozygous resistant to pyrethroids. Microsatellite data indicated that all the louse school populations had genotype proportions that deviated from Hardy-Weinberg expectations, with FIS > 0 reflecting a deficit of heterozygotes. Bottleneck analysis suggested that all louse school populations underwent a recent reduction in population sizes, while 3 of the 4 schools had gene flow values around 1, indicating ongoing gene flow among those schools. Our study suggests that school louse populations in the city of Buenos Aires may form a metapopulation, where each school represents a small population that undergoes extinction and recolonization processes under strong permethrin selection. This is the first multilevel analysis integrating toxicological, kdr-genotyping, and microsatellite data in human louse populations.

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.

First exploratory study of bacterial pathogens and endosymbionts in head lice from a Mayan community in southern Mexico.

Lice represent one of the most neglected group of vectors worldwide, particularly in Latin America. Records of bacterial agents related to head lice are non-existent in this region of the continent. Many of these communities often do not have adequate access to public services and/or health protection. The normalization of this condition prevents them from manifesting discomfort, such as bites and itching, which further aggravates the situation, as they can be vectors of important diseases. For this reason, the aim of this work was to identify the richness of bacterial pathogens (Acinetobacter, Bartonella, and Rickettsia) and endosymbionts (Wolbachia) in head lice of paediatric patients from the indigenous municipality of Hoctun, Yucatan, Mexico. DNA extraction was performed using the QIAamp DNA Mini Kit. For the detection of bacterial pathogens, fragments of the gltA, rpoB, and 16S rDNA genes were amplified. For the detection of Wolbachia, the wsp gene was amplified. Of the 28 lice analysed, the presence of two genera of bacterial pathogens was detected Acinetobacter (42.9% = 12/28) and Bartonella (7.14% = 2/28). We also detected the endosymbiont Wolbachia (71.42% = 20/28). Our results showed that DNA from three bacteria species (Acinetobacter baumannii, Bartonella quintana, and Wolbachia pipientis) was present with frequencies ranging from 3.57% to 71.42%. This work represents the first exploratory study of the diversity of agents associated with head lice (Pediculus humanus capitis) in Mexico and Latin America. Due to the findings generated in the present study, it is important to perform surveillance of head lice populations to identify the degree of spread of these pathogens and their impact on populations in the region.

First evidence of the mutations associated with pyrethroid resistance in head lice (Phthiraptera: Pediculidae) from Honduras.

BACKGROUND: The human head louse, Pediculus humanus capitis, is a cosmopolitan blood-sucking ectoparasite affecting mostly schoolchildren in both developed and developing countries. In Honduras, chemical pediculicides are the first line of treatment, with permethrin as their main active ingredient. Despite the extended use of these products, there is currently no research investigating insecticide resistance in Honduran head lice. In head lice, the most common mechanism is knockdown resistance (kdr), which is the result of two point mutations and the associated amino acid substitutions, T917I and L920F, within the voltage-sensitive sodium channel (VSSC). METHODS: Genomic DNA was extracted from 83 head lice collected in the localities of San Buenaventura and La Hicaca, Honduras. Polymerase chain reaction (PCR) was used to amplify a 332-bp fragment of the VSSC gene that contains a site affected by C/T mutation which results in a T917I amino acid substitution on each human head louse genomic DNA fragments. RESULTS: The C/T non-synonymous mutation which results in the T917I kdr amino acid substitution was detected in both head lice populations at frequencies ranging between 0.45-0.5. Globally, the frequency of this substitution was 0.47. Of these, 5 (6.1%) were homozygous susceptible and 78 (93.9%) were heterozygotes. The kdr-resistant homozygote (RR) was not detected in the studied populations. Thus, 93.9% of the head lice collected in Honduras harbored only one T917I allele. Exact test for the Hardy-Weinberg equilibrium for both localities showed that genotype frequencies differed significantly from expectation. In addition, San Buenaventura and La Hicaca populations had an inbreeding coefficient (Fis) < 0, suggesting an excess of heterozygotes. CONCLUSIONS: To our knowledge, this is the first study showing the presence of the C/T mutation responsible of the T917I kdr allele associated with pyrethroid resistance in P. h. capitis from Honduras. The PCR-restriction fragment length polymorphism (RFLP) employed here has demonstrated to be a reliable, economic, and reproducible assay that can be used to accurately genotype individual head lice for the mutation encoding the resistance-conferring T917I amino acid substitution. This highlights the necessity of proactive resistance management programmes designed to detect pyrethroid mutations before they become established within populations of head lice.

First Determination of Pyrethroid Knockdown Resistance Alleles in Human Head Lice (Phthiraptera: Pediculidae) From Chile.

