Currently microorganisms are best identified using 16S rRNA and 18S rRNA gene sequencing. However, in recent years matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has emerged as a potential tool for microbial identification and diagnosis. During the MALDI-TOF MS process, microbes are identified using either intact cells or cell extracts. The process is rapid, sensitive, and economical in terms of both labor and costs involved. The technology has been readily imbibed by microbiologists who have reported usage of MALDI-TOF MS for a number of purposes like, microbial identification and strain typing, epidemiological studies, detection of biological warfare agents, detection of water- and food-borne pathogens, detection of antibiotic resistance and detection of blood and urinary tract pathogens etc. The limitation of the technology is that identification of new isolates is possible only if the spectral database contains peptide mass fingerprints of the type strains of specific genera/species/subspecies/strains. This review provides an overview of the status and recent applications of mass spectrometry for microbial identification. It also explores the usefulness of this exciting new technology for diagnosis of diseases caused by bacteria, viruses, and fungi.

MALDI-TOF mass spectrometry: an emerging technology for microbial identification and diagnosis.

This review, focusing on studies published between 2005 and 2017, analysed the literature on the generation of bioactive peptides (BAPs) from edible insect proteins following enzymatic hydrolysis. The protein extraction and quantification methodologies used for edible insects varied considerably. While several edible insects have been evaluated for their ability to release BAPs, silkworm ( Bombyx mori ) is currently the most studied. Specifically, the angiotensin converting enzyme (ACE) inhibitory, antioxidant and antidiabetic properties of edible insect protein enzymatic hydrolysates have been studied. Potent in vitro ACE inhibitory and antioxidant hydrolysates/peptides have been reported. In certain instances, these properties were validated in small animal studies (i.e. hypotensive effects). Enzymatic hydrolysis of edible insect proteins may also enhance technofunctional properties (i.e. solubility). The wider application of enzymatic hydrolysis protocols to edible insect proteins may ultimately allow for the increased discovery and utilisation of novel BAPs as sustainable protein/peptide sources for human nutrition.

Unlocking the biological potential of proteins from edible insects through enzymatic hydrolysis: A review

The rapid and reliable identification of arthropod vector species is an essential component of the fight against vector-borne diseases. However, owing to the lack of entomological expertise required for the morphological identification method, development of alternative and complementary tools is needed. This review describes the main methods used for arthropod identification, focusing on the emergence of protein profiling using MALDI-TOF MS technology. Sample preparation, analysis of reproducibility, database creation and blind tests for controlling accuracy of this tool for arthropod identification are described. The advantages and limitations of the MALDI-TOF MS method are illustrated by emphasizing different hematophagous arthropods, including mosquitoes and ticks, the top two main vectors of infectious diseases.

Emerging tools for identification of arthropod vectors.

Over the past three decades, a range of mosquito-borne viruses that threaten public and veterinary health have emerged or re-emerged in Europe. Mosquito surveillance activities have highlighted the Culex pipiens species complex as being critical for the maintenance of a number of these viruses. This species complex contains morphologically similar forms that exhibit variation in phenotypes that can influence the probability of virus transmission. Critical amongst these is the choice of host on which to feed, with different forms showing different feeding preferences. This influences the ability of the mosquito to vector viruses and facilitate transmission of viruses to humans and domestic animals. Biases towards blood-feeding on avian or mammalian hosts have been demonstrated for different Cx. pipiens ecoforms and emerging evidence of hybrid populations across Europe adds another level of complexity to virus transmission. A range of molecular methods based on DNA have been developed to enable discrimination between morphologically indistinguishable forms, although this remains an active area of research. This review provides a comprehensive overview of developments in the understanding of the ecology, behaviour and genetics of Cx. pipiens in Europe, and how this influences arbovirus transmission.

The Role of Culex pipiens L. (Diptera: Culicidae) in Virus Transmission in Europe

Ticks are considered the second vector of human and animal diseases after mosquitoes. Therefore, identification of ticks and associated pathogens is an important step in the management of these vectors. In recent years, Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been reported as a promising method for the identification of arthropods including ticks. The objective of this study was to improve the conditions for the preparation of tick samples for their identification by MALDI-TOF MS from field-collected ethanol-stored Malian samples and to evaluate the capacity of this technology to distinguish infected and uninfected ticks. A total of 1,333 ticks were collected from mammals in three distinct sites from Mali. Morphological identification allowed classification of ticks into 6 species including Amblyomma variegatum, Hyalomma truncatum, Hyalomma marginatum rufipes, Rhipicephalus (Boophilus) microplus, Rhipicephalus evertsi evertsi and Rhipicephalus sanguineus sl. Among those, 471 ticks were randomly selected for molecular and proteomic analyses. Tick legs submitted to MALDI-TOF MS revealed a concordant morpho/molecular identification of 99.6%. The inclusion in our MALDI-TOF MS arthropod database of MS reference spectra from ethanol-preserved tick leg specimens was required to obtain reliable identification. When tested by molecular tools, 76.6%, 37.6%, 20.8% and 1.1% of the specimens tested were positive for Rickettsia spp., Coxiella burnetii, Anaplasmataceae and Borrelia spp., respectively. These results support the fact that MALDI-TOF is a reliable tool for the identification of ticks conserved in alcohol and enhances knowledge about the diversity of tick species and pathogens transmitted by ticks circulating in Mali.

/pdf/molecular-and-maldi-tof-identification-of-ticks-and-tick-3lau2vzt83.pdf

Molecular and MALDI-TOF identification of ticks and tick-associated bacteria in Mali.

MALDI-TOF MS profiling has proved to be efficient for arthropod identification at the species level. However, prior to entomological monitoring, the reference spectra database should cover relevant species. Here, 74 specimens were field-collected from 11 mosquito species captured in two distinct European areas and used either to increment our database or for blind tests. Misidentification was not noted, underlining the power of this approach. Nevertheless, three out of the 26 specimens used for the blind test did not reach the significant identification threshold value set, attributed to lower spectral quality. In the future, the quality control spectra parameters need to be defined to avoid not achieving significant threshold identification.

Identification of European mosquito species by MALDI-TOF MS.

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Identification of European mosquito species by MALDI-TOF MS.