Issue : 01
INTERNATIONAL
JOURNAL
OF
DIAGNOSTICS
AND
RESEARCH
Volume : 04
Copyright @ : - Dr.Laman Dadashova Inter. J.Digno. and Research IJDRMSID0144 |ISSN :2584-2757
46
P
ISSN No. : 2584-2757
Volume : 04
Issue : 01
DOI
:10.5281/zenodo.23434757
Reg. No. : MAHA-703/16(NAG)
Year of Establishment – 2016
INTERNATIONAL JOURNAL OF DIAGNOSTICS AND RESEARCH
Corresponding Author: Dr. Laman Dadashova
ORCID ID: 0009-0004-2966-2523
ISI Impact Factor (2025-26): 1.345
IIFS Impact Factor (2026-27): 6.0
Article Info: Article Received on : 05/08/2026 Article Reviewed on: 12/09/2026 Article Published on : 15/10/2026
Cite this article as: - Dadashova L. Molecular-Genetic Characteristics Of Representatives Of The Genus Hirudo: A Review Of
Modern Studies. International Journal of Diagnostics And Research. 2026Oct17;4(1):46–55. doi:10.5281/zenodo.23434757
Abstract
The bite of an in infected Aedes mosquito (Aedes aegypti or, Aedes albopictus) can transmit the virus that
This review summarizes and analyzes recent molecular genetic studies on the genus Hirudo, with particular
emphasis on Hirudo orientalis. The review is based primarily on scientific literature published between
2004 and 2024, covering molecular identification, genetic diversity, and phylogenetic relationships among
species. The analysis demonstrates that identification based solely on morphological characteristics is
insufficient for the accurate discrimination of morphologically similar species such as H. medicinalis, H.
verbana, and H. orientalis. The application of mitochondrial COI and 12S rRNA markers, together with
nuclear ITS1/ITS2 markers, has enabled the identification of genetic differences among these species and
supported their recognition as distinct taxa. In particular, molecular studies of H. orientalis have revealed
the presence of intraspecific genetic diversity and distinct haplotypes, while also contributing significantly
to the understanding of its geographic and phylogenetic characteristics. Overall, the application of
molecular genetic methods is of considerable importance for the accurate identification of Hirudo species,
determination of their taxonomic status, investigation of their phylogenetic relationships, and assessment of
the genetic diversity of their natural populationsviridae family, are transmitted by the bite of an infected
aedes mosquito. Dengue is widespread in over 100 nations worldwide. There is currently no specific
treatment for this illness in modern medicine. According to Ayurveda, Dandaka Jwara, is associated with
dengue fever. For this reason, it is often referred to as "break bone fever." Since contemporary science has
no cure for dengue fever, Ayurveda can be a very helpful tool in controlling the illness. However, a
complete understanding of dengue fever from both a modern and an ayurvedic perspective is required.
Keywords: Molecular, review article, Hirudo, Hirudo orientalis, phylogenetic.
Molecular-Genetic Characteristics Of Representatives Of The Genus Hirudo: A Review Of
Modern Studies
Dr. Laman Dadashova
1
1
Ph.D. Scholar ,Ministry of Education and Science of the Republic of Azerbaijan Institute of Living Systems
Research
G
A
R
V
Issue : 1
INTERNATIONAL JOURNAL OF DIAGNOSTICS AND RESEARCH [ISSN No.: 2584-2757]
Volume : 4
Copyright @ : - Dr.Laman Dadashova Inter. J.Digno. and Research IJDRMSID0144 |ISSN :2584-2757
47
Introduction :
Aquatic ecosystems worldwide are characterised by
a rich hydrofauna. However, these groups of
organisms face constant pressure from both natural
factors and anthropogenic impacts. Consequently,
continuous observation and monitoring of aquatic
organisms (hydrobionts) are important. A key
characteristic of aquatic ecosystems is their
interaction with the surrounding landscape and
biocenoses. In other words, changes in freshwater
bodies—such as pollution from industrial and
agricultural activities, the accumulation of heavy
metal-rich waste, excess organic matter, and
ecological disturbances linked to urbanisation—
directly influence the composition of resident
organisms, particularly the hirudofauna.