The infestation with the human ectoparasite, Pediculus humanus capitis (De Geer), is a common public health problem affecting schoolchildren worldwide. In Chile, the main active ingredients present in the over-the-counter pediculicides contain pyrethroids. Despite the extended use of these products, there is no evidence of the insecticide resistance status of the head lice geographically located in Chile. The most extended resistant mechanism of pyrethroids consists of the target site insensitivity (Kdr) determined by the presence of mutations linked to insecticide-binding sites in the voltage-sensitive sodium channel. T917I is recognized as the main mutation in head lice, and detection is considered to be a biomarker of resistance. The goal of the present study was to detect the presence and distribution of T917I mutation in five geographic locations of Chile. All five geographically selected louse populations had a frequency of pyrethroid resistance genes that ranged from 36 to 77%, and 94.9% of the collected head lice had one or two T917I mutant alleles. Moreover, the frequency of the aggregate resistant alleles was 50.5%. This is the first evidence that head lice in Chile had the mutations commonly associated with the resistance to pyrethroids. Moreover, the overrepresentation of heterozygotes in the studied populations suggests that head lice in Chile are currently under active selective pressure.

Scalp microbiota alterations in children with pediculosis.

Pediculosis is a disease caused by the insect Pediculus humanus capitis that mainly occurs in childhood. A comparative study was carried out evaluating groups of schoolchildren with (group A) and without pediculosis (group B) to analyse the characteristics of the scalp microbiota. Samples were collected by swab using Stuart transport medium and incubate in Sabouraud dextrose agar with tetracycline to analyse the fungal microbiota and in blood agar to assess the bacterial microbiota. The isolates identity was confirmed by sequencing of the 16S and 18S regions of the ribosomal DNA gene for bacteria and fungi, respectively. The analysis of the 186 isolates led to the identification of 35 bacteria and 40 fungi in group A and 47 bacteria and 64 fungi in group B. The results indicate differences in bacterial and fungal species in the groups analysed. In the observed bacterial microbiota, Staphylococcus capitis occurred more frequently than Staphylococcus epidermidis in group A vs B. Among fungal isolates, Debaryomyces sp. was more frequent in group B vs A. Our findings showed scalp microbiota alterations in children with pediculosis, meriting future studies to analyse the relationship between these agents and their impact on human health.

Mitochondrial diversity and phylogeographic analysis of Pediculus humanus reveals a new Amazonian clade "F".

Pediculus humanus is an obligate and highly intimate bloodsucking insect parasite of humans that has two ecotypes, head louse and body louse. This study analyzed genetic diversity at three mitochondrial genes (cytochrome b [cytb], cytochrome oxidase subunit 1 [cox1] and 12S ribosomal RNA [12S]) in 98 head lice collected from an isolated Native American population from the Wayampi community in Trois-Sauts, French Guiana. These results are integrated with all prior data of P. humanus (1402 cytb, 743 cox1 and 344 12S) from other parts of the world. The phylogenetic analysis revealed six highly divergent and well-supported monophyletic clades. Five clades corresponded to the previously recognized mitochondrial clades A, D, B, C and E, while the sixth (clade F) was novel, as it exhibited 5.4%, 3.7% and 3.6% divergence at cytb, cox1 and 12S, respectively, from its nearest neighbor clade B. Interestingly, the clade F has only been recovered in a few lice sequences from Mexico and Argentina, while it was the most common lineage in the Amazonian lice, which hints its association with the Native American region. Furthermore, Pediculus mjobergi, a New World monkeys' louse, which is thought to be transmitted to monkeys from the first humans that had reached the American continent thousands of years ago, also belonged to this clade, suggesting that this louse may not be a separate species but an evolutionary lineage of P. humanus. The discovery of new Amazonian clade F with the recovery of additional haplotypes within each of the five clades demonstrates that the levels of genetic diversity in P. humanus are higher than previously thought.

First Detection of the Kdr Mutation T929I in Head Lice (Phthiraptera: Pediculidae) in Schoolchildren of the Metropolitan Area of Nuevo Leon and Yucatan, Mexico.

The head louse Pediculus humanus capitis (De Geer) is a hematophagous ectoparasite that inhabits the human scalp. Infestations by this insect are commonly known as pediculosis, which is more common in younger groups. These infestations are asymptomatic; however, skin irritation from scratching occasionally may cause secondary bacterial infections. In recent years, the prevalence of pediculosis has increased in children; this increase has been attributed to louse resistance to the insecticides used as a control measure for infestation. The aim of the present study was to determine the presence and frequency of the knockdown resistance mutation (kdr) T929I in 468 head lice collected from 32 elementary schools in the metropolitan area of Nuevo Leon (24) and Yucatan (8), Mexico. This is the first report of a knockdown resistance (kdr) mechanism in head lice from Mexico. The T929I mutation was present in all of the sampled schools, with variability observed in its allelic and genotypic frequencies.

