Porcupine Bezoar Research Articles

The Trade of Porcupines in Malaysia With International Trade Links

Background and Research Aims Porcupines are frequently traded in Asia but evidence of this remains undocumented. This study was undertaken to address the dearth of information on porcupine trade dynamics using Malaysia as a case study. Methods To accomplish this, traditional Chinese medicine (TCM) outlets throughout the country were surveyed, the existence of porcupine farms were determined and porcupine seizures in Malaysia from 2011 – 2019 were analysed. Results The data revealed that porcupines are coveted predominantly for bezoars used in TCM and for breeding stock. Porcupine bezoars were found in 90% of TCM outlets surveyed though it is likely that the market consists of genuine and fake products. Porcupine farming appears to be gaining popularity and predominantly caters to a demand for breeding pairs. There were 47 seizures amounting to 110 porcupines of at least three species. Seizures included mostly live animals but also meat and quills. Conclusion The harvesting and consumption of porcupines is legally permitted in Malaysia. However, it is evident that illegal hunting of porcupines is also occurring. Captive breeding is unlikely to reduce poaching of porcupines considering the high value associated with wild-sourced bezoars. Local use is also driving international trade of bezoars particularly from Indonesia. Implications for Conservation Research on porcupine trade dynamics in Southeast Asia is limited. This is a conservation concern particularly as porcupines are valued as commercial commodities. As Asian porcupines face a multitude of synergistic threats ( i.e., habitat loss, human conflict and illegal hunting), understanding these cumulative impacts on wild populations is a conservation priority. More research on international trade dynamics is also warranted as it is currently occurring without any regulation or monitoring. Listing porcupines in the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES) could potentially mitigate this.

Valuable stones: The trade in porcupine bezoars

Old-world porcupines (Order: Rodentia Family: Hystricidae) face many threats, including an increasing demand for their different body parts such as meat, quills, hairs and bezoars. Bezoars are masses of undigested organic and inorganic material that are occasionally formed in an animal’s gastrointestinal tract. Bezoars of a variety of species, especially porcupines, have been used for medicinal purposes for centuries and have high commercial value. Demand for bezoars appears to have increased substantially in recent years.We monitored e-commerce websites in Singapore, Malaysia, and Indonesia to estimate the extent of porcupine bezoars for sale. We found 121 listings (excluding listings advertising supply ability instead of specific bezoars) advertising approximately 680–1332 bezoars and likely representing ten-fold as many porcupines within our three month study period. Most listings were from sellers located in Indonesia, followed by sellers from Malaysia, and Singapore. The mean adjusted price per gram was 151.8 USD, which is substantially lower than previously reported prices in the literature. Porcupines have an important ecological role and highly developed social structures. Current trade levels are likely unsustainable and we predict that porcupine species may become threatened in the future should current trade levels continue. We urge source and consumer countries to: (i) review the species’ conservation status in range countries; (ii) regulate domestic trade through legislative changes in countries where trade is most prominent; and (iii) ensure existing laws are enforced. Additionally, the Hystricidae may benefit from a CITES Appendix II listing to monitor and regulate the international trade.

Anticancer activity of grassy Hystrix brachyura bezoar and its mechanisms of action: An in vitro and in vivo based study

Porcupine bezoar (PB) is a calcified undigested material generally found in porcupine’s (Hystrix brachyura) gastrointestinal tract. The bezoar is traditionally used in South East Asia and Europe for the treatment of cancer, poisoning, dengue, typhoid, etc. However, limited scientific studies have been performed to verify its anticancer potential to substantiate its traditional claims in the treatment of cancers. Hence, this study was aimed at investigating the in vitro and in vivo anticancer properties of two grassy PB aqueous extract (PB-A and PB-B) using A375 cancer cell line and zebrafish model, respectively. This paper presents the first report on in vitro A375 cell viability assay, apoptosis assay, cell cycle arrest assay, migration assay, invasion assay, qPCR experimental assay and in vivo anti-angiogenesis assay using the grassy PBs. Experimental findings revealed IC50 value are 26.59 ± 1.37 μg/mL and 30.12 ± 3.25 μg/mL for PB-A and PB-B respectively. PBs showed anti-proliferative activity with no significant cytotoxic effect on normal human dermal fibroblast (NHDF). PBs were also found to induce apoptosis via intrinsic pathway and arrest cell cycle at G2/M phase. Additionally, the findings indicated its ability to debilitate migration and invasion of A375 cells. Further evaluation using embryo zebrafish model revealed LC50 = 450.0 ± 2.50 μg/mL and 58.7 ± 5.0 μg/mL for PB-A and PB-B which also exerted anti-angiogenesis effect in zebrafish. Moreover, stearic acid, ursodeoxycholic acid and pregnenolone were identified as possible metabolites that might contribute to the anticancer effect of the both PBs. Overall, this study demonstrated that PB-A and PB-B possess potential in vitro and in vivo anticancer effects which are elicited through selective cytotoxic effect, induction of apoptosis, inhibition of migration and invasion and anti-angiogenesis. This study provides scientific evidence that the porcupine bezoar do possess anti-cancer efficacy and further justifies its traditional utility. However, more experiments with higher vertebrae models are still warranted to validate its traditional claims as an anticancer agent.

