First Report on Wild Ginger (Family: Zingiberaceae) Species Composition with New Records in Limestone Forests of Kelantan, Peninsular Malaysia.

The wild gingers in the family Zingiberaceae have a wide range of habitat distribution. The species growing in Malaysian forests are the most studied. Nevertheless, the aromatic perennial herb family found in limestone forests is the least studied. The present study identified the ginger species compositions, determined the conservation status of the identified ginger species, and compared the distribution of the ginger species in selected limestone forests of Kelantan due to the lack of intensive study focusing on wild gingers in Malaysian limestone forests, especially in the state of Kelantan, to date. In various months, wild ginger species observation was conducted at four limestone forests in Kelantan. From the survey performed during the present study, Gua Setir and Gua Ikan recorded 16 species with 12.5% overlapping species. Gua Setir comprised 61.5% more ginger species than Gua Ikan. In total, 13 species (81.25%) were evaluated based on the Red List of Threatened Species by the International Union for Conservation of Nature (IUCN). Three wild ginger species listed as high conservation value (HCV), Zingiber aurantiacum, Zingiber petiolatum and Zingiber wrayi, were identified at the limestone karst valley of Gua Setir. The current study presented updated and new records of the limestone wild ginger flora in Kelantan. The research also demonstrated that each limestone forest consisted of different combinations of ginger species. Consequently, conservation efforts and sustainable management currently enforced in the limestone forests would lead to long-term protection of the plants. Furthermore, the wild gingers could become a tourist attraction for limestone forests located in recreational areas.

Halia hutan (Famili: Zingiberaceae) mempunyai taburan habitat yang luas. Kajian mengenai tumbuhan herba yang berbau harum ini kurang dilakukan di kawasan hutan batu kapur Malaysia. Kajian ini telah mengenal pasti komposisi spesies halia, menentukan status pemuliharaan spesies halia yang telah dikenal pasti, serta membandingkan taburan spesies halia di beberapa hutan batu kapur di Kelantan. Kajian ini juga dilakukan kerana kekurangan kajian intensif yang memfokuskan halia hutan di hutan batu kapur Malaysia, terutamanya di negeri Kelantan, sehingga kini. Pemerhatian terhadap spesies halia hutan telah dilakukan di empat hutan batu kapur di Kelantan. Gua Ikan dan Gua Setir telah merekodkan 16 spesies halia semasa pemerhatian tersebut dengan 12.5% spesies yang bertindih. Gua Setir merangkumi 61.5% spesies halia berbanding Gua Ikan. Secara keseluruhan, 13 spesies (81.25%) halia yang dikenal pasti telah dinilai berdasarkan Red List of Threatened Species oleh International Union for Conservation of Nature (IUCN). Sebanyak tiga spesies halia telah disenaraikan sebagai spesies high conservation value (HCV) iaitu Zingiber aurantiacum, Zingiber petiolatum dan Zingiber wrayi. Ketiga-tiga spesies ini telah dikenal pasti di lembah kars Gua Setir. Kajian ini telah memberikan rekod spesies yang telah dikemas kini serta rekod baru bagi spesies halia hutan di kawasan batu kapur di Kelantan. Kajian ini juga menunjukkan bahawa setiap hutan batu kapur terdiri daripada gabungan spesies halia yang berbeza. Oleh yang demikian, usaha pemuliharaan dan pengurusan mampan yang dilakukan pada masa kini di hutan batu kapur akan membawa kepada perlindungan jangka panjang untuk tumbuhan halia. Tambahan pula, tumbuhan halia hutan juga boleh dijadikan sebagai tarikan pelancong untuk hutan batu kapur di kawasan rekreasi.

Effect of Cryopreservation Method Supported with Biochemical Analyses in the Axillary Bud of Jewel Orchid, Ludisia discolor.

This study investigated conserving an endangered terrestrial jewel orchid Ludisia discolor, using in vitro grown axillary buds. Excised segments of axillary buds (4-5 mm in length) were precultured on a modified Murashige and Skoog (MS) medium supplemented with 0.2 M sucrose for 24 h and osmoprotected in a loading solution for 20 min. Then, axillary buds were dehydrated in plant vitrification solution 2 (PVS2) for 10 min at 0 °C and incubated in liquid nitrogen for 1 h. Subsequently, axillary buds were rewarmed rapidly by dilution solution and transferred to a growth recovery medium supplemented with 0.05 µM melatonin, which led to an improved survival chance (16.67%) for cryopreserved L. discolor. The osmotic stress and the overproduction of reactive oxygen species (ROS) during cryopreservation stages may result in cryoinjuries and poor survival as increased levels of proline (5.51 µmol/g), catalase (85.64 U/g), peroxidase (565.37 U/g), and ascorbate peroxidase activities (12.19 U/g) were detected after dehydration, preculture, rewarming, and loading stage, respectively. Results obtained from this study indicate that further experimental designs which apply different PVS and exogenous antioxidants are needed for improved survival and regrowth of L. discolor.