Cytogenetic Features of Human Head and Body Lice (Phthiraptera: Pediculidae).

The genus Pediculus L. that parasitize humans comprise two subspecies: the head lice Pediculus humanus capitis De Geer and the body lice Pediculus humanus humanus De Geer. Despite the 200 yr of the first description of these two species, there is still a long debate about their taxonomic status. Some authors proposed that these organisms are separate species, conspecifics, or grouped in clades. The sequencing of both forms indicated that the difference between them is one gene absent in the head louse. However, their chromosomal number remains to be determined. In this study, we described the male and female karyotypes, and male meiosis of head and body lice, and examined the chromatin structure by means of C-banding. In P. h. humanus and P. h. capitis, the diploid chromosome complement was 2 n = 12 in both sexes. In oogonial prometaphase and metaphase and spermatogonial metaphase, it is evident that chromosomes lack of a primary constriction. No identifiable sex chromosomes or B chromosomes were observed in head and body lice. Neither chiasmata nor chromatin connections between homologous chromosomes were detected in male meiosis. The meiotic behaviour of the chromosomes showed that they are holokinetic. C-banding revealed the absence of constitutive heterochromatin. Our results provide relevant information to be used in mapping studies of genes associated with sex determination and environmental sensing and response.

Geographical distribution of pyrethroid resistance allele frequency in head lice (Phthiraptera: Pediculidae) from Argentina.

The human head louse, Pediculus humanus capitis De Geer (Phthiraptera: Pediculidae), is an obligate ectoparasite that causes pediculosis capitis and has parasitized humans since the beginning of humankind. Head louse infestations are widespread throughout the world and have been increasing since the early 1990s partially because of ineffective pediculicides. In Argentina, the overuse of products containing pyrethroids has led to the development of resistant louse populations. Pyrethroid insecticides act on the nervous system affecting voltage-sensitive sodium channels. Three point mutations at the corresponding amino acid sequence positions M815I, T917I, and L920F in the voltage-gated sodium channel gene are responsible for contributing to knockdown resistance (kdr). The management of pyrethroid resistance requires either early detection or the characterization of the mechanisms involved in head louse populations. In the current study, we estimated the distribution of kdr alleles in 154 head lice from six geographical regions of Argentina. Pyrethroid resistance kdr alleles were found in high frequencies ranging from 67 to 100%. Of these, 131 (85.1%) were homozygous resistant, 13 (8.4%) were homozygous susceptible, and 10 (6.5%) were heterozygous. Exact tests for the Hardy-Weinberg equilibrium for each location showed that genotype frequencies differed significantly from expectation in four of the six sites studied. These results show that pyrethroid resistance is well established reaching an overall frequency of 88%, thus close to fixation. With 30 yr of pyrethroid-based pediculicides use in Argentina, kdr resistance has evolved rapidly among these head louse populations.

Evidence of sympatry of clade a and clade B head lice in a pre-Columbian Chilean mummy from Camarones.

Three different lineages of head lice are known to parasitize humans. Clade A, which is currently worldwide in distribution, was previously demonstrated to be present in the Americas before the time of Columbus. The two other types of head lice are geographically restricted to America and Australia for clade B and to Africa and Asia for clade C. In this study, we tested two operculated nits from a 4,000-year-old Chilean mummy of Camarones for the presence of the partial Cytb mitochondrial gene (270 bp). Our finding shows that clade B head lice were present in America before the arrival of the European colonists.

Amazonian head lice-specific genotypes are putatively pre-Columbian.

Head and body lice are strict obligate human ectoparasites with three mitochondrial phylotypes (A, B, and C). Using molecular methods for genotyping lice (Cytochrome b and multi-spacer typing), and comparing our results with all the sequences of human lice that were genotyped previously, we assessed the presence of a specific American genotype that most likely predates the Columbian era in head lice collected from Amazonia.

Molecular identification of lice from pre-Columbian mummies.

BACKGROUND: Three distinctly different lineages of head and body lice are known to parasitize humans. One lineage includes head and body lice and is currently worldwide in distribution (type A). The other 2 (types B and C) include only head lice and are geographically restricted. It was hypothesized that head louse phylotypes were exchanged only recently, after European exploration and colonization (after Columbus). METHODS: To determine which louse type or types were found in the Americas before European colonization, we used polymerase chain reaction in 2 laboratories to amplify DNA from 2 genes (Cytb and Cox1) belonging to 1000-year-old lice collected from Peruvian mummies. RESULTS: Only the worldwide type (type A) was found. Therefore, this phylotype was worldwide before European colonization, as type A lice were common in Europe, Africa, and Asia. CONCLUSIONS: The findings of this study show that several phylotypes of head lice have coexisted for centuries in humans and support the claim that type A lice were present in the Americas before the time of Columbus.