A traditional folk medicine in Malaysia: porcupine bezoar

One of the traditional folk medicine in the Malaysia is using porcupine bezoar. It was reported as universal medicine in ancient time. This bezoar has been a myth to the locals. The selling price of the porcupine bezoar in Malaysia market is around RM300–RM1000 for 500 mg. The widespread use of the bezoar as an alternative medicine for treatment of deadly illnesses in Malaysia was passed down from generations of the multi-diverse ethnicity environment that existed here. Additionally, porcupine bezoar was also known as lifesaver and used by patients with terminal illness. Porcupine bezoar was found in the gastrointestinal tract of porcupine and made up of indigestible plant matters. Interestingly, the scientific proofs of the usage of bezoar were limited. The chemical profile and the medicinal properties of the bezoar is still unknown. The main focus of this review paper is on the porcupine bezoar which can be found in Malaysia.

Hystrix brachyura Bezoar Characterization, Antioxidant Activity Screening, and Anticancer Activity on Melanoma Cells (A375): A Preliminary Study.

Porcupine bezoars (PBs) are masses of undigested calcareous concretions formed within the gastrointestinal tract. There are undocumented claims that PBs have antioxidant activity and can treat cancers. However, limited scientific study has been carried out to verify these traditional claims. Hence, this study was conducted to characterize the chemical profile and validate the antioxidant and anticancer activity against melanoma cells (A375). PB extract was initially subjected to Fourier-transform infrared spectroscopy (FTIR), gas chromatography–mass spectrometry (GCMS), total phenolic content (TPC), and total flavonoid content (TFC) analyses. The bioautography of antioxidant assays, namely 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS), 2,2-diphenyl-1-picrylhydrazy (DPPH), and β-carotene was performed. An in vitro A375 cell viability assay, apoptosis assay, cell cycle arrest assay, and gene expression assay were carried out as well. The experimental finding revealed 5,10-diethoxy-2,3,7,8-tetrahydro-1H,6H-dipyrrolo[1,2-a:1′,2′-d]pyrazine, ursodeoxycholic acid, and cholest-5-en-3-ol (3 beta)-, carbonochloridate are major compounds detected in PB extract. PB extract has low phenolic content, viz. 698.7 ± 0.93 (µg GAE/5 mg dry weight). The bioautography antioxidant assays revealed a potent antioxidant effect (ABTS > DPPH > β-carotene), with free radical scavenging activity. Furthermore, PB extract exhibited dose- and time-dependent inhibition of cancer activity on A375 cells due to the exhibition of apoptosis via an intrinsic pathway.

Tannic acid-rich porcupine bezoars induce apoptosis and cell cycle arrest in human colon cancer cells

Background: Porcupine bezoar, a phytobezoar used as traditional medicine, was recently claimed to effectively treat cancer. However, there is a lack of scientific evidence to prove the claim. Objectives: This study aimed to scientifically examine porcupine bezoars as a potential anticancer agent and to investigate their principal bioactive constituents. Materials and Methods: The porcupine bezoars were extracted using methanol and further Sephadex LH-20 column chromatography was used to enrich the tannins content. The inhibitory effects of the crude extracts on a panel of cancer cell lines were first determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Then, the anticancer activities of the enriched fractions in selected cell lines were analyzed, while the chemical composition of the active fraction was identified using liquid chromatography––electrospray ionization-tandem mass spectrometry. Results: Crude extracts of black date and powdery date effectively inhibited colon cancer cell lines HT-29 and HT-116, but not the normal colon cells, and their tannin-enriched fractions demonstrated higher inhibitory effects when compared to the extracts. Further, the fractions arrested cell cycle at S phase and induced apoptosis in treated colon cancer cells with a similar effect to that of commercial tannic acid. Lipoxygenase activity which plays a role in tumorigenesis of colon cancer was also inhibited by these fractions. Chemical analysis found that both the enriched fractions and commercial tannic acid share similar chemical constituents, including gallic acid and its derivatives (polygalloyl glucose). Conclusion: Together, the results suggest that tannic acid in porcupine bezoars may inhibit colon cancer cells by interfering cell proliferation and triggering program cell death in the cells.

