Citridiol is an extract of the leaves of Corymbia citriodora (Myrtaceae), the lemon eucalyptus, and mostly consists of p-menthane-3,8-diol isomers. The effectiveness of this extract as a repellent against land leeches of the genus Haemadipsa (Haemadipsidae), primarily H. sylvestris, was tested in the laboratory and field, in Peninsular Malaysia. The formulation tested, Mosi-guard Natural spray, contained 40% (w/w) Citridiol in a base of ethanol, water and isopropanol. In the laboratory test, specimens of H. sylvestris that were placed within moist, untreated arenas enclosed by treated paper rings made numerous attempts to cross the rings but were prevented or delayed from crossing over, in a dose-dependent manner. Mortality was high among the leeches that attempted to cross over the paper rings that had been sprayed to saturation point but low among the leeches that attempted to cross over paper rings that had only been partially treated, with a droplet-spray. The field study was carried out using indices that were formulated to reflect the severity of leech attack and the degree of repellency. Heavy or moderate spraying of footwear and trouser legs (tucked into socks) not only gave complete protection against bites by H. sylvestris and H. picta but also provided high enough repellency to keep the treated footwear virtually free of leeches. Even a light spray greatly reduced the numbers of leeches on footwear and delayed their progression toward biting the test subjects, although it failed to prevent bites completely. There was no decline in the repellency of the Citridiol when hourly assessments were made over a 6-h test period in the field. The results of the study show that Citridiol is highly repellent as well as toxic to leeches, and can be effectively used to prevent leech bites in the field.
In this study the genetic diversity of local freshwater leeches (Hirudinaria spp.) was inferred using mtDNA COI gene analysis and compared with the gross external variations of 26 freshwater leech specimens obtained from the wild and leech farms. Based on a neighbor-joining tree generated from 516 COI base sequences, four distinct clades of Hirudinaria were seen with interspecific genetic divergence in the range of 7.6-14.5%. The external morphological variations based on the presence of stripes, location of gonopores, and anus separated the samples into four morphologically distinct groups matching the four clades obtained from the molecular data. Two black stripes at the ventral region were observed only in specimens found clustered with clades that contained the GenBank-reported H. manillensis, whereas the brown or dark green coloration without stripes on the ventral region was seen in samples that clustered with H. javanica and H. bpling clades.
The anticoagulant effect of leech saliva was traditionally employed in the treatment of diabetes mellitus complications such as peripheral vascular complications. This study was carried out to examine the effect of leech saliva extract (LSE) on blood glucose levels in alloxan-induced diabetic rats. First, LSE was collected from leeches which were fed on a phagostimulatory solution. Second, total protein concentration was estimated using the Bradford assay. Third, diabetic rats were injected subcutaneously (sc) with LSE at doses of 500 and 1 000 μg·kg(-1) body weight (bw). Other diabetic rats were injected sc with insulin at doses of 10 and 20 U·kg(-1) bw. Another group was injected simultaneously with LSE (250 μg·kg(-1) bw) and insulin (10 U·kg(-1) bw). Fasting blood glucose (FBG) concentrations were monitored during a study period of eight hours at regular intervals. Findings showed that both doses of LSE resulted in a significant and gradual decrease in FBG starting from 10%-18% downfall after two hours of injection reaching the maximal reduction activity of 58% after eight hours. Remarkably, LSE was sufficient to bring the rats to a near norm-glycemic state. The high dose of insulin induced a severe hypoglycemic condition after 2-4 h of injection. The lower dose was able to decline FBG for 2-6 h in rats which became diabetic again after 8 h. On the other hand, the concurrent injection of low doses of LSE and insulin produced a hypoglycemic effect with all rats showing normal FBG levels. Taken together, these findings indicated that the subcutaneous injection of LSE of the medicinal Malaysian leech was able to provide better glycemic control compared with insulin. Moreover, the synergism between LSE and insulin suggests that LSE could be utilized as an adjuvant medication in order to reduce insulin dosage or to achieve better control of blood glucose.
