Identification Of Specimens Research Articles

Culicoides species community composition and feeding preferences in two aquatic ecosystems in northern Spain

BackgroundAquatic ecosystems provide breeding sites for blood-sucking insects such as Culicoides biting midges (Diptera: Ceratopogonidae), but factors affecting their distribution and host choice are poorly understood. A study was undertaken at two nature reserves in northern Spain to examine the abundance, species composition, population dynamics and feeding patterns of biting midges between 2018 and 2019.MethodsCulicoides were captured by light suction traps baited with CO2 and by sweep netting vegetation. Blood meals and species identification of blood-fed specimens were determined using cytochrome c oxidase I subunit (COI) DNA barcoding. Multivariate generalized linear models were used to evaluate the associations between the abundance of Culicoides, the species richness and other parameters.ResultsThe 4973 identified specimens comprised 28 species of Culicoides. These included two species reported for the first time in northern Spain, thus raising to 54 the number of Culicoides species described in the region. Specimens of all 28 species and 99.6% of the total specimens collected were caught in suction traps, while sweep netting vegetation revealed just 11 species and 0.4% of the total specimens. Midge abundance peaked in June/early July, with five species comprising > 80% of the captures: Culicoides alazanicus (24.9%), Culicoides griseidorsum (20.3%), Culicoides poperinghensis (16.2%), Culicoides kibunensis (10.7%) and Culicoides clastrieri (9.6%). DNA barcode analysis of blood meals from eight Culicoides species revealed that they fed on 17 vertebrate species (3 mammals and 14 birds). Species in the subgenus Avaritia were primarily ornithophilic, except for C. griseidorsum and C. poperinghensis. Host DNA from blood meals was successfully amplified from 75% of blood-fed females. A pictorial blood meal digestion scale is provided to accurately assess the blood-fed status of female Culicoides.ConclusionsThe large number of different blood meal sources identified in the midges captured in this study signals the likely importance of wild birds and mammals (e.g. red deer and wild boar) as reservoir/amplifying hosts for pathogens. Available hosts are more exposed to being bitten by biting midge populations in aquatic ecosystems in late spring and early summer.

Lab Automation Based on IOT

Abstract: The concept of laboratory automation has existed for years; such automation has been used primarily in nonclinical and industrial settings. Specimen movement and result reporting are based on the identification of specimens using bar coded specimens and bar coded specimen carriers. The implementation of a laboratory automation system is dependent on the presence of a laboratory information system. An interface between the laboratory information system and the laboratory automation system provides the information required to move the specimen through the laboratory. The reporting of results is dependent on the laboratory information system or manual input, depending on the type of work cell in which the results are produced. The greatest hurdle to overcome in developing and implementing a laboratory automation system is the integration of systems, including commercial laboratory instrumentation and user-defined work cells. The barriers to implementation primarily are proprietary in nature: instrument software and instrument hardware. When the instrument manufacturers realize the necessity for development of electronic and physical integration, the proliferation of laboratory automation systems will occur. This article describes the author's concepts of laboratory automation.

A sequential decoding procedure for pooled quantitative measure

Group testing (pooling) is a commonly used method while screening for rare diseases, especially for large-scale screening. In recent years there has been growing interest in direct analysis of quantitative measures to take advantage of full information. In this study, we develop a pooling procedure for identification of infected specimens and the measurement is quantitative. Exclusive simulations are conducted to investigate the performance of our procedure, “Qpool,” under a variety of scenarios. We also demonstrate its application in routine monitoring of Listeria monocytogenes.

Development of a system for the automated identification of herbarium specimens with high accuracy

Herbarium specimens are dried plants mounted onto paper. They are used by a limited number of researchers, such as plant taxonomists, as a source of information on morphology and distribution. Recently, digitised herbarium specimens have begun to be used in comprehensive research to address broader issues. However, some specimens have been misidentified, and if used, there is a risk of drawing incorrect conclusions. In this study, we successfully developed a system for identifying taxon names with high accuracy using an image recognition system. We developed a system with an accuracy of 96.4% using 500,554 specimen images of 2171 plant taxa (2064 species, 9 subspecies, 88 varieties, and 10 forms in 192 families) that grow in Japan. We clarified where the artificial intelligence is looking to make decisions, and which taxa is being misidentified. As the system can be applied to digitalised images worldwide, it is useful for selecting and correcting misidentified herbarium specimens.

Forensic entomology in research and practice: an overview of forensic experts' perceptions and scientific output in Brazil.