Seven main species belonging to the genus Hirudo
are widely used worldwide for medical purposes:
1. Hirudo medicinalis – the classic European
medicinal leech, distributed primarily across
Northern and Western Europe
[16]
2. Hirudo verbana – distributed across Southern
and Southeastern Europe; morphologically
similar to H. medicinalis, yet genetically
distinct
3. Hirudo orientalis – Distributed in the Caucasus,
Iran, and Central Asia; confirmed as a distinct
species through molecular studies
4. Hirudo troctina – Found primarily in North
Africa (Morocco, Algeria, Tunisia)
5. Hirudo sulukii – Discovered in Turkey (Aegean
and Mediterranean regions); described by N.
SaÄŸlam in 2016
6. Hirudo tianjinensis – Described as a new
species by Wang et al. in 2022 in the Tianjin
region of China
7. Hirudo nipponia – Although assigned to a
separate genus (Whitmania) by some authors, it
is also classified within Hirudo in certain
systematic approaches
Materials And Methods :
Cyclical glacial events have significantly shaped
the genetic divergence and biogeographical
distribution of certain species within the genus
Hirudo. Climatic fluctuations, particularly those
associated with the Pleistocene epoch, fragmented
populations into geographically isolated refugia,
promoting genetic differentiation and increasing
intraspecific genetic diversity.
Medicinal leeches have long been important in both
medicine and zoological systematics. The
nineteenth century saw the establishment of
fundamental knowledge about their diversity and
taxonomic classification through the pioneering
studies of Grube and Whitman. Research during
this period focused primarily on external
morphological characteristics, including similarities
and differences in body form and patterns of
geographical distribution. Although these
morphological approaches provided an important
foundation for early classification of medicinal
leeches, relying solely on phenotypic characteristics
proved insufficient for accurate taxonomic
discrimination and comprehensive classification
[16].
By the mid-twentieth century, studies conducted by
Epstein and Livanova had focused on the
identification and taxonomic
characterisation of leech species occurring
throughout the territory of the former USSR.
However, advances in molecular genetics led to
Issue : 1
INTERNATIONAL JOURNAL OF DIAGNOSTICS AND RESEARCH [ISSN No.: 2584-2757]
Volume : 4
Copyright @ : - Dr.Laman Dadashova Inter. J.Digno. and Research IJDRMSID0144 |ISSN :2584-2757
48
new approaches that overcame the limitations of
classical morphological methods. Molecular
genetic markers enabled more accurate delineation
of interspecific boundaries and provided more
reliable evidence for species identification and
taxonomic assessment. Consequently, molecular
approaches have become an essential complement
to traditional morphological analyses in the study
of leech diversity, taxonomy, and evolutionary
relationships
[5]
. For example, despite their
considerable morphological similarity, molecular
analyses have shown that H. medicinalis, H.
verbana, and H. orientalis represent distinct
species. Studies based on mitochondrial DNA
markers, particularly the cytochrome c oxidase
subunit I (COI) and 12S rRNA genes, as well as
nuclear DNA markers, including the ITS1 and ITS2
regions, have confirmed substantial genetic
differentiation among these taxa. These findings are
important for accurately identifying and
taxonomically distinguishing closely related Hirudo
species and for more precisely assessing their
biogeographical distribution patterns
[13]
.
Hybridisation experiments conducted by
PetrauskienÄ— and colleagues have demonstrated that
artificial cross-breeding among different species of
the genus Hirudo is possible under controlled
laboratory conditions. However, the results
revealed that the vast majority of hybrid offspring
exhibited reduced viability or were completely non-
viable. These findings provide further evidence of
reproductive isolation among the species concerned
and support their recognition and classification as
distinct taxonomic entities. Laboratory experiments
involving artificial hybridisation among three
species—H. medicinalis, H. verbana, and H.
orientalis—have provided evidence of reproductive
incompatibility between these taxa. In one
experimental cross, female H. medicinalis and male
H. verbana individuals produced cocoons after a
defined mating period. However, the offspring that
emerged from these cocoons were poorly
developed and exhibited markedly reduced
viability. Complete mortality of the offspring was
observed within eight months, resulting in a
reproductive success rate of zero. These findings
provide strong evidence of reproductive isolation
and genetic incompatibility between the species
involved, further supporting their recognition as
distinct taxonomic entities
[12]
.
Laboratory experiments involving artificial
hybridisation among three species—H. medicinalis,
H.verbana, and H.orientalis—have provided
evidence of reproductive incompatibility between
these taxa. In one experimental cross, female H.
medicinalis and male H. verbana individuals
produced cocoons after a defined mating period.