Proximate analysis and fatty acid of Corbicula fluminea (C. fluminea) tissue in Kelantan, Malaysia.

Corbicula fluminea (C. fluminea) is one of the mollusc species commonly eaten as a popular snack in Kelantan, Malaysia. This species contributes to the local economic activity. However, the handling process of C. fluminea at different processing stages (raw, smoke and selling stages) is believed to have affected the nutritional value in their soft tissue. Hence, this study aims to provide information about the chemical content (moisture, crude fat, ash, crude protein and carbohydrate) of C. fluminea and fatty acid of smoked C. fluminea at different processing stages. Samples were collected from Pasir Mas and Tumpat, Kelantan, Malaysia. The chemical content analysis was carried out based on the Association of Official Analytical Chemists (AOAC) standard procedures. The results have indicated that moisture (80.93 ± 0.37%) and crude fat (10.82 ± 0.21%) in C. fluminea were significantly higher (p < 0.05) at the raw stage. On the other hand, the ash (2.09 ± 0.03%) and crude protein (9.45 ± 0.42%) were significantly higher (p < 0.05) at smoked stage. Meanwhile, carbohydrate (3.39 ± 0.49%) was significantly higher (p < 0.05) at the selling stage. The fatty acid percentage of smoked C. fluminea indicates that the total saturated fatty acid (SFA) content was higher than the total of both monounsaturated fatty acids (MUFA) and polyunsaturated fatty acids (PUFA). The obtained data indicates that the composition percentage of omega-6 (n-6) in PUFA is significantly greater than omega-3 (n-3). The n-6/n-3 ratio of smoked C. fluminea is 2.95-3.11 and therefore can be considered as a healthy diet. The findings of this study can improve consumers' health through sufficient nutrient consumption and also provide economic benefits by increasing sellers' income in Kelantan.

Health Risk Assessment of Heavy Metals from Smoked Corbicula fluminea Collected on Roadside Vendors at Kelantan, Malaysia.

Corbicula fluminea serves as traditional food to the local people in Kelantan, Malaysia. Concerns regarding river contamination, smoking method, and associated adverse effects on public health had been increasing. Hence, this study aims to measure the level of heavy metals (Cd, Cu, Mn, Pb, and Zn) and assess human health risk in C. fluminea consumption at Kelantan. Heavy-metal analysis was done using flame atomic absorption spectrophotometry, while human health risk was assessed using provisional tolerable weekly intake (PTWI), target hazard quotient (THQ), and hazard index (HI). The estimated weekly intake (EWI) for all metals was found within PTWI, while THQ for Cd, Cu, Mn, Pb, and Zn was 0.12, 0.06, 0.04, 0.41, and 0.03, respectively. The HI was calculated at 0.61 which is less than 1, considered as the safe consumption level. Therefore, C. fluminea consumption in this study was found safe from the health risk of noncarcinogenic effect over a lifetime.

Antimicrobial activity of artemisinin and precursor derived from in vitro plantlets of Artemisia annua L.

Artemisia annua L., a medicinal herb, produces secondary metabolites with antimicrobial property. In Malaysia due to the tropical hot climate, A. annua could not be planted for production of artemisinin, the main bioactive compound. In this study, the leaves of three in vitro A. annua L. clones were, extracted and two bioactive compounds, artemisinin and a precursor, were isolated by thin layer chromatography. These compounds were found to be effective in inhibiting the growth of Gram-positive and Gram-negative bacteria but not Candida albicans. Their antimicrobial activity was similar to that of antibactericidal antibiotic streptomycin. They were found to inhibit the growth of the tested microbes at the minimum inhibition concentration of 0.09 mg/mL, and toxicity test using brine shrimp showed that even the low concentration of 0.09 mg/mL was very lethal towards the brine shrimps with 100% mortality rate. This study hence indicated that in vitro cultured plantlets of A. annua can be used as the alternative method for production of artemisinin and its precursor with antimicrobial activities.