Studies on the storage stability of fermented red dragon fruit (Hylocereus polyrhizus) drink

The objective of this work was to study the effect of storage temperatures and duration on the stability of fermented red dragon fruit drink (FRDFD) on its betacyanins content, physicochemical and microbiological qualities (BPM) and determining sensory acceptability. Results showed that both storage temperatures and duration have a significant effect on betacyanins content and physicochemical properties of FRDFD. Aerobic mesophilic and yeast and mold counts were lower than 1 × 103 CFU/mL for FRDFD stored at both temperatures. The loss of betanin (16.53–13.93 g/L) at 4 °C was 15.73% with no significant changes in physicochemical properties from week two onwards compared to 56.32% (16.53–7.22 g/L) of betanin loss at 25 °C. At week eight, FRDFD stored at 4 °C still contained 13.93 g/L betanin with a pH value of 3.46, suggested its potential as a functional drink which is sensory acceptable (mean score > 80% using hedonic test) among consumers

Study on bioaccessibility of betacyanins from red dragon fruit (Hylocereus polyrhizus)

Betacyanins are bioactive dietary phytochemicals which can be found in red dragon fruit (RDF). Therefore, the bioaccessibility of betacyanins that present in fermented red dragon fruit drink (RDFD) and pressed red dragon fruit juice (RDFJ) was accessed in simulated gastric and intestinal digestion. Results disclosed that betacyanins from RDFD and RDFJ suffered minor loss (< 25%) at gastric-like environment but greater loss was observed during the intestinal phase digestion. After subjected to intestinal digestion, RDFD retained 46.42% of betanin while RDFJ retained 43.76%, with betanin concentration of 17.12 mM and 12.37 mM, respectively. Findings also revealed that RDFD exhibited higher antioxidant capacity compared to RDFJ after subjected to intestinal digestion, with values of 0.88 mM Trolox equivalent antioxidant capacity (TEAC) and 0.85 mM TEAC, respectively. The data suggests that betacyanins that present in RDF are bioaccessible while fermentation able to enhance the bioavailability with more betacyanins retained after intestinal digestion

Fermentation of red dragon fruit (Hylocereus polyrhizus) for betalains concentration

Red dragon fruits (RDF) contain high levels of health-promoting betalains but its bioavailability in plasma is low (< 1.0%). Therefore, fermentation technique was adopted to improve the yield of betalains presenting in fermented red dragon fruit drink (FRDFD) via concentration. Fermentation of RDF was carried out with autochthonous strains. The objectives of present study were to evaluate 1) the effects of fermentation duration 2) white refined cane sugar to flesh ratio and 3) types of sugar on betalainic (betanin, isobetanin) and non-betalainic phenolic compounds in FRDFD using HPLC-DAD, total phenolic content (TPC) and total flavonoid content (TFC) assay. Results indicated that all fermentation parameters showed a significant effect (p < 0.05) on the yields of betalainic (betanin, isobetanin) and non-betalainic phenolic compounds in FRDFD. The best fermentation parameters were 7 days fermentation at 10% white refined cane sugar to flesh ratio. The highest concentration of betanin, TPC and TFC achieved were 131.68 g/L, 1136.85 mg GAE/100mL and 10.39 mg CE/100mL respectively. The concentration of betanin obtained through best fermentation parameters (131.68 g/L) in present study has increased nine-fold compared to non-optimized fermentation (14.23 g/L). This indicated that fermentation is a potential economic processing technique to concentrate bioactive compounds present in functional drinks

Development of species-specific SCAR markers for identification and authentication of three rare Peninsular Malaysian endemic Coelogyne (Orchidaceae) orchids

