Evolution of MALDI-TOF MS Profiles from Lice and Fleas Preserved in Alcohol over Time.

MALDI-TOF is now considered a relevant tool for the identification of arthropods, including lice and fleas. However, the duration and conditions of storage, such as in ethanol, which is frequently used to preserve these ectoparasites, could impede their classification. The purpose of the present study was to assess the stability of MS profiles from Pediculus humanus corporis lice and Ctenocephalides felis fleas preserved in alcohol from one to four years and kinetically submitted to MALDI-TOF MS. A total of 469 cephalothoraxes from lice (n = 170) and fleas (n = 299) were tested. The reproducibility of the MS profiles was estimated based on the log score values (LSVs) obtained for query profiles compared to the reference profiles included in the MS database. Only MS spectra from P. humanus corporis and C. felis stored in alcohol for less than one year were included in the reference MS database. Approximately 75% of MS spectra from lice (75.2%, 94/125) and fleas (74.4%, 122/164) specimens stored in alcohol for 12 to 48 months, queried against the reference MS database, obtained relevant identification. An accurate analysis revealed a significant decrease in the proportion of identification for both species stored for more than 22 months in alcohol. It was hypothesized that incomplete drying was responsible for MS spectra variations. Then, 45 lice and 60 fleas were subjected to longer drying periods from 12 to 24 h. The increase in the drying period improved the proportion of relevant identification for lice (95%) and fleas (80%). This study highlighted that a correct rate of identification by MS could be obtained for lice and fleas preserved in alcohol for up to four years on the condition that the drying period was sufficiently long for accurate identification.

Molecular and MALDI-TOF MS characterisation of Hyalomma aegyptium ticks collected from turtles and their associated microorganisms in Algeria.

The identification of ticks and their associated pathogens is important for knowledge on tick-borne diseases. The objective of this study was to use morphological, matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) and/or molecular biology tools to identify ticks collected from turtles in north-eastern Algeria, as well as to investigate the microorganisms associated with these ticks. A total of 471 adult ticks were collected and identified morphologically as Hyalomma aegyptium, of which 248 (52.7%) were female and 223 (47.3%) were male. amongst them, 230 specimens were randomly selected for molecular and MALDI-TOF MS analysis. Molecular biology confirmed that our ticks were Hy. aegyptium. MALDI-TOF MS analysis revealed that 100% of the spectra were of excellent quality. Four spectra were selected to update our own database MALDI-TOF MS arthropod. The blind test of the 226 remaining spectra showed that all ticks were correctly identified, with scores ranging from 1.774 to 2.655 with a mean of 2.271 ± 0.16 of which, 223 (98.6%) had log score value (LSV)>1.8. Molecular biology screening showed that the ticks carried the DNA of Borrelia turcica, Rickettsia africae, Rickettsia aeschlimannii, Rickettsia sibirica mongolitimonae and with the Anaplasmataceae were close to a potentially new, undescribed Ehrlichia sp. This study confirms that MALDI-TOF MS is a reliable tool for the identification of ticks and that ticks collected from turtles in Algeria are carriers of several species of microorganisms which may be responsible for diseases in humans and animals.

Identification of Lice Stored in Alcohol Using MALDI-TOF MS.

Lice pose major public and veterinary health problems with economic consequences. Their identification is essential and requires the development of an innovative strategy. MALDI-TOF MS has recently been proposed as a quick, inexpensive, and accurate tool for the identification of arthropods. Alcohol is one of the most frequently used storage methods and makes it possible to store samples for long periods at room temperature. Several recent studies have reported that alcohol alters protein profiles resulting from MS analysis. After preliminary studies on frozen lice, the purpose of this research was to evaluate the influence of alcohol preservation on the accuracy of lice identification by MALDI-TOF MS. To this end, lice stored in alcohol for variable periods were submitted for MS analysis and sample preparation protocols were optimized. The reproducibility and specificity of the MS spectra obtained on both these arthropod families allowed us to implement the reference MS spectra database (DB) with protein profiles of seven lice species stored in alcohol. Blind tests revealed a correct identification of 93.9% of Pediculus humanus corporis (Linnaeus, 1758) and 98.4% of the other lice species collected in the field. This study demonstrated that MALDI-TOF MS could be successfully used for the identification of lice stored in alcohol for different lengths of time.