Given the growing demands for standardization of procedures for collection, preservation, and identification of insect specimens, this study aimed to identify how forensic entomology is used in Brazil and the perceptions of professionals involved in this process. We conducted: (1) bibliometric survey of the scientific production of forensic specialists from 2001 to 2020; (2) questionnaire applied to forensic professionals to verify their perceptions about the use of entomological evidence in practice. An increase in publications on Forensic Entomology was noticed in the last 5years, mainly in the South and Northeast regions. Of the 82 respondents to the questionnaire, 62.8% deal with cadavers at least once a week, but 89.0% of the professionals who examine cadavers weekly find less than 10.0% colonized by insect larvae. Using entomological evidence to estimate PMI was mentioned by only 36.6% of the professionals. Most respondents stated no procedures related to collecting, handling, or identifying necrophagous insects in their daily practice. Regarding the chain of custody, 64.6% said they were aware of the changes in the regulations involving the collection and maintenance of forensic evidence. We emphasize the importance of collaboration between scientists at universities and crime scene examiners. Although the use of entomological evidence in criminal investigations in Brazil is still scarce, a growing interest in this area can be detected among forensic experts.

Light and scanning electron microscopic comparative studies of geminivirus infected and healthy Eclipta alba (L.).

Eclipta alba (L.) is a valuable medicinal plant. However, its medicinal efficacy can be affected by geminivirus infection. Therefore, identification of healthy specimen is essential before to use as medicine. The present study provided the taxonomic characterization of geminivirus infected and healthy E. alba plant by studying apparent morphology and microscopic features through light and scanning electron microscopy. Before taxonomic characterization infected and healthy specimens were separated through molecular detection of geminivirus. Results of morphological studies reported that geminivirus infected E. alba plant showed systematic symptoms of infection like stunted growth, distortion and chlorosis of leaves, decrease in size of root, shoot and fruit, and so forth in comparison to healthy specimen. Anatomical findings reported that in both plants anomocytic and anisocytic types of stomata with multicellular warty trichomes were present. However, variations were observed in quantitative measures such as size of trichomes, epidermal, subsidiary and guard cells. Palynological observations identifies that both plants possessed tricolporate type of pollen but variation was mainly observed in size and shape of pollen, thickness of exine and intine, P/E ratio, pore size, interspecific difference, size of colpi, and pollen ornamentation. Overall this study concluded that both healthy and infected E. alba do not reported much variations in qualitative taxonomic features, but can be differentiated in terms of quantitative taxonomic evidences. Future studies are recommended for pharmacological analysis of both healthy and virus infected plants. RESEARCH HIGHLIGHTS: Eclipta alba has incredible therapeutic worth, but due to geminivirus infection the plant is affecting badly. Hence, the present studies give a comprehensive taxonomic report on the geminivirus infected and healthy plant species.

Water eDNA metabarcoding is effective in detecting non-native species in marinas, but detection errors still hinder its use for passive monitoring

Marinas are high-priority targets for marine non-indigenous species (NIS), where they compose a large portion of the biofouling communities. The practicality of water samples collection makes environmental DNA (eDNA) metabarcoding an interesting tool for routine NIS surveys. Here the effectiveness of water-eDNA-metabarcoding to identify biofouling NIS, in 10 marinas from western France, was examined. Morphological identification of specimens collected in quadrats brought out 18 sessile benthic NIS beneath floating pontoons. Water-eDNA-metabarcoding detected two thirds of them, failing to detect important NIS. However, sampling and bioinformatics filtering steps can be optimized to identify more species. In addition, this method allowed the detection of additional NIS from neighboring micro-habitats. Caution should, however, be taken when reporting putative novel NIS, because of errors in species assignment. This work highlights that water-eDNA-metabarcoding is effective for active (targeted) NIS surveys and could be significantly improved for its further use in marine NIS passive surveys.