Background: Coelogyne kaliana, Coelogyne stenochila and Coelogyne tiomanensis are three valuable rare orchid species endemic to Peninsular Malaysia, currently rampantly traded illegally via the internet and through local nurseries, which label them as hybrids to avoid enforcement detection. Drastic measures to ensure the continued existence of their populations in the wild should be introduced as they are rapidly diminishing into extinction, including the development of rapid and accurate species-specific identification tools. These three orchid species are highly similar morphologically and currently it is impossible to distinguish among them without their reproductive structures. Methods: RAPD-based species-specific SCAR markers were developed to distinguish and authenticate the identity of these three endemic Peninsular Malaysian Coelogyne species. Results: Three SCAR markers were successfully developed in this study. SCAR marker primer pair , CKL_f / CKL_r was specific to C. kaliana as it produced a unique single band of 271 bp but not in C. stenochila and C. tiomanensis. SCAR marker primer pair CST_f / CST_r amplified a single band of 854 bp in C. stenochila and two bands of different sizes (372 bp and 858 bp) in C. tiomanensis, but no amplification in C. kaliana. The third SCAR marker primer pair, CTI_f / CTI_r produced a single band (about 500 bp) for both C. stenochila and C. tiomanensis, but showed no amplification in C. kaliana. Conclusions: Although not all these SCAR markers were species amplification specific, they could be used to discriminate among the three Coelogyne species effectively. Accurate species identification is one of the most important steps to allow a proper management plan to be established in the effort to conserve these three endangered orchid species of Peninsular Malaysia. Besides, it could effectively put a stop to the illegal trading of these rare endangered orchid species worldwide

Phylogenetic relationships of the orchid genus Coelogyne in Peninsular Malaysia inferred from morphological characteristics and Internal Transcribed Spacer (ITS) sequence data

The phylogenetic relationships among the Peninsular Malaysian orchid genus Coelogyne were studied by morphological characteristics and sequence data of the internal transcribed region (ITS) from the nuclear ribosomal DNA (nrDNA). Coelogyne is a large genus of about 200 species distributed in pantropical areas from the Himalayas, Sri Lanka, India, Southern China and throughout South East Asia to Papua New Guinea. The widely accepted previous classification system was exclusively based on floral morphology. There were very few molecular systematic studies of Coelogyne done in Peninsular Malaysia thus far. In this study, 59 Coelogyne samples were collected throughout Peninsular Malaysia and 57 of them were identified to the species level. To study the phylogeny of this genus, morphological characters were utilized together with molecular evidences to generate the systematic hypotheses. Cluster analysis was performed using both the vegetative and floral characters. The results showed that three sections of Peninsular Malaysian Coelogyne, namely Longifoliae, Speciosae, and Fuliginosae were sister groups which were more closely related by forming one clade by itself. Another clade consisted of four other sections, namely Flaccidae, Coelogynae, Tomentosae, and Verrucosae. Molecular phylogenies obtained by using the Neighbour-Joining method showed the close relationship between the sections Tomentosae and Verrucosae, whereas usage of the Maximum Likelihood method demonstrated that three sections, namely Longifoliae, Speciosae, and Fuliginosae, were sister groups since they formed a single clade

Efficient and high-quality RNA isolation from metabolite-rich tissues of Stevia rebaudiana, an important commercial crop

Stevia rebaudiana, a perennial herb native to northeastern Paraguay, has gained immense attention globally over the recent decades due to the natural sweetness of its leaves. Like in most plants, this particular species contains high amount of secondary metabolites, thus rendering the isolation of high quality and quantity RNA extract for molecular applications rather challenging. An effective, high-yield and high-quality RNA isolation protocol for this economically important plant species was devised here based on the cetyltrimethylammonium bromide (CTAB) extraction method, with an additional genomic DNA (gDNA) removal step. DNA and other contaminants that may affect downstream applications were effectively removed. Our results exhibited that RNA samples isolated from the leaves and stems of Stevia rebaudiana using this improvised method are high in integrity and quality with RNA integrity number (RIN) of more than 8 and low in contaminants

Identification of simple sequence repeats (SSR) loci in Vanda Mimi Palmer expressed sequence tag database as potential marker(s) suitable for screening fragrance-related vandaceous and non-vandaceous orchids