However, the offspring that emerged from these
cocoons were poorly developed and exhibited
markedly reduced viability. Complete mortality of
the offspring was observed within eight months,
resulting in a reproductive success rate of zero.
These findings provide strong evidence of
reproductive isolation and genetic incompatibility
between the species involved, further supporting
their recognition as distinct taxonomic entities.
Analysis of the hybridisation results provides
evidence for genetic barriers among Hirudo
species, supporting their recognition as distinct
species. In modern systematics, reproductive
Issue : 1
INTERNATIONAL JOURNAL OF DIAGNOSTICS AND RESEARCH [ISSN No.: 2584-2757]
Volume : 4
Copyright @ : - Dr.Laman Dadashova Inter. J.Digno. and Research IJDRMSID0144 |ISSN :2584-2757
49
isolation is a principal criterion for delineating
species boundaries. Molecular phylogenetic studies
by Siddall and Borda further show that, despite
their pronounced morphological similarity, H.
verbana and H. medicinalis represent genetically
distinct evolutionary lineages
[13]
.
Trontelj and Utevsky reported similar findings,
suggesting that the reduced viability observed in
hybridisation experiments was associated with
reproductive isolation. Molecular-genetic analyses
later corroborated the polymorphic traits described
in early taxonomic accounts of H. medicinalis.
Studies have shown that variation in colouration
and body patterning within this species is not
merely random phenotypic variation but is
associated with distinct morphotypes. Furthermore,
comparative analysis of cytochrome c oxidase
subunit I (COI) gene sequences available in the
GenBank database has revealed genetic
polymorphism in H. medicinalis. We applied the
same approach to H. verbana, showing that this
species also exhibits substantial genetic diversity.
Collectively, these findings indicate that, despite
their morphological similarities, molecular markers
can reveal considerable genetic variation within
both species
[17-18]
.
Asian species, particularly H. nipponia and H.
tianjinensis, differ from their European congeners
in both morphology and genetics. Molecular
analyses based on cytochrome c oxidase subunit I
(COI) gene sequences indicate that these species
are phylogenetically related to other members of
the genus Hirudo, while representing distinct
evolutionary lineages. These findings suggest that
geographic isolation may have driven their
evolutionary divergence, accompanied by
independent ecological adaptation
[8]
.
In recent studies, Hildebrandt and Lemke examined
the taxonomic relationships among the three most
widespread medicinal leech species in Europe:
Hirudo medicinalis Linnaeus, 1758, Hirudo
verbana Carena, 1820, and Hirudo orientalis
Utevsky & Trontelj, 2005
[7].
The authors noted that
although these species' geographical ranges may
overlap, their reproductive compatibility under
laboratory conditions remains poorly understood.
Reports of successful mating under artificial
conditions have raised questions regarding the
extent of reproductive isolation among these taxa.
However, successful mating alone does not
necessarily indicate reproductive compatibility;
further evidence is needed to determine whether
they can produce viable, fertile offspring.
Therefore, researchers should evaluate their
taxonomic status alongside morphological,
molecular-genetic, and reproductive evidence
[6]
.
However, these findings have prompted debate
among researchers regarding the extent of
reproductive isolation among medicinal leech
species. Studies by Elliott and Kutschera indicate
that interspecific crosses conducted under
laboratory conditions often produce offspring with
low viability and limited ability to survive long-
term. These findings suggest reproductive barriers
between these taxa and support their recognition as
distinct species. Nevertheless, reproductive
isolation should be assessed based on offspring
viability, fertility, and the ability to establish
subsequent generations, alongside molecular-
genetic and morphological evidence
[3]
.
Issue : 1
INTERNATIONAL JOURNAL OF DIAGNOSTICS AND RESEARCH [ISSN No.: 2584-2757]
Volume : 4
Copyright @ : - Dr.Laman Dadashova Inter. J.Digno. and Research IJDRMSID0144 |ISSN :2584-2757
50
Specimens of Hirudo collected from small ponds in
Hungary and characterised by an unpigmented,
greyish ventral surface were found to differ from
the original descriptions provided by Linnaeus.
Consequently, these specimens were assigned to
Hirudo verbana Carena, 1820, also associated in
the literature with the name H.officinalis Savigny,
1822. Subsequent studies demonstrated that,
despite their morphological similarities,
H.medicinalis and H.verbana represent genetically
distinct species. Although both species inhabit
freshwater environments, evidence of natural
interbreeding and offspring produced between them
has not been established. These findings prompted
more detailed morphological and taxonomic
investigations, alongside the determination and
comparative analysis of their mitochondrial DNA
sequences. The resulting evidence supported their
recognition as two distinct species
[10]
.