Haemadipsid leeches are among the most successful terrestrial invertebrates in Bornean rainforests. They are very common ectoparasites of vertebrates, and their abundance has facilitated the conduction of numerous projects in the fields of ecology, zoogeography and taxonomy. We undertook research on two species inhabiting lowland dipterocarp forest, Haemadipsa picta Moore, 1929 and Haemadipsa subagilis (Moore, 1929), in order to address the following questions: (a) is there a difference in leech abundance between trails and off-trails?; (b) is ambush location dependent on specimen size or is species-specific?; (c) is intra- and interspecific competition limited by differences in foraging behaviours or vertical niche partitioning? Our results clearly show that H. picta is more abundant on trails than on off-trails and is vertically dispersed within the understory; the size of a specimen is strongly correlated with plant height. Haemadipsa subagilis was found not to exhibit such patterns. We suggest a possible lowering of interspecific competition between these species as a result of: (i) size-dependent dispersion of H. picta (together with reduction of intraspecific competition); and (ii) habitat specialisation of H. subagilis. Moreover, we provide new observations on their foraging behaviour.
Hematophagous animals including leeches have been known to possess biologically active compounds in their secretions, especially in their saliva. The blood-sucking annelids, leeches have been used for therapeutic purposes since the beginning of civilization. Ancient Egyptian, Indian, Greek and Arab physicians used leeches for a wide range of diseases starting from the conventional use for bleeding to systemic ailments, such as skin diseases, nervous system abnormalities, urinary and reproductive system problems, inflammation, and dental problems. Recently, extensive researches on leech saliva unveiled the presence of a variety of bioactive peptides and proteins involving antithrombin (hirudin, bufrudin), antiplatelet (calin, saratin), factor Xa inhibitors (lefaxin), antibacterial (theromacin, theromyzin) and others. Consequently, leech has made a comeback as a new remedy for many chronic and life-threatening abnormalities, such as cardiovascular problems, cancer, metastasis, and infectious diseases. In the 20(th) century, leech therapy has established itself in plastic and microsurgery as a protective tool against venous congestion and served to salvage the replanted digits and flaps. Many clinics for plastic surgery all over the world started to use leeches for cosmetic purposes. Despite the efficacious properties of leech therapy, the safety, and complications of leeching are still controversial.
The high prevalence (80-100%) of the marine leech Zeylanicobdella arugamensis (De Silva) on cage-cultured Asian sea bass Lates calcarifer (Bloch) led us to investigate the percentage of juvenile leeches hatched from deposited cocoons, survival of juvenile and adult marine leeches at different salinity and temperature. The results showed that the hatching percentage of juvenile leeches was highest at salinity of 30 ppt (32.5 ± 2.8%) followed by 20 ppt (18.0 ± 4.3%) and 10 ppt (12.1 ± 1.4%), respectively. It was found that the adult and juvenile leeches could live up to an average range of 4-7 days at salinity ranging from 10 to 40 ppt. The juvenile leeches were able to hatch at temperature ranging from 25 to 35 °C but unable to hatch at 40 °C. The survival period of adult and juvenile leeches ranged from 11 to 16 days at 25 °C, which was comparatively longer than 5-13 days and 10 h--5 days at 27-30 °C and 35-40 °C, respectively. The study provided the information on the physical parameters of salinity and temperature which are most optimal for the marine leech Z. arugamensis to propagate.
Leech infestation in otolaryngology is sporadically seen in
remote area of tropical countries, but the cases are usually not
scientifically reported. We are reporting an unusual case of a
land leech removal from external auditory canal (EAC).
Cellulitis in human following leech bite is rae, many of the case reports were associated with the use of medical leech. We report the first documented case of cellulitis following the bite of land leech (Haemodipsa species), known locally as 'pacat', in a 2 month-old-baby. The baby developed fever but subsided following an antibiotic cover. The baby had uneventful stay in the hospital and the cellulitis was successfully treated following a week cource of Cloxacillin.