Vanda Mimi Palmer (VMP) was largely cultivated for its intoxicating fragrance, huge commercial value and potential. It is a hybrid produced by crossing Vanda Tan Chay Yan and Vanda tessellate. Since fragrance characteristics of vandaceous orchids could not be determined based on the vegetative parts when the flowers are absent, our study is targeting on producing markers to detect their fragrance characteristics without the inflorescence. In our previous study, 2132 ESTs were generated from the VMP floral cDNA library. A total of 2,195 primer pairs were designed based on the mined EST-SSRs and primer pairs with amplicon size ranging from 100bp to 400bp and SSRs with three to five repeats were selected. A total of hundred and fifty primer pairs were chosen for PCR optimization performed using genomic DNA of VMP. Only 39 primer pairs were successfully optimised showing single banding pattern on 7% polyacrylamide gel (PAGE) and sent for sequencing. Three amplicons (obtained using the three primer pairs designated as P106, P117 and P140) showed the presence of the expected SSR. The primers were tested on genomic DNA extracted from other fragrant vandaceous and non-vandaceous orchids, and successfully amplified identical target region as in VMP. The presence of highly conserved and similar EST-SSRs sequences might serve as evidence of structural conservation and relationships across orchid species. Hence, we suggest these three primer pairs can be utilized to detect other vandaceous and non-vandaceous orchids

First molecular detection of porcine bocavirus in Malaysia

Several strains of porcine bocaviruses have been reported worldwide since their first detection in Sweden in 2009. Subsequently, the virus has been reported to be associated with gastrointestinal and respiratory signs in weaner and grower pigs. Although Malaysia is host to a self-sufficient swine livestock industry, there is no study that describes porcine bocavirus in the country. This report is the first to describe porcine bocavirus (PBoV) in Malaysian swine herds. PBoV was identified in various tissues from sick and runt pigs using the conventional PCR method with primers targeting conserved regions encoding for the nonstructural protein (NS1) gene. Out of 103 samples tested from 17 pigs, 32 samples from 15 pigs were positive for porcine bocavirus. In addition, a higher detection rate was identified from mesenteric lymph nodes (52.9%), followed by tonsil (37.0%), and lungs (33.3%). Pairwise comparison and phylogenetic analyses based on a 658-bp fragment of NS1 gene revealed that the Malaysian PBoV strains are highly similar to PBoV3 isolated in Minnesota, USA. The presence of porcine bocavirus in Malaysia and their phylogenetic bond was marked for the first time by this study. Further studies will establish the molecular epidemiology of PBoV in Malaysia and clarify pathogenicity of the local isolates

Annotated checklist of orchids found in Merapoh Trail (Gunung Tahan, Malaysia)

Gunung Tahan is the highest mountain peak in Peninsular Malaysia (2187 m a.s.l.) and is a part of Taman Negara Pahang that houses many rare and endemic species of flora and fauna. This account is conducted to evaluate, compile, and update the orchids collections from Gunung Tahan after almost 126 years, since the first botanical exploration was conducted by H.N. Ridley and other foreign botanist. The compilation presented in this study resulted in 294 species obtained from the selected herbarium, published record, and current expedition. A total of 57 species were recorded for recent work, out of which 18 species were new discoveries for Merapoh Trail and thus reported as new records for Gunung Tahan. IUCN Red List of Threatened Species version 2017.3 listed two species; Claderia viridiflora Hook.f. and Spiranthes sinensis (Pers.) Ames as least concern species and Paphiopedilum bullenianum (Rchb.f.) Pfitzer as endangered species. The list of the compiled species from herbarium and published records for Gunung Tahan is also included

First molecular detection of porcine bocavirus in Malaysia

Several strains of porcine bocaviruses have been reported worldwide since their first detection in Sweden in 2009. Subsequently, the virus has been reported to be associated with gastrointestinal and respiratory signs in weaner and grower pigs. Although Malaysia is host to a self-sufficient swine livestock industry, there is no study that describes porcine bocavirus in the country. This report is the first to describe porcine bocavirus (PBoV) in Malaysian swine herds. PBoV was identified in various tissues from sick and runt pigs using the conventional PCR method with primers targeting conserved regions encoding for the nonstructural protein (NS1) gene. Out of 103 samples tested from 17 pigs, 32 samples from 15 pigs were positive for porcine bocavirus. In addition, a higher detection rate was identified from mesenteric lymph nodes (52.9%), followed by tonsil (37.0%), and lungs (33.3%). Pairwise comparison and phylogenetic analyses based on a 658-bp fragment of NS1 gene revealed that the Malaysian PBoV strains are highly similar to PBoV3 isolated in Minnesota, USA. The presence of porcine bocavirus in Malaysia and their phylogenetic bond was marked for the first time by this study. Further studies will establish the molecular epidemiology of PBoV in Malaysia and clarify pathogenicity of the local isolates