Horseshoe crabs are among the most studied invertebrates due to their unique, innate immune system and biological processes. The metabolomics study was conducted on lipopolysaccharide (LPS)-stimulated and non-stimulated hemocytes isolated from the Malaysian Tachypleus gigas and Carcinoscorpius rotundicauda. LC-TOF-MS, multivariate analyses, principal component analysis (PCA), and partial least squares-discriminant analysis (PLS-DA) were included in this study to profile the metabolites. A total of 37 metabolites were identified to be differentially abundant and were selected based on VIP > 1. However, of the 37 putative metabolites, only 23 were found to be significant with ANOVA at p
Despite being widely distributed in Asia, Carcinoscorpius rotundicauda is often overlooked and, its population status remains unclear. Moreover, it is threatened by illegal harvesting and degradation of mangrove ecosystems. Protecting its habitat is essential for population and biodiversity conservation, as mangroves provide nursery grounds and food supply for C. rotundicauda. This review discusses the biological characteristics of C. rotundicauda, including ecology, nutrition, life history, toxicology, and immunology. It also presents information about its distribution and population status. The review emphasizes the challenges faced by C. rotundicauda and proposes a conservation framework that involves the participation of local residents to facilitate conservation efforts. Collaboration between local residents and communities is proposed to protect and monitor the mangrove ecosystem. Additionally, this framework can support field research, protect C. rotundicauda juveniles and other species, and ensure the livelihood of local residents through participation in carbon trading markets and eco-industries such as eco-farming and eco-tourism.
It has been widely reported that allozyme frequency variation is a potential indicator of heavy metal-induced impacts in aquatic populations. In the present study, wild populations of horseshoe crab (Carcinoscorpius rotundicauda) were collected from contaminated and uncontaminated sites of Peninsular Malaysia. By adopting horizontal starch gel electrophoresis, seven enzyme systems were used to study allozyme polymorphisms. Nine polymorphic loci were observed in C. rotundicauda. The relationships of allozyme variations with the concentrations of Cd, Cu, Ni, and Zn in sediments and in muscle tissues of horseshoe crabs were determined. Based on genetic distance, the lower mean value of Nei's D (0.017) indicated that both of the contaminated populations of Kg. Pasir Puteh and Kuala Juru were very closely related when compared to the relatively uncontaminated Pantai Lido population. Higher heterozygosities were shown by the contaminated populations when compared to the uncontaminated population. Different allelic frequencies could be observed for the aldolase (ALD; E.C. 2.7.5.1) locus between the contaminated and uncontaminated populations of C. rotundicauda. The dendrogram of genetic relationships of the three populations of C. rotundicauda showed the same clustering pattern as the dendrograms are based on heavy metals in the sediments and in the horseshoe crabs' abdominal muscles. From the F statistics, the present study showed that the three populations of horseshoe crabs were considered to have undergone moderate genetic differentiation with a mean F (ST) value of 0.092 .The current results suggest that allozyme polymorphism in horseshoe crabs is a potential biomonitoring tool for metal contamination, although further validation is required.
Horseshoe crabs (HSCs) are living fossil species of marine arthropods with a long evolutionary history spanning approximately 500 million years. Their survival is helped by their innate immune system that comprises cellular and humoral immune components to protect them against invading pathogens. To help understand the genetic mechanisms involved, the present study utilised the Illumina HiSeq platform to perform transcriptomic analysis of hemocytes from the HSC, Tachypleus gigas, that were challenged with lipopolysaccharides (LPS). The high-throughput sequencing resulted in 352,077,208 and 386,749,136 raw reads corresponding to 282,490,910 and 305,709,830 high-quality mappable reads for the control and LPS-treated hemocyte samples, respectively. Based on the log-fold change of > 0.3 or
Local and regional decline of Asian horseshoe crabs has spurred a study on its spawning population at Balok Beach, Kuantan, Pahang, Malaysia. This location was identified as spawning site due to the occurrence of horseshoe crab spawning pairs and nests. Size-frequency, length-weight relationships, sex ratio and epibiont infestation of Tachypleus gigas were studied. Instar stage was estimated based on prosomal width. Condition of the horseshoe crab carapace was reported. Visual search technique of horseshoe crab was conducted during high tide of new and full moons. Prosomal, opisthosomal and telson length and weight of each horseshoe crab were measured. Largest female was recorded with mean prosomal length and width of 154.4 and 246.9 mm, respectively. About 69.8% of the males belonged to size group of 151-200 mm and 53.3% of females were grouped into 201-250 mm. All individuals were of fourteenth to sixteenth instar stages. Sex ratio varied from 0.313 to 2.5 and attributed to commercial harvest and monsoon season. Sand sediment of study site showed 93% of fine sands with grain size ranged from 120 to 250 microm. Acorn and pedunculate barnacle, conical and flat slipper shells were found on the carapace of the specimens. Most males had damaged eyes and carapaces while females with broken telsons. Body damages of about 19.9% on the specimens were likely due to nearby fishing activities. Lack of satellite male indicated low spawning population. The finding of this study showed that the species is extremely threatened by human activities and coastal development.