Research conducted by K. Darabi-Darestani and
colleagues revealed substantial haplotype diversity
in Hirudo orientalis populations from the Caspian
region. The authors suggested that prolonged
geographical isolation may explain the observed
differences in population genetic structure. The
authors also highlighted the role of historical
refugia, which may have facilitated the persistence
of isolated populations and promoted genetic
divergence over evolutionary timescales, as a
potential factor contributing to the emergence of
this genetic diversity
[1]
.
Such studies provide valuable insights not only into
taxonomic identification but also into evolutionary
biology and biogeography. Phylogenetic and
population-genetic data enable more accurate
reconstruction of Hirudo species' evolutionary
history and a better understanding of their
distribution patterns and dynamics
[2]
. The
GenBank database includes the COI gene sequence
for H. verbana var. verbana under accession
number EF125043. Hildebrandt and Lemke used
this molecular marker to investigate the taxonomic
identity of the specimens, and their results support
assignment to H. verbana. Comparative studies
have also examined phylogenetic relationships
among the three European medicinal leech species
and the North African species Hirudo troctina
Johnson, 1816, commonly known as the dragon
leech. These taxa have been discussed as part of a
closely related species group. Research by
Kutschera and other investigators has also
highlighted genetic polymorphism in H. verbana,
demonstrating distinct genetic variants within the
species
[7]
. Hirudo tianjinensis, described by
Chinese researchers, exhibits morphological and
genetic characteristics that distinguish it from other
members of the genus Hirudo. Its diagnostic
external features include five yellow longitudinal
stripes on the dorsal surface, dark-brown spots on
the ventral side, and a high density of sensory
organs. Molecular analyses of the cytochrome c
oxidase subunit I (COI) gene indicate a close
phylogenetic relationship between H. tianjinensis
and the Asian species H. nipponia Whitman, 1886.
We developed an optimised protocol for
recombinant production and purification of a novel
cysteine-rich anticoagulant (CRA) protein derived
from Hirudo medicinalis saliva. The yield of
purified CRA ranged from 3.7 to 5.5 mg/L, and we
assessed levels of potential contaminants, including
Issue : 1
INTERNATIONAL JOURNAL OF DIAGNOSTICS AND RESEARCH [ISSN No.: 2584-2757]
Volume : 4
Copyright @ : - Dr.Laman Dadashova Inter. J.Digno. and Research IJDRMSID0144 |ISSN :2584-2757
51
bacterial proteins, DNA, and endotoxins. The
purified protein was highly pure and a potent
anticoagulant. In contrast, a truncated CRA variant
lacking the C-terminal region lost its membrane-
binding capacity and showed an altered
anticoagulant profile, characterised by increased
thrombin time effect and reduced prothrombin time
and activated partial thromboplastin time effects.
These findings highlight CRA's potential as a
promising candidate for further investigation as an
anticoagulant agent.Accurate species identification
is essential for ensuring the species-specific
efficacy and quality of medicinal leeches used in
medical applications. However, conventional DNA
barcoding methods may be ineffective for
traditional medicinal preparations because DNA
degradation can compromise amplification and
analysis of standard barcode regions. This
limitation
underscores the need to develop mini-barcoding
approaches that target shorter DNA fragments,
thereby improving the molecular identification of
medicinal leech species in degraded biological
material.This study aimed to develop and validate
mini-barcode markers for the molecular
identification of three leech species listed in the
Chinese Pharmacopoeia: Whitmania pigra,
Whitmania acranulata, and Hirudo nipponia. We
designed and validated four novel mini-barcode
primer pairs (ND1F1/R1, 12SF1/R1, 16SF1/R1,
and COX1F1/R1) using seven morphologically
identified specimens. We then evaluated these
markers using 16 leech-derived preparations
[9]
.