Organellar microcapture to extract nuclear and plastid DNA from recalcitrant wood specimens and trace evidence

BackgroundIllegal logging is a global crisis with significant environmental, economic, and social consequences. Efforts to combat it call for forensic methods to determine species identity, provenance, and individual identification of wood specimens throughout the forest products supply chain. DNA-based methodologies are the only tools with the potential to answer all three questions and the only ones that can be calibrated “non-destructively” by using leaves or other plant tissue and take advantage of publicly available DNA sequence databases. Despite the potential that DNA-based methods represent for wood forensics, low DNA yield from wood remains a limiting factor because, when compared to other plant tissues, wood has few living DNA-containing cells at functional maturity, it often has PCR-inhibiting extractives, and industrial processing of wood degrades DNA. To overcome these limitations, we developed a technique—organellar microcapture—to mechanically isolate intact nuclei and plastids from wood for subsequent DNA extraction, amplification, and sequencing.ResultsHere we demonstrate organellar microcapture wherein we remove individual nuclei from parenchyma cells in wood (fresh and aged) and leaves of Carya ovata and Tilia americana, amyloplasts from Carya wood, and chloroplasts from kale (Brassica sp.) leaf midribs. ITS (773 bp), ITS1 (350 bp), ITS2 (450 bp), and rbcL (620 bp) were amplified via polymerase chain reaction, sequenced, and heuristic searches against the NCBI database were used to confirm that recovered DNA corresponded to each taxon.ConclusionOrganellar microcapture, while too labor-intensive for routine extraction of many specimens, successfully recovered intact nuclei from wood samples collected more than sixty-five years ago, plastids from fresh sapwood and leaves, and presents great potential for DNA extraction from recalcitrant plant samples such as tissues rich in secondary metabolites, old specimens (archaeological, herbarium, and xylarium specimens), or trace evidence previously considered too small for analysis.

Rapid in situ identification of biological specimens via DNA amplicon sequencing using miniaturized laboratory equipment.

In many parts of the world, human-mediated environmental change is depleting biodiversity faster than it can be characterized, while invasive species cause agricultural damage, threaten human health and disrupt native habitats. Consequently, the application of effective approaches for rapid surveillance and identification of biological specimens is increasingly important to inform conservation and biosurveillance efforts. Taxonomic assignments have been greatly advanced using sequence-based applications, such as DNA barcoding, a diagnostic technique that utilizes PCR and DNA sequence analysis of standardized genetic regions. However, in many biodiversity hotspots, endeavors are often hindered by a lack of laboratory infrastructure, funding for biodiversity research and restrictions on the transport of biological samples. A promising development is the advent of low-cost, miniaturized scientific equipment. Such tools can be assembled into functional laboratories to carry out genetic analyses in situ, at local institutions, field stations or classrooms. Here, we outline the steps required to perform amplicon sequencing applications, from DNA isolation to nanopore sequencing and downstream data analysis, all of which can be conducted outside of a conventional laboratory environment using miniaturized scientific equipment, without reliance on Internet connectivity. Depending on sample type, the protocol (from DNA extraction to full bioinformatic analyses) can be completed within 10 h, and with appropriate quality controls can be used for diagnostic identification of samples independent of core genomic facilities that are required for alternative methods.

DNA barcodes in identification of some species of hawkmoths (Lepidoptera: Sphingidae)

The use of DNA barcodes is a molecular method commonly used for the identification of specimens that are difficult to pinpoint accurately by traditional morphological methods. DNA barcoding uses standard short genomic regions that are universally present in target lineages and has sufficient sequence variation to identify species in the genus. The hawkmoth genus Cechetra (Zolotuhin & Ryabov, 2012) was proposed for a group of morphologically similar, stripe-patterned species. Based on the seven specimens of the three species of Cechetra genus collected from the Central of Vietnam, we used morphological analysis to initially identified their scientific name of Cechetra lineosa (Walker, 1856) for 04 specimens (LB_F01, LB_G01, LB_H01 and LK_G05); 02 specimens (MK_H04 and MK_B07) were identified with the scientific name of Cechetra minor (Butler, 1875) and 01 specimen (SG_D7) with scientific name Cechetra subangustata (Rothschild, 1920). Then, the COI (Cytochrome C oxidase I) sequence was used for molecular analysis. Total DNA was extracted from the legs of specimens, COI with over 600 bp in length from each specimen was amplified by PCR reactions. The PCR products are hence purified and sequenced. COI gene cloning results showed a very high level of genetic similarity (over 99%) and 7 specimens are of the genus Cechetra, Sphingidae family. For three homomorphic species belonging to the genus Cechetra Zolotuhin & Ryabov, 2012 including Cechetra lineosa (Walker, 1856); Cechetra minor (Butler, 1875) and Cechetra subangustata (Rothschild, 1920). The studied sequences have been added to the current database at GenBank (NCBI) with accession numbers MT994230, MT994231, MT994232, MT994233, MT994234, MT994235, and MT994236.