The horseshoe crab (Carcinoscorpius rotundicauda) is consumed by those residing near the coastal areas of Kota Marudu District in Malaysia, as it is considered a delicacy. During June to August, 2011 thirty cases of tetrodotoxin poisoning were reported from Kota Marudu District following ingestion of horseshoe crabs caught in Kota Marudu Bay. The purpose of this study is to describe this case series in order to determine risk factors to prevent further outbreaks. There were six confirmed and 24 probable cases of tetrodotoxin poisoning identified in the study area during the study period as diagnosed by clinical presentation and laboratory findings. Symptoms included dizziness (80%), circumoral and lingual numbness (80%), hand and feet numbness (63.3%), nausea and vomiting (30%) and weakness and difficulty in breathing (26.6%). Three cases (10%) died while 27 cases recovered. Forty-seven percent of the cases had onset of symptoms within 30 minutes of ingestion and 14% 31-60 minutes after ingestion of horseshoe crab meat. Urine samples were collected from the cases, while horseshoe crabs, cockles and sea water from the epidemic area were also taken for analysis. Tetrodotoxin was detected in the urine of six cases; the highest concentrations recorded were among the three cases who died. High tetrodotoxin concentrations were found in the hepatic cecum and eggs of the tested horseshoe crabs. Dinoflagellates were not detected in the sea water or cockle samples. Intensive health education was initiated quickly to stop other members of the Marudu Bay community from consuming the horseshoe crabs. This is the first documented epidemic of tetrodotoxin poisoning in Sabah.
Whole genome duplication (WGD) has occurred in relatively few sexually reproducing invertebrates. Consequently, the WGD that occurred in the common ancestor of horseshoe crabs ~135 million years ago provides a rare opportunity to decipher the evolutionary consequences of a duplicated invertebrate genome. Here, we present a high-quality genome assembly for the mangrove horseshoe crab Carcinoscorpius rotundicauda (1.7 Gb, N50 = 90.2 Mb, with 89.8% sequences anchored to 16 pseudomolecules, 2n = 32), and a resequenced genome of the tri-spine horseshoe crab Tachypleus tridentatus (1.7 Gb, N50 = 109.7 Mb). Analyses of gene families, microRNAs, and synteny show that horseshoe crabs have undergone three rounds (3R) of WGD. Comparison of C. rotundicauda and T. tridentatus genomes from populations from several geographic locations further elucidates the diverse fates of both coding and noncoding genes. Together, the present study represents a cornerstone for improving our understanding of invertebrate WGD events on the evolutionary fates of genes and microRNAs, at both the individual and population level. We also provide improved genomic resources for horseshoe crabs, of applied value for breeding programs and conservation of this fascinating and unusual invertebrate lineage.
An outbreak of the sessile peritrich Zoothamnium duplicatum in a pilot, commercial-scale Limulus polyphemus hatchery resulted in the loss of ∼96% (40,000) second/third instar larvae over a 61day period. peritrich growth was heavy, leading to mechanical obstruction of the gills and physical damage. The peritrichs were controlled without resultant loss of juvenile crabs by administering 10ppm chlorine in freshwater for 1h and the addition of aquarium grade sand; a medium into which the crabs could burrow and facilitate cleaning of the carapace. Peritrich identity was confirmed from a partial SSU rDNA contiguous sequence of 1343bp (99.7% similarity to Z. duplicatum).