In Vitro Evaluation of Porcupine Bezoar Extracts as Anticancer Agent on A549 -A Preliminary Study

Porcupine bezoar (PB) was reported to possess medicinal properties in old medical manuscript. However, its potential as anticancer agent on human lung cancer cells (A549) is not yet studied. In present study, porcupine bezoar was tested to observe its ability in inhibiting cell growth of cancer cell (A549) and its cytotoxicity on Normal Human Gingival Fibroblast Cell (HGF-1). A549 cells morphology was observed after treated with bezoars for 72 hours. The ability of bezoars to induce DNA damage and apoptosis was analyzed by staining cells with Hoechst 33428(nucleus) and Rhodamine Phalloidin (f-actin). The A549 IC50 is 13.6±1.58μg/ml A549 growths was inhibited in dose-dependent pattern, but no inhibition found on normal HGF-1 cells. Treated A549 morphology shows sign of apoptosis such as DNA fragmentation, cytoplasm shrunk and vacuolation. The finding in this study suggests PB extracts able to inhibit cell growth, induce DNA damage and apoptosis, further analysis need to be done to verify the mechanism.

Antioxidant and Intracellular Reactive Oxygen Species/Reactive Nitrogen Species Scavenging Activities of Three Porcupine Bezoars from Hystrix brachyura.

Background:Porcupine dates are phytobezoar stones that are used in Traditional Chinese Medicine (TCM) treatments against cancer, postsurgical recovery, dengue fever, etc. The medicinal values have not been scientifically investigated due to the availability and high pricing of the dates.Objectives:This paper represents the first report on the phytochemical content, in vitro antioxidant and intracellular reactive oxygen species (ROS)/reactive nitrogen species (RNS) scavenging properties of the extracts of three porcupine dates: grassy date (GD), black date (BD), and powdery date (PD).Materials and Methods:Dried samples were extracted with methanol and lyophilized. Samples were screened for phytochemical constituents, in vitro antioxidant assays based on total phenolic content (TPC), free radical scavenging, and ferric reducing power (FRP) as well as intracellular ROS and RNS scavenging properties.Results:Phytochemical screening and total tannins assay revealed that tannins, cardiac glycosides, and terpenoids were found in all porcupine dates with tannins forming the major portion of the TPC. In comparison to GD, BD and PD were found to contain significantly high TPC, radical scavenging activity, and FRP. At 200 μg/ml, BD and PD remarkably scavenged 2, 2-azobis (2-amidinopropane) dihydrochloride-induced ROS in RAW264.7 cells and significantly reduced nitric oxide in lipopolysaccharide-stimulated cells.Conclusion:Overall, BD and PD exhibited promising in vitro antioxidant as well as intracellular ROS/RNS scavenging properties.SUMMARY Tannins, cardiac glycoside, and terpenoids were found in all three types of porcupine dates with tannins being the major compoundsAntioxidant contents and properties of three dates were in the order black date (BD) > powdery date (PD) > grassy dateBD and PD extracts showed significant intracellular reactive oxygen species and reactive nitrogen species scavenging properties. Abbreviations Used: TCM: Traditional Chinese Medicine, BD: Black date, GD: Grassy date, PD: Powdery date, TPC: Total phenolic content, FRS: Free radical scavenging, FRP: Ferric reducing power, NO: Nitric oxide, ROS: Reactive oxygen species, RNS: Reactive nitrogen species, GAE: Gallic acid equivalent, AAE: Ascorbic acid equivalent, PVPP: Polyvinylpolypyrrolidone, DCFH-DA: Dichloro-dihydro-fluorescein diacetate, AAPH: 2, 2-azobis (2-amidinopropane) dihydrochloride, LPS: Lipopolysaccharide