Sorting the flock: Quantitative identification of sheep and goat from isolated third lower molars and mandibles through geometric morphometrics

Sheep and goat are often herded together and show morphological similarities in their skeleton. Being able to identify archaeological remains of these two taxa to species level is particularly important for understanding and characterising past herding practices. Discrete criteria are now available to identify a large number of their bones and teeth, and quantitative approaches have been developed for post-cranial elements but not for mandible and isolated teeth. In this paper we explore the discriminating potential of geometric morphometrics to identify modern sheep and goat third lower molar and mandible and its application on archaeological specimens. The size and shape of the mandible and the third lower molar of 143 modern specimens (101 sheep and 42 goats) were quantified using 2D-landmark and sliding semi-landmarks geometric morphometric approaches. The results show that sheep and goat differ in terms of the size, shape, and form (i.e. size and shape together) in both studied elements. Classification accuracy of the two species reaches 93.3% (CI: 90.0–95.7%) for third lower molar shape, 62.7% (CI 57.1–68.6%) for third lower molar size, 95.2% (CI: 92.0–97.4%) for mandible shape and 84.0% (CI 81.6–86.8%) for mandible size. Form does not provide better classification than shape alone. Sex and age appear to have little impact on the ability to differentiate between sheep and goat, despite the two species displaying distinct sexual dimorphism and changes through age. The same methodology was then applied on 32 Middle Ages third lower molars from Missignac-Saint Gilles le Vieux, Aimargues, France. The identifications obtained through geometric morphometrics were only partially congruent with the identifications based on visual observations calling for caution in the interpretation and further investigations. Further research should include molecular identification of the archaeological specimens to assess whether the geometric morphometric identification can be made with confidence for all periods and all geographic areas. Nevertheless, the results obtained with the newly developed geometric morphometric protocols represent an important contribution toward a better understanding of past livestock husbandry practices.

From alchemy to modern mineralogy: dating mineral collections via chemical notation

Mineral specimens found in historical collections often include specimen labels, which may provide vital information on the nature, chemistry and origin of the material. However, the evolution of chemical notation, particularly during the eighteenth and nineteenth centuries, led to a wide range of ways in which a given sample could be adequately documented, many of which may still be found in collections in the present day. Prior to the advent of modern mineralogy, samples were labelled using a complex and sometimes baffling language of alchemical symbols with little meaning to modern scientists. The efforts of notable chemists and mineralogists, such as Torbern Bergman, Antoine Lavoisier, John Dalton and Jöns Jacob Berzelius, began to reform and unify chemical nomenclature, providing a number of new terms, symbols and approaches to the description of materials. With the advent of atomic theory in the nineteenth century, new classification schemes based on quantitatively describing the atomic arrangement and composition of minerals were proposed, representing a significant step towards modern mineralogy. Understanding the development of chemical notation over time not only facilitates the identification of mineral specimens but may also provide clues as to the date that the sample was documented, and potentially even the location.

Darwin wasps (Hymenoptera, Ichneumonidae) in Lower Eocene amber from the Paris basin

Despite their ecological importance, Darwin wasps (Ichneumonidae) are among the most poorly studied groups of organisms. It is therefore not surprising that their fossil record is even more poorly understood than their extant diversity. The early Eocene seems rather fossil-poor regarding Ichneumonidae in amber and only one species,Palaeometopius eocenicusMenier et al., 2004, was described so far from Oise Amber from the Paris basin. Here, two new ichneumonid genera and species,Madma oisellagen. et. sp. nov.andPappous trichomatiusgen. et. sp. nov.are described and the placement ofPalaeometopius eocenicusis revised. The three fossils are well-preserved and might represent stem taxa of Tryphoninae and Phygadeuontinae. They are a highly important addition to the early Palaeogene fossil record of Ichneumonidae that otherwise mainly consists of compression fossils, which yield far less detail of the specimens’ morphology than amber pieces. Among the more than 1,000 Oise amber pieces examined, only three Ichneumonidae specimens have been found, versus about 60 Braconidae, a ratio very different from other amber deposits. Identification of additional ichneumonid specimens from this period (lowermost Eocene) is of particular importance for a better understanding of the subfamily and species compositions of this family after the K-Pg mass extinction.