The central coast of Tamil Nadu, Andaman and Nicobar Islands, along with other regions, was severely hit by the tsunami on 26 December 2004. A strange spawning behaviour of the Indian horseshoe crabs (Tachypleus gigas, Müller) was observed after the tsunami disturbance. Along the northeast coast of India, the horseshoe crabs normally migrate towards the shore in large numbers coinciding with the tidal height and grain size of the sediment and spawn in nests made in sand. The spawning behaviour was normal until November 2004 and throughout that period about 35 to 45 nests in 200 m2 of area of the breeding beach were examined. However, no nesting was observed from December 2004 to March 2005, i.e. after the occurrence of tsunami. The number of nests per 200 m2 of area (~82) and the number of eggs per nest (750-1000 eggs) were increased considerably when the normal conditions were restored in April 2005. The absence of spawning migration, from December 2004 to March 2005, is totally an unusual behaviour and this happened for the first time during the past several years of the study period.
Known as ‘belangkas’ in Malaysia, horseshoe crabs have been used by locals for food, bait and as fertilizer. Currently, these ancient mariners are gaining a lot of attention as the amoebocyte from their blood are harvested by the biomedical industry for use to detect human pathogens in patients, injectable drugs and intravenous devices.
Endotoxins (chemically known as Lipopolysaccharide) from gram-negative microorganisms initiates clot formation in blood when it is accidentally encountered by horseshoe crab blood stream. This property was extensively studied by various researchers as a result Limulus Amebocyte Lysate (LAL) test was established. The LAL tests in general, 3 to 300 times more sensitive than the United States Pharmacopeial (USP) rabbit pyrogen test method. It is apparent that major differences among the LAL preparations lie in the area of sensitivity. Differences, up to 100-fold, exist in the sensitivity of the various LAL preparations to the same endotoxin. Based on the above perspective, a portable Kit (Endo sensor) was developed to detect the presence of bacterial endotoxin in liquid biological samples using Tachypleus Amebocyte Lysate (TAL) as a source. Sensitivity of the Kit was determined using various concentrations of prepared endotoxin standards and pyrogen free water samples. It was observed that Endo sensor could detect up to nano gram level of endotoxin in liquid biological samples which could be expressed in (EU/ml) and the labeled sensitivity of the lysated product was 0.125 EU/ml. The gel clotting principle method was utilized for the detection of bacterial endotoxin in liquid biological samples.
Tanjung Selongor and Pantai Balok (State Pahang) are the only two places known for spawning activity of the Malaysian horseshoe crab - Tachypleus gigas (Müller, 1785) on the east coast of Peninsular Malaysia. While the former beach has been disturbed by several anthropogenic activities that ultimately brought an end to the spawning activity of T. gigas, the status of the latter remains uncertain. In the present study, the spawning behavior of T. gigas at Pantai Balok (Sites I-III) was observed over a period of thirty six months, in three phases, between 2009 and 2013. Every year, the crab's nesting activity was found to be high during Southwest monsoon (May-September) followed by Northeast (November-March) and Inter monsoon (April and October) periods. In the meantime, the number of female T. gigas in 2009-2010 (Phase-1) was higher (38 crabs) than in 2010-2011 (Phase-2: 7 crabs) and 2012-2013 (Phase-3: 9 crabs) for which both increased overexploitation (for edible and fishmeal preparations) as well as anthropogenic disturbances in the vicinity (sand mining since 2009, land reclamation for wave breaker/parking lot constructions in 2011 and fishing jetty construction in 2013) are responsible. In this context, the physical infrastructure developments have altered the sediment close to nesting sites to be dominated by fine sand (2.5Xφ ) with moderately-well sorted (0.6-0.7σφ), very-coarse skewed (-2.4SKφ), and extremely leptokurtic (12.6Kφ) properties. Also, increased concentrations of Cadmium (from 4.2 to 13.6 mg kg(-1)) and Selenium (from 11.5 to 23.3 mg kg(-1)) in the sediment, and Sulphide (from 21 to 28 µg l(-1)) in the water were observed. In relation to the monsoonal changes affecting sheltered beach topography and sediment flux, the spawning crabs have shown a seasonal nest shifting behaviour in-between Sites I-III during 2009-2011. However, in 2012-2013, the crabs were mostly restricted to the areas (i.e., Sites I and II) with high oxygen (5.5-8.0 mg l(-1)) and moisture depth (6.2-10.2 cm). In view of the sustained anthropogenic pressure on the coastal habitats on one hand and decreasing horseshoe crabs population on the other, it is crucial to implement both conservation and management measures for T. gigas at Pantai Balok. Failing that may lead to the loss of this final spawning ground on the east coast of P. Malaysia.