We extracted DNA using two approaches: a single-
tube extraction method and column-based
purification kits. The column-based method yielded
higher-quality DNA suitable for subsequent
polymerase chain reaction (PCR)
amplification. PCR analysis confirmed that all four
candidate mini-barcode markers targeted specific
genomic regions and were applicable, with
successful amplification from 13 of the 16
preparations. Mini-barcode sequences from
morphologically identified W. pigra specimens
showed> 95% sequence identity to the
corresponding reference sequences for ND1, 12S
rDNA, 16S rDNA, and COX1 (GenBank accession
number EU304459). In contrast, sequence identity
for H. nipponia and W. acranulata specimens was
below 85%. Among the leech-derived preparations
examined, only the traditional Chinese medicinal
product Maxuekang exhibited comparatively lower
sequence similarity.
Optimal clustering and phylogenetic tree analyses
using the Automated Single-Cell Analysis Pipeline
(ASAP) identified three distinct clusters
corresponding to the morphologically identified
species: W. pigra, W. acranulata, and H. nipponia.
The analyses also revealed that specimens
previously identified as Hirudo nipponia in some
studies were, in fact, W. pigra, highlighting the
potential for taxonomic misidentification based on
morphology alone.Overall, the findings show that
mini-barcoding can substantially improve species-
level identification and support quality control of
traditional medicinal and commercially available
leech-derived products. This approach may support
safer, more reliable use of medicinal leeches and
the incorporation of molecular species-
identification procedures into regulatory and
Issue : 1
INTERNATIONAL JOURNAL OF DIAGNOSTICS AND RESEARCH [ISSN No.: 2584-2757]
Volume : 4
Copyright @ : - Dr.Laman Dadashova Inter. J.Digno. and Research IJDRMSID0144 |ISSN :2584-2757
52
quality-control standards. According to the
traditional taxonomic view, all morphological
forms of European medicinal leeches are assigned
to a single species, Hirudo medicinalis. However, a
A study using random amplified polymorphic DNA
(RAPD) markers showed that Hirudo verbana,
another species distributed in Europe, is a distinct
biological species. Genetic clustering and Principal
Coordinates Analysis (PCoA) of eight populations
revealed consistent genetic differentiation,
indicating that the observed structure was driven
primarily by taxonomic divergence rather than
geographic variation. Interspecific differences
accounted for 60% of the total molecular variation
(φCT = 0.60; P < 0.001). Furthermore, several
taxon-specific RAPD markers
were identified for both species. In contrast, no
specific DNA fragments were detected that could
link genetic markers to the geographical origins of
the sampled populations. These findings provide
molecular evidence supporting the recognition of
H. medicinalis and H. verbana as distinct species
[14-15]
.
In molecular systematics studies by Utevsky and
Trontelj, they investigated phylogenetic
relationships among several Hirudo species,
including specimens collected in Azerbaijan. The
analyses employed both nuclear markers (ITS2 and
5.8S rRNA) and mitochondrial markers (12S rRNA
and COI). The results supported the monophyly of
the genus Hirudo, indicating that its constituent
species share a common evolutionary ancestry
[18]
(Figure1.1.).
Figure 1.1. Phylogenetic tree of medicinal
leeches.
We constructed the phylogenetic tree based on
molecular analyses of species in the genus Hirudo,
using mitochondrial markers including the
cytochrome c oxidase subunit I (COI) gene and 12S
rRNA. The results indicate that, despite their
morphological similarities, H. medicinalis, H.
verbana, and H. orientalis represent genetically
distinct species. In contrast, H. nipponia and H.
troctina exhibit more distant phylogenetic
relationships relative to these three species. The
clustering of specimens collected from different
geographical regions within the same phylogenetic
clade suggests shared evolutionary ancestry,
although additional evidence is required to establish
the role of geographical isolation in their
divergence. The phylogenetic position of H.
orientalis is consistent with its association with the
Issue : 1
INTERNATIONAL JOURNAL OF DIAGNOSTICS AND RESEARCH [ISSN No.: 2584-2757]
Volume : 4
Copyright @ : - Dr.Laman Dadashova Inter. J.Digno. and Research IJDRMSID0144 |ISSN :2584-2757
53
Caspian region. Overall, molecular marker-based
systematics provides an essential framework for
resolving taxonomic relationships and improving
the accuracy of species identification within the
genus Hirudo. A 2020 study investigated the
genome of Hirudo medicinalis and examined gene
expression in salivary cells isolated from H.
medicinalis, H. orientalis, and H. verbana using
RNA sequencing (RNA-seq) and proteomic
analyses. The study identified numerous salivary-
cell-associated genes, including those encoding
previously uncharacterized salivary components.