Modified Mosquito Programs’ Surveillance Needs and An Image-Based Identification Tool to Address Them

Effective mosquito surveillance and control relies on rapid and accurate identification of mosquito vectors and confounding sympatric species. As adoption of modified mosquito (MM) control techniques has increased, the value of monitoring the success of interventions has gained recognition and has pushed the field away from traditional ‘spray and pray’ approaches. Field evaluation and monitoring of MM control techniques that target specific species require massive volumes of surveillance data involving species-level identifications. However, traditional surveillance methods remain time and labor-intensive, requiring highly trained, experienced personnel. Health districts often lack the resources needed to collect essential data, and conventional entomological species identification involves a significant learning curve to produce consistent high accuracy data. These needs led us to develop MosID: a device that allows for high-accuracy mosquito species identification to enhance capability and capacity of mosquito surveillance programs. The device features high-resolution optics and enables batch image capture and species identification of mosquito specimens using computer vision. While development is ongoing, we share an update on key metrics of the MosID system. The identification algorithm, tested internally across 16 species, achieved 98.4 ± 0.6% % macro F1-score on a dataset of known species, unknown species used in training, and species reserved for testing (species, specimens respectively: 12, 1302; 12, 603; 7, 222). Preliminary user testing showed specimens were processed with MosID at a rate ranging from 181-600 specimens per hour. We also discuss other metrics within technical scope, such as mosquito sex and fluorescence detection, that may further support MM programs.

Evaluation of human melanoma and normal formalin paraffin-fixed samples using Raman and LIBS fused data.

In this research, we developed a novel method of quantitative analysis to increase the detection potential for screening and classification of skin cancer (melanoma). We fused two distinct optical approaches, an atomic spectroscopic detection technique laser-induced breakdown spectroscopy (LIBS) and a vibrational molecular spectroscopic technique known as Raman spectroscopy. Melanoma is a kind of skin cancer, also known as malignant melanoma, that developed in melanocytes cells, which produced melanin. Classification of melanoma cancerous tissues is a fundamental problem in biomedicine. For early melanoma cancer diagnosis and treatment, precise and accurate categorizing is critically essential. Laser-based spectroscopic approaches can be used as an operating instrument for simultaneous tissue ablation and ablated tissue elemental and molecular analysis. For this purpose, melanoma and normal paraffin-embedded tissues are used as a sample for LIBS and Raman measurement. We studied the data provided by laser-based spectroscopic methods using different machine learning classification techniques of extreme learning machine (ELM), partial least square discriminant analysis (PLS-DA), and K nearest neighbors (kNN). For visualization of melanoma and normal data, principal component analysis (PCA) is also used. Three different ways are used to process the data, LIBS measurement, Raman measurement, and combine data measurement (merged/fused data), and then compared the results. ELM classification model achieved the highest accuracy (100%) for combined data as well as for Raman and LIBS data, respectively. According to the experimental results, we can assume that Raman spectroscopy and LIBS combine can significantly improve the identification and classification accuracy of melanoma and normal specimens.

First evidence of White-footed Deer Mouse (<i>Peromyscus leucopus</i>) on mainland New Brunswick, Canada

White-footed Deer Mouse (Peromyscus leucopus) and the closely related, and more northerly ranging, Deer Mouse (Peromyscus maniculatus) broadly overlap in distribution and are often difficult to distinguish from each other. Based on molecular genetic data (cytochrome b gene), we report two new distribution records for P. leucopus for New Brunswick, Canada, the first mainland localities for this species in the province. Previous sampling of Peromyscus in New Brunswick may have overlooked the presence of P. leucopus, possibly because the specimens collected were all assumed to be P. maniculatus. However, current detection in New Brunswick may be part of a broader recent northward range expansion documented to be underway in P. leucopus. Although our use of a single mitochondrial gene to identify P. leucopus does not eliminate the possibility that the New Brunswick specimens are of hybrid origin, our results support the presence of P. leucopus in New Brunswick and suggest more detailed analyses will be required to determine the nature of any genetic interaction between P. leucopus and P. maniculatus in the province. Recognition of morphologically cryptic Peromyscus in southern New Brunswick also emphasizes the need to incorporate comprehensive methods to ensure the correct identification of specimens of this genus in Maritime Canada. We also note the potential implications of this discovery with respect to the incidence of Lyme disease in New Brunswick.

Fast and novel botanical exploration of a 320-km transect in eastern Amazonia using DNA barcoding

ABSTRACT We explored a 320-km transect in the Tumucumaque mountain range along the border between southern French Guiana and Brazil, sampling all trees and lianas with DBH ≥ 10 cm in seven 25 x 25-m plots installed near seven boundary milestones. We isolated DNA from cambium tissue and sequenced two DNA barcodes (rbcLa and matK) to aid in species identification. We also collected fertile herbarium specimens from other species (trees/shrubs/herbs) inside and outside the plots. The selected DNA barcodes were useful at the family level but failed to identify specimens at the species level. Based on DNA barcoding identification, the most abundant families in the plots were Burseraceae, Fabaceae, Meliaceae, Moraceae, Myristicaceae and Sapotaceae. One third of the images of sampled plants posted on the iNaturalist website were identified by the community to species level. New approaches, including the sequencing of the ITS region and fast evolving DNA plastid regions, remain to be tested for their utility in the identification of specimens at lower taxonomic levels in floristic inventories in the Amazon region.