Respiratory metabolism of the larvae of Malaysian horseshoe crab Tachypleus gigas (Müller) was studied under different salinities, pH, and temperature. The trend in oxygen consumption was uniform at all salinities, ranging from 10-40 ppt, indicating insignificant influence on the oxygen consumption by the larvae. Similarly, the correlation coefficient values showed that the relationship between oxygen consumption and salinity was not significant (P > 0.05; r = 0.245). During the first three hours, the oxygen consumption was 8.89, 10.72, 17.4, and 12.06% at 10, 20, 30, and 40 ppt salinities, respectively. Meanwhile, the maximum oxygen consumption was recorded after 12 hrs, i.e. at salinity 20 ppt. A sudden drop in oxygen consumption was recorded during 3-6 hours of the experiment. This was followed by a gradual increase in the consumption of oxygen up to 12 hours of experiment. A similar trend in the oxygen consumption was observed in different pH levels, ranging from 5 to 9. At pH 6 and 9, during the first six hour, a moderate consumption of oxygen was observed. However, at pH 6, 7 and 8, the rates of oxygen consumption were found to be relatively greater after six hours, indicating unfavourable conditions. The data were statistically tested and it was found that a high degree of correlations existed between pH and oxygen consumption (r = 0.97). The analysis of covariance showed a significant relationship between oxygen consumption and pH (P < 0.05). Meanwhile, minimal variation in oxygen consumption was recorded between 30 and 40oC, with a
gradual decrease in dissolved oxygen concentration up to 12 hours of experimental time. At 50oC,
almost all dissolved oxygen was consumed by the larvae. The rate of oxygen consumption between
30 and 40oC was low during the first 9 hours of the experiment but it was significantly increased at later hours. A sudden increase in the oxygen consumption was recorded at 50oC, suggesting that it
might be the most unfavourable temperature condition. Meanwhile, a significant relationship was
observed between temperature and oxygen consumption (P < 0.05; r = 0.98).
Horseshoe crabs are said to be declining worldwide. However, there is still no published report on the status of horseshoe crabs in Malaysia. Thus, we report here eight informative microsatellite markers that were developed using the 5'-anchored ISSR-PCR enrichment procedure to diagnose the population genetic structure of the mangrove horseshoe crab, Carcinoscorpius rotundicauda from Peninsular Malaysia. This set of markers was tested on 127 samples and showed polymorphism in this species. Hence they should be useful in future essential population genetic studies of these living fossils in the Southeast Asian region.
<b>Background and Objective:</b> Horseshoe crabs are widely used in both traditional and modern pharmaceutical applications. Most of the previous studies on horseshoe crabs focused on their blood which contains hemolymph and amoebocyte lysate. This study aimed to determine the potential antibacterial and antifouling properties of different extracts from the carapace and the book gills of <i>Carcinoscorpius rotundicauda</i>. <b>Materials and Methods:</b> The crude extracts were subjected to the bioactivity tests using the disc-diffusion and the inhibition of biofilm-formation measurement assays, for both the antibacterial and antifouling activities respectively. <b>Results:</b> The results obtained indicated that the carapace extracts had stronger antibacterial and antifouling effects compared to the book gills extracts. Extracts obtained from the male displayed more activity compared to the extracts from the female with a few exceptions. Methanol and acetone carapace crude extracts showed the best overall performance. A sterol compound was isolated from the carapace acetone extracts of the male of <i>C. rotundicauda</i>. However, the compound did not display strong activity compared to the crude extract. The compound might be contributing to the observed activity with other components through a synergistic effect. <b>Conclusion:</b> The presence of antibacterial and antifouling activities in the carapace and book gills extracts could be added to the complexity of the defence mechanisms of horseshoe crabs. The results of this study, therefore, may contribute to the knowledge of the defence mechanisms of <i>C. rotundicauda</i>. Further research is needed to determine the bioactivities of other parts of the animal and to explore their potential applications.