Leech is an uncommon nasal foreign body, which can cause epistaxis. It is an aquatic segmental
worm living in fresh water most commonly in tropical areas. Once it is attached in the nose, it will
secrete an anticoagulant enzyme named hirudin, and that will result in continuous bleeding more
than from a normal wound even after it is removed. We present a case of a live leech in the nose
that caused unilateral epistaxis in a patient with a recent history of jungle trekking.
Leech is an uncommon nasal foreign body, which can cause epistaxis. It is an aquatic segmental worm living in fresh water most commonly in tropical areas. Once it is attached in the nose, it will secrete an anticoagulant enzyme named hirudin, and that will result in continuous bleeding more than from a normal wound even after it is removed. We present a case of a live leech in the nose that caused unilateral epistaxis in a patient with a recent history of jungle trekking.
The application of metabarcoding to environmental and invertebrate-derived DNA (eDNA and iDNA) is a new and increasingly applied method for monitoring biodiversity across a diverse range of habitats. This approach is particularly promising for sampling in the biodiverse humid tropics, where rapid land-use change for agriculture means there is a growing need to understand the conservation value of the remaining mosaic and degraded landscapes. Here we use iDNA from blood-feeding leeches (Haemadipsa picta) to assess differences in mammalian diversity across a gradient of forest degradation in Sabah, Malaysian Borneo. We screened 557 individual leeches for mammal DNA by targeting fragments of the 16S rRNA gene and detected 14 mammalian genera. We recorded lower mammal diversity in the most heavily degraded forest compared to higher quality twice logged forest. Although the accumulation curves of diversity estimates were comparable across these habitat types, diversity was higher in twice logged forest, with more taxa of conservation concern. In addition, our analysis revealed differences between the community recorded in the heavily logged forest and that of the twice logged forest. By revealing differences in mammal diversity across a human-modified tropical landscape, our study demonstrates the value of iDNA as a noninvasive biomonitoring approach in conservation assessments.
The application of high-throughput sequencing (HTS) for metabarcoding of mixed samples offers new opportunities in conservation biology. Recently, the successful detection of prey DNA from the guts of leeches has raised the possibility that these, and other blood-feeding invertebrates, might serve as useful samplers of mammals. Yet little is known about whether sympatric leech species differ in their feeding preferences, and whether this has a bearing on their relative suitability for monitoring local mammalian diversity. To address these questions, we collected spatially matched samples of two congeneric leech species Haemadipsa picta and Haemadipsa sumatrana from lowland rainforest in Borneo. For each species, we pooled ~500 leeches into batches of 10 individuals, performed PCR to target a section of the mammalian 16S rRNA locus and undertook sequencing of amplicon libraries using an Illumina MiSeq. In total, we identified sequences from 14 mammalian genera, spanning nine families and five orders. We found greater numbers of detections, and higher diversity of OTUs, in H. picta compared with H. sumatrana, with rodents only present in the former leech species. However, comparison of samples from across the landscape revealed no significant difference in mammal community composition between the leech species. We therefore suggest that H. picta is the more suitable iDNA sampler in this degraded Bornean forest. We conclude that the choice of invertebrate sampler can influence the detectability of different mammal groups and that this should be accounted for when designing iDNA studies.