Furthermore, the findings revealed that genes
encoding known anticoagulant proteins are
expressed not only in salivary cells but also in other
cell types. These results provide important insights
into the molecular and genetic mechanisms
underlying blood feeding in medicinal leeches and
contribute to a better understanding of the roles of
salivary proteins in regulating the host blood
coagulation system.
In a study conducted by Sh. Farzali and N. Saglam,
the mitochondrial cytochrome c oxidase subunit I
(COI) gene region was analysed in medicinal
leeches collected from the Lankaran–Astara region
of Azerbaijan. The results revealed 100% sequence
identity between the analyzed DNA sequences and
those of Hirudo orientalis Utevsky & Trontelj,
2005, confirming the molecular identification of the
specimens as this species
[4]
. A separate study
demonstrated that microsatellite profiles differ
between Hirudo medicinalis and Hirudo verbana,
with species-specific differentiation detected at four
microsatellite loci. Two loci, Hm8 and Hm12,
failed to amplify in H. verbana, whereas HvA10
and HvH07 failed to amplify in H. medicinalis.
Among the other European medicinal leech species
examined, three of the five successfully amplified
loci—Hm2, Hm10, and HvT397—were
monomorphic in H. medicinalis but polymorphic in
H. verbana. DNA barcoding further supported
these findings, identifying distinct monophyletic
groups for the two species and indicating that they
represent separate evolutionary lineages, with an
average sequence divergence of 8.5% between
them
[13]
.
A study conducted in Iraq examined 19 individuals
of the genus Hirudo. We amplified the ITS-2,
mitochondrial cytochrome c oxidase subunit I
(COI), and mitochondrial 12S ribosomal RNA (12S
rRNA) markers using polymerase chain reaction
(PCR), then sequenced the resulting products.
Comparison of ITS-2
sequences revealed a single genotype in both
Hirudo verbana and Hirudo orientalis samples. In
contrast, COI sequence analysis identified four
haplotypes in H. verbana and two in H. orientalis.
Analysis of the 12S rRNA marker revealed two
haplotypes in H. verbana, whereas only one
haplotype was detected in H. orientalis. Overall,
the study provides additional insights into the
genetic variation and differentiation of Hirudo
species used in medical clinics in Iraq.
Conclusion :
Findings from molecular and phylogenetic studies
indicate that Hirudo orientalis represents a distinct
species, genetically differentiated from other
medicinal leeches despite their morphological
similarities. Evidence concerning its therapeutic
potential, capacity to utilize diverse food sources,
Issue : 1
INTERNATIONAL JOURNAL OF DIAGNOSTICS AND RESEARCH [ISSN No.: 2584-2757]
Volume : 4
Copyright @ : - Dr.Laman Dadashova Inter. J.Digno. and Research IJDRMSID0144 |ISSN :2584-2757
54
and transcriptomic variation further highlights its
potential pharmacological and medical
significance. These findings underscore the
importance of integrating molecular-genetic
markers into the taxonomy of Hirudo, rather than
relying exclusively on morphological
characteristics. This integrative approach enables
more robust taxonomic identification, supports
accurate assessment of the conservation status of
natural populations, and provides a scientific basis
for developing standardised laboratory breeding
protocols and improving the responsible use of
medicinal leeches in clinical practice.
References:
1. Darabi-Darestani, K., & Malek, M. (2011).
Seasonal variation in the occurrence of the
medicinal leech Hirudo orientalis in Guilan
Province, Iran. Aquatic Biology, 11(3), 289–
294.
2. Darabi-Darestani, K., Sari, A., Sarafrazi, A.,
et al. (2018). Entrapped by the uneven
central and Middle Eastern terrains: Genetic
status of populations of Hirudo orientalis
(Annelida, Clitellata, Hirudinida) with a
phylogenetic review of the genus Hirudo.
Molecular Phylogenetics and Evolution,
121, 52–60.
3. Elliott, J. M., & Kutschera, U. (2011).
Medicinal leeches: Historical use, ecology,
genetics and conservation. Freshwater
Reviews, 4(1), 21–41.
4. Farzali, Sh., & Saglam, N. (2020). The
status of the leech fauna (Annelida,
Hirudinea) in the eastern region of
Azerbaijan. Journal of Wildlife and
Biodiversity, 4(4), 40–52.
5. Fedorova, L., & Kaygorodova, I. (2016).
The present state of the leech fauna
(Annelida, Hirudinea) in the Upper Irtysh
cascade of water reservoirs. ZooKeys, 596,
1–12.