The data available in this repository were gathered from Balok, the only most productive spawning site for horseshoe crabs Tachypleus gigas and Carcinoscorpius rotundicauda in East Coast of Peninsular Malaysia. The mangrove horseshoe crab, C. rotundicauda population and spawning data are available in the first table. The horseshoe crabs were retrieved from Balok River using 11.43 cm mesh size gill nets installed at the river mouth, the confluence and last meander. The arthropods were inspected for damage, abnormality and growth before their release into Balok River, particularly at the site of capture. Sediment samples were retrieved at their spawning grounds to ascertain sediment composition and size classifications which were also processed using Logarithmic Method of Moments. Water parameters like temperature, pH and salinity were also investigated during year 2016. All these information are compiled into the second table and arranged according to the period of data availability. The horseshoe crab catch data of years 2012, 2013, 2014, 2015 and 2016 were made available by artisanal fisher and compiled in the third and fourth table for inter-species comparison.
Carcinoscorpius rotundicauda and Tachypleus gigas may co-exist and share common spawning grounds elsewhere but at Balok (East Coast of Peninsular Malaysia), C. rotundicauda is an understudied species. Neglected as research candidate because of inaccessible spawning grounds, smaller size and less commercial value than T. gigas and also, difficulty to attain from the wild has made C. rotundicauda population status remaining unidentified at Balok. This standpoint drove the present attempt because anthropic activities like structure placement and mining are point-source for runoffs that load sediments into Balok River. While erosion-accretion events have altered Balok River width, the shore sediments in Balok Beach were transitioned between medium-fine and fine sand between years 2012 and 2016. Eventually by year 2016, the C. rotundicauda were depositing 5117 eggs in 91 nests from 200 to 1000 m range along this corridor facing South China Sea. From this yield, C. rotundicauda released 2880 eggs in 56 nests during the Southwest monsoon, 1254 eggs in 19 nests during the Northeast monsoon and 983 eggs in 16 nests during the Inter-monsoon seasons. Though female C. rotundicauda opted to lay their eggs in shallow burrows at lower shorelines, the absence of erosion and substantial silt and clay (>20%) deposition facilitates C. rotundicauda embryogenesis with brief periods of temperature and salinity shocks during day-time falling tides. This encourages C. rotundicauda to emerge with increasing abundance and carry out bi-monthly spawning at Balok Beach. In short, shore restoration initiatives like systematic boat docking, proper disposal of nets and waste and, periodic fish-catching operations were effectively led by the Balok fisher citizen scientist. This successful community joint-cooperation proves that citizen-led caretaking of degraded beaches offers marine life protection and are practical for coastal area management especially at areas where other oviparous animals such as turtles and crocodiles are harboured.
Perivitelline fluid, extracted from the fertilized eggs of horseshoe crabs, has been reported to play a
vital role in supporting embryogenesis as well as cell proliferation. The present study aims to evaluate the effect
of PVF on the expression of COL1A1 in human dental pulp stem cells (DPSCs). The cells were grouped into two;
untreated (control) and treated with a single dose of PVF (0.019 mg/ml). Gene expression was quantified for
COL1A1 on day 1, 3 and 7 using reverse transcriptase PCR. The expression of COL1A1 on day 3 of treated
group with PVF was the highest though there was a decline of COL1A1 expression on day 7. Mann Whitney test
was utilized to determine the significance of COL1A1 expression between treated and untreated groups.
Significant difference in the expression of COL1A1 was observed between the treated and untreated groups on
day 3 though there was no significance in the expression on day 7. The present study indicates that PVF may
have the potential to increase cell proliferation in human DPSCs.