Zeylanicobdella arugamensis (Hirudinea), a marine parasitic leech, not only resulted in the mortality of the host fish (Groupers) but also caused economic losses. The current study aimed to elucidate the antiparasitic efficacy of the aqueous extract of the Azadirachta indica leaves against Z. arugamensis and to profile the composition via LC-Q Exactive HF Orbitrap mass spectrometry. Different concentrations (25, 50 and 100 mg/mL) of A. indica extract were prepared and tested on the parasitic leeches. The total mortality of leeches was noticed with an exposure to the A. indica aqueous extract. The average times required for the aqueous extract at concentrations of 25, 50 and 100 mg/mL to kill the leeches were 42.65 ± 9.20, 11.69 ± 1.11 and 6.45 ± 0.45 min, respectively, in a dose-dependent manner. The Orbitrap mass spectrometry analysis indicated the presence of five flavonoids (myricetin 3-O-galactoside, trifolin, isorhamnetin, quercetin and kaempferol), four aromatics (4-methoxy benzaldehyde, scopoletin, indole-3-acrylic acid and 2,4-quinolinediol), three phenolics (p-coumaric acid, ferulic acid and phloretin) and two terpenoids (pulegone and caryophyllene oxide). Thus, our study indicates that A. indica aqueous extract is a good source of metabolites with the potential to act as a biocontrol agent against the marine parasitic leech in aquaculture.
The medicinal Malaysian leeches have been used in traditional medicine to treat many different ailments. In this study, leech saliva extract (LSE) was collected from the medicinal Malaysian leech Hirudinaria manillensis. Gel electrophoresis of LSE was carried out to estimate the peptide and protein molecular weights of its content. Results showed that LSE contains more than 60 peptides and proteins with molecular masses ranging from 1.9-250kDa. Thrombin time assay in vitro was employed to assess the collected LSE antithrombin activity. First, to study its stability, LSE was lyophilized under the following different conditions: pre-freezing temperature, type of container and lyophilization cycle. Pre-freezed LSE sample at -20°C and lyophilized for 24 hours retained about 100-95% of its original biological activities. Second, the LSE antithrombin activity was monitored for a period of six months. Storage temperature, type of the container and photosensitivity effects on antithrombin activity of the lyophilized (solid state) and non-lyophilized (liquid state) were investigated. Results showed that storage temperature drastically affected the biological activity of LSE with -20 °C as the optimum temperature. Samples stored at ambient temperature and +4 °C were light photosensitive and adversely affected when stored in polypropylene tubes. Lyophilized samples were more stable than non-lyophilized ones over the period of study. To sum up, in order to have a biologically active stock of LSE, it has to be lyophilized for no more than 24 hours following freezing at -20°C and has to be stored at -20°C in glass tubes protected from light.
Marine leech Zeylanicobdella arugamensis (Piscicolidae), an economically important parasite is infesting predominantly cultured groupers, hybrid groupers and other fish in Southeast Asian countries. In this study, we tested the anti-parasitic potential of a medicinal plant Nephrolepis biserrata found in Sabah, East Malaysia against Z. arugamensis. Various concentrations of methanol extracts of the plant were tested experimentally against Z. arugamensis and disinfestation of the leech from its primary host hybrid groupers. The composition of methanol extract of N. biserrata was determined through LC-QTOF analysis. The significant anti-parasitic activity of 100% mortality of leeches was observed with the exposure of N. biserrata extracts. The average time to kill the leeches at concentrations of 25, 50 and 100 mg/ml was 25.11 ± 3.26, 11.91 ± 0.99, and 4.88 ± 0.50 min., respectively. Further, at various low concentrations of N. biserrata 2.5, 5 and 10 mg/ml, hybrid groupers were disinfested in an average time of 108.33 ± 12.65, 65.83 ± 9.70 and 29.16 ± 5.85 min., respectively. The tandem mass spectrometry data from LC-QTOF indicated some hits on useful bioactive compounds such as terpenoids (ivalin, isovelleral, brassinolide, and eschscholtzxanthin), flavonoids (alnustin, kaempferol 7,4'-dimethyl ether, and pachypodol), phenolics (piscidic acid, chlorogenic acid, and ankorine), and aromatic (3-hydroxycoumarin). Thus N. biserrata can act as a potential biocontrol agent.
A 3-year old girl presented to our A&E with a complaint of an acute event of per rectal bleed. The parent had given a history of a visit to a nearby waterfall one day ago. Upon admission she developed hypotension and had a fainting episode due to the excessive passing of blood per rectum. The case report shows how the appropriate investigations were carried out and the twist behind the cause.