6. Habeeb, K. B., & Al-Warid, H. S. (2024).
Molecular study of two medicinal leech
species. Research Journal of Chemistry and
Environment, 28(3), 1363–1378.
7. Hildebrandt, J. P., & Lemke, S. (2011).
Small bite, large impact—saliva and
salivary molecules in the medicinal leech,
Hirudo medicinalis. Naturwissenschaften,
98(12), 995–1008.
8. Lai, Y. T., Nakano, T., & Chen, J. (2011).
Three species of land leeches from Taiwan,
Haemadipsa rjukjuana comb. n., a new
record for Haemadipsa picta Moore, and an
updated description of Tritetrabdella
taiwana (Oka). Ecological Research, 26, 1–
22.
9. Liu, Y. F., Wang, W. D., Xie, T., Chen, L.
M., Zhang, Q. G., Lou, W. J., Wang, Z. M.,
Ma, Y., & Gao, H. M. (2026). A
multidimensional strategy for exploring
Issue : 1
INTERNATIONAL JOURNAL OF DIAGNOSTICS AND RESEARCH [ISSN No.: 2584-2757]
Volume : 4
Copyright @ : - Dr.Laman Dadashova Inter. J.Digno. and Research IJDRMSID0144 |ISSN :2584-2757
55
active peptides combining chemical analysis
and bioassays for medicinal leeches.
Journal of Ethnopharmacology, 371, Article
122113.https://doi.org/10.1016/j.jep.2026.1
22113
10. Mayr, E. (1963). Animal species and
evolution. Harvard University Press.
11. PetrauskienÄ—, L. (2004). The medicinal
leech as a convenient tool for water toxicity
assessment. Environmental Toxicology,
19(4), 336–341.
12. PetrauskienÄ—, L., Utevska, O., & Utevsky,
S. (2009). Can different species of
medicinal leeches (Hirudo spp.) interbreed?
Invertebrate Biology, 128(4), 324–331.
13. Siddall, M. E., & Kuge, A. G. (1997).
Probabilism and phylogenetic inference.
Cladistics, 13(4), 313–336.
14. Siddall, M. E., Borda, E., & Budinoff, R. B.
(2005). Phylogenetic evaluation of
systematics and biogeography of the leech
family Glossiphoniidae. Invertebrate
Systematics, 19(2), 105–112.
15. Siddall, M. E., Trontelj, P., Utevsky, S. Y.,
et al. (2007). Diverse molecular data
demonstrate that commercially available
medicinal leeches are not Hirudo
medicinalis. Proceedings of the Royal
Society B: Biological Sciences, 274(1617),
1481–1487.
16. The IUCN Red List of Threatened Species.
(2026). Hirudo medicinalis Linnaeus, 1758
(Version 2026-1).
https://www.iucnredlist.org
17. Trontelj, P. (2012). Phylogeny and
phylogeography of medicinal leeches
(genus Hirudo): Fast dispersal and shallow
genetic structure. Molecular Phylogenetics
and Evolution, 63(2), 475–485.
18. Utevsky, S. Y., Kovalenko, N., &
Doroshenko, K. (2009). Chromosome
numbers for three species of medicinal
leeches (Hirudo spp.). Systematic
Parasitology, 74(2), 95–102.
19. Utevsky, S. Y., Zagmajster, M., &
Atemasov, A. (2010). Distribution and
status of medicinal leeches (genus Hirudo)
in the Western Palaearctic: Anthropogenic,
ecological, or historical effects? Aquatic
Conservation: Marine and Freshwater
Ecosystems, 20(2), 198–210.
Declaration :
Conflict of Interest : None
Issue : 1
INTERNATIONAL JOURNAL OF DIAGNOSTICS AND RESEARCH [ISSN No.: 2584-2757]
Volume : 4
Copyright @ : - Dr.Laman Dadashova Inter. J.Digno. and Research IJDRMSID0144 |ISSN :2584-2757
56
ISSN: 2584-2757
DOI : :10.5281/zenodo.23434757
Dr.Laman Dadashova Inter. J.Digno. and Research
This work is licensed under Creative
Commons Attribution 4.0 License
Submission Link : http://www.ijdrindia.com
Benefits of Publishing with us
Fast peer review process
Global archiving of the articles
Unrestricted open online access
Author retains copyright
Unique DOI for all articles
https://ijdrindia.com