Analysis of porcine DNA in several food products

Species identification in food products has grown interest in recent years since these foodstuffs are liable targets for fraudulent labelling. Consumers of Halal products typically buying those with a Halal logo stamped on the packaging without doubt for its authenticity. DNA-based methods on PCR amplification were used to identify the presence of porcine DNA in the processed food samples. Genomic DNA of porcine was extracted using DNA-Sorb-C and subjected to PCR amplification targeting the mitochondrial 12S rDNA gene and porcine leptin gene (LEP). Out of 66 samples, 37 samples showed amplified product of 387 bp with mitochondrial 12S rDNA gene and 59 samples showed amplified products of 152 bp with LEP. The verification for Halal authentication of food products is significant for economical, religious and public health concerns

Acute toxicity of malathion, dichlorvos and temephos in climbing perch (Anabas testudineus)

Malathion, dichlorvos and temephos are used globally to control a wide range of invertebrate pests especially in Malaysia. These pesticides usually enter aquatic environment by direct application or through overspray, runoff and watersheds. However, applications of pesticides may cause adverse impacts to many non-target organisms such as fish. The objective for this study was to determine the 96 hours lethal concentration (LC50) of each pesticide in climbing perch, Anabas testudineus. A total of 130 A. testudineus was subjected to 13 aquariums. Fish were exposed to different concentrations of each pesticide for 96 hours. Fish were observed daily and dead fish were removed immediately. The 96h LC50 value for malathion, dichlorvos and temephos was determined as 0.25 mg/L, 2.35 mg/L and 25.0 mg/L respectively. The results obtained were based on the probit analysis method as described by Finney 1952. From the values obtained, malathion, dichlorvos and temephos can be classified as highly toxic pesticides since it can kill 50 percent of the population even in lower concentration. Thus, the information in this study can be used as a guide to help environmental management to assure the effective use of these pesticides and to prevent indiscriminate use of pesticides

Effect of different thawing temperature and time on the viability of cryopreserved bovine spermatozoa / Farida Zuraina Hj Mohd Yusof and Syarifah Faezah Syed Mohamad

Cryopreservation is a technique in order to preserve cell for long time storage. It is widely used in agricultural field to store male gametes in the use of liquid nitrogen. In