Comparative Effects of Elettaria cardamomum Essential Oil and Its Nanoliposomal State on Mortality of Anopheles stephensi Larvae.

Background: Malaria has remained the most dreadful vector-borne disease; hence, vector control is the most affordable and achievable approach to mitigate the disease burden. Due to the emergence of resistance and environmental pollution, herbal larvicides are considered an alternative to chemical types. Also, nanotechnology has been proposed as a promising solution to improve the efficiency of plant larvicides. This study aimed to develop an effective herbal larvicide. Methods: The chemical composition of Elettaria cardamomum essential oil (EO) was first investigated. Nanoliposomes containing the EO were then prepared using the ethanol injection method. After that, the larvicidal efficacy of the EO and its liposomal state were compared against Anopheles stephensi in laboratory conditions. Results: Alpha-terpinyl acetate (77.59%), eucalyptol (4.38%), nerolidol (2.96%), linalool (1.77%), and limonene (1.69%) were the five major compounds of the EO. Nanoliposomes containing the EO with a particle size of 73±5 nm and a zeta potential of -16.3±0.8 mV were prepared. Additionally, the ATR-FTIR analysis verified the successful loading of the EO into nanoliposomes. The larvicidal activity of nanoliposomes exhibited remarkable potency, with an LC50 value of 14.35 (10-18) µg/mL, significantly more potent than the non-formulated EO, which had an LC50 value of 33.47 (28-39) µg/mL against Anopheles stephensi larvae. Conclusion: The nanoliposomes containing E. cardamomum EO showed promising efficacy against An. stephensi larvae. It could thus be considered for further application against other species of mosquitoes.

Identification, molecular characterization, and in silico structural analysis of larval salivary glands Netrin-A as a potent biomarker from Lucilia sericata (Diptera: Calliphoridae).

The greenbottle blowfly Lucilia sericata (L. sericata) is increasingly used in larval therapy of chronic wounds. Netrins as bifunctional proteins are in the superfamily of Laminins secreted from larval salivary glands. The Netrin protein has a significant instructive role in axon guidance, causing neuronal outgrowth, angiogenesis, and cell migration. It seems to be crucial in wound healing and acts as a potential biomarker in diagnosing some clinical diseases. This survey aimed to identify molecular features and analyze in silico structural configuration of Netrin-A in L. sericata larvae. The larvae were reared under standard maggotarium conditions. The nucleic acid sequence of L. sericata Netrin-A (LSN-A) was then identified using rapid amplification of circular DNA ends (RACE) and rapid amplification of genomic ends (RAGE). Parts of the Netrin-A gene, including the middle, 3'-, and 5'-ends, were identified, TA cloned in pTG19 plasmid, and transferred into DH5ɑ Escherichia coli. Each part was sequenced and assembled using SeqMan software. This gene structure was further subjected to in silico analysis. The DNA of LSN-A was identified to be 2407 bp, while its mRNA sequence was recognized as 2115 bp by Oligo0.7 software. It translated the Netrin-A protein with 704 amino acid residues. Its estimated molecular weight was 78.6 kDa. Sequencing of this fragment and its BLAST analysis revealed laminin-based high (95%) similarity with the mRNA sequence of Lucilia cuprina Netrin-A. The 3-D structure of Netrin-A drawn by SWISS-MODEL exhibited its partial resemblance to the reference molecule Netrin-1 of Homo sapiens. This study supports the molecular and structural analyses of LSN-A protein, which could lead to wound treatment. Ultimately, it can be an effective candidate to ameliorate injury. Our next attempt is to produce LSN-A recombinant protein for use in biomedical sciences.

Frequency of pyrethroid resistance in human head louse treatment: systematic review and meta-analysis.

Head lice (Pediculus humanus capitis) are one of the most common insects causing infestations in humans worldwide, and infestation is associated with adverse socio-economic and public health effects. The development of genetic insensitivity (e.g., target site insensitivity = knockdown resistance or kdr) to topical insecticides has impaired effective treatment. Therefore, this study was undertaken to review and meta-analyze the frequency of pyrethroid resistance in treated head louse populations from the beginning of 2000 to the end of June 2021 worldwide. In order to accomplish this, all English language articles published over this period were extracted and reviewed. Statistical analyses of data were performed using fixed and random effect model tests in meta-analysis, Cochrane, meta-regression and I2 index. A total of 24 articles from an initial sample size of 5033 were accepted into this systematic review. The mean frequency of pyrethroid resistance was estimated to be 76.9%. In collected resistant lice, 64.4% were homozygote and 30.3% were heterozygote resistant. Globally, four countries (Australia, England, Israel, and Turkey) have 100% kdr gene frequencies, likely resulting in the ineffectiveness of pyrethrin- and pyrethroid-based pediculicides. The highest resistance recorded in these studies was against permethrin. This study shows that pyrethroid resistance is found at relatively high frequencies in many countries. As a result, treatment with current insecticides may not be effective and is likely the cause of increased levels of infestations. It is recommended that resistance status be evaluated prior to insecticide treatment, to increase efficacy.