SPECIES RICHNESS OF MACROLICHENS IN MT. SAPINIT, SITIO DAGULOS, LA ROXAS, MARAMAG, BUKIDNON

The species richness of macrolichens in Mt. Sapinit, SitioDagulos, and MaramagBukidnon was investigated. The main objective of the study is to determine the species richness of macrolichens in the vicinity of Mt. Sapinit. Transect walk and quadrat sampling were carried out in the collection of macrolichens. The gathering and identification of specimens collected was based on standard taxonomical procedure. Results of the study revealed a total of 20 species classified into 7 genera and 13 families. The most family-rich species belong to the family Lobariaceae, and the least family-rich belong to Ramalinaceae, Collemataceae, and Dictyonemaceae. Most of the epiphytic macrolichens belong to the family Parmeliaceae, which dominates the entire montane forest. The distribution of macrolichens across the area occurs in the bark of the tree, logs and twigs, and in tree branches.Elevation also have impact to the growth of lichen species. Higher elevations appear to provide suitable condition for many lichen species to thrive. Thus, the need to preserve the forest ecosystem is essential in order to conserve the lichen communities.

High-Throughput Sequencing for Life-History Sorting and for Bridging Reference Sequences in Marine Gerromorpha (Insecta: Heteroptera)

AbstractAccurate identification and association of larval specimens with adults is a major challenge in insect taxonomy. Fortunately, it is now possible for nonexperts to sort collections of bulk samples with DNA barcodes rapidly and cost-effectively. We demonstrate this process using nanopore barcoding of 757 marine insects (Insecta: Gerromorpha), of which 81% were nymphs and many samples did not have co-occurring adult males for specific identification. We successfully associated 738 specimens (97%) to nine gerromorphan species, which would have been impossible to identify using morphological characters alone. This improved ability to incorporate information from all life-history stages has led to greater precision of species distributional ranges—knowledge that will be crucial for a more complete understanding of marine insects. We also highlighted two distinct, nonoverlapping Gerromorpha COI sequence databases on GenBank—a consequence of using two different primer sets to amplify different regions of COI. This issue inevitably hinders species identification with DNA-based methods, particularly for poorly represented groups such as marine insects. We bridged these databases by analyzing full-length COI sequences. We believe this will inspire future studies to incorporate DNA-based methods for more adult–larval association studies and for enhancing existing genetic resources, especially in understudied groups.

Parasitism, seasonality, and diversity of trombiculid mites (Trombidiformes: Parasitengona, Trombiculidae) infesting bats (Chiroptera) in Poland

The study aims to ascertain the diversity of trombiculid species associated with Chiroptera in Poland, and for the first time in the case of research on Central European Trombiculidae, we use both DNA and morphology in an integrative taxonomic approach to determine species identities of trombiculids. The research was carried out from 2015 to 2019. In total, 2725 larvae were collected from 300 specimens of bats belonging to 11 species. Deutonymphs were obtained through laboratory rearing of larvae; few larvae and deutonymphs were collected also from bats' daily roosts. The presence of trombiculid larvae on hosts was observed between July and April of the following year, with the highest numbers recorded in autumn, during bat swarming. Male bats were infested more often than females (16.4 vs. 6.6%). The highest infestation rate was recorded for Barbastella barbastellus, Myotis nattereri and Plecotus auritus, and the highest prevalence of chiggers (> 30%) for Myotis bechsteinii and P. auritus. The larvae found on bats occupied the areas with free access to the host’s skin: auricles, tragus, and snout. Morphological identification of specimens to the species level was hindered by the mosaic distribution of diagnostic traits. Morphological analyses indicated the presence of Leptotrombidium russicum and Leptotrombidium spp. in the examined material, whereas molecular analyses additionally suggested three other potential species assigned to the same genus based on the assessed scope of intrageneric variation (ASAP method). We argue that the identification of the parasitic larvae (chiggers) using morphological characters does not address the question of actual species boundaries, which, in turn, affects the inferences about host specificity and host range.