TITLE: Fréquence de la résistance aux pyréthroïdes dans le traitement du pou de tête chez l'homme : revue systématique et méta-analyse. ABSTRACT: Les poux de tête (Pediculus humanus capitis) sont l'un des insectes les plus courants à l'origine d'infestations chez l'homme dans le monde, et l'infestation est associée à des effets socio-économiques et de santé publique néfastes. Le développement d'une insensibilité génétique (par exemple, l'insensibilité au site cible = résistance knockdown ou kdr) aux insecticides topiques a altéré l'efficacité de leur traitement. Par conséquent, cette étude a été entreprise pour examiner et méta-analyser la fréquence de la résistance aux pyréthroïdes dans les populations de poux de tête étudiées du début 2000 à la fin juin 2021 dans le monde. Pour ce faire, tous les articles en anglais publiés au cours de cette période ont été extraits et examinés. Les analyses statistiques des données ont été effectuées à l'aide de tests de modèles à effets fixes et aléatoires dans la méta-analyse, Cochrane, méta-régression et indice I2. Un total de 24 articles provenant d'un échantillon initial de 5033 ont été acceptés dans cette revue systématique. La fréquence moyenne de la résistance aux pyréthroïdes a été estimée à 76,9 %. Chez les poux résistants collectés, 64,4 % étaient homozygotes résistants et 30,3 % étaient hétérozygotes résistants. À l'échelle mondiale, quatre pays (Australie, Angleterre, Israël et Turquie) ont des fréquences de gène kdr de 100 %, ce qui entraîne probablement une inefficacité des pédiculicides à base de pyréthrine et de pyréthrinoïde. La résistance la plus élevée enregistrée dans ces études était celle contre la perméthrine. Cette étude montre que la résistance aux pyréthroïdes est trouvée à des fréquences relativement élevées dans de nombreux pays. En conséquence, le traitement avec les insecticides actuels peut ne pas être efficace et est probablement la cause d'une augmentation des niveaux d'infestation. Il est recommandé d'évaluer le statut de résistance avant le traitement insecticide, pour augmenter son efficacité.

Potential co-infection of Wolbachia with Leishmania among sand fly vectors caught from endemic leishmaniasis foci in Fars province, southern Iran.

Leishmaniasis is one of the Neglected Tropical Diseases in the tropical region of many countries in the world. The etiological agents (Leishmania parasites) of the disease are transmitted to human and other vertebrate hosts by infectious bites of female phlebotomine sand flies. On the other hand, some symbiotic microorganisms such as Wolbachia (Rickettsiales: Anaplasmataceae) may be transmitted vertically in many arthropods and may cause synergistic or antagonistic effects on epidemiology of the vector-borne diseases. Hence, in the present study, potential coinfection of Wolbachia with Leishmania in the sand fly vectors will be examined by PCR technique in the important leishmaniasis foci of Fars province in southern Iran, as a new feature for the disease long-term control. Sand flies were collected by sticky traps from indoor and outdoor locations of 5 different areas of Fars province during 2018 and 2019. DNAs of sand flies were extracted and PCR method was performed based on primers which were designed from surface proteins (WSP) genome region for Wolbachia and minicircle kDNA gene for Leishmania detections. At last, PCR products were sequenced and recorded in the GenBank. Out of 1002 sand flies caught from 5 different foci of Fars province, 909 male and female and 386 female sand flies' DNAs were extracted for detection of Wolbachia and Leishmania by PCR, respectively. Accordingly, out of the total 44 pools prepared from sand flies, 6 out of 28 pools form P. papatasi female were positive for Wolbachia in Shiraz, Marvdasht and Kharameh. However, none of female sand flies were positive for Leishmania. The study also focused on monitoring of sand flies co-infection of Wolbachia with Leishmania, which was not found in any of the studied samples. The negative results may be due to control strategies implemented which were done against Leishmaniasis in the studied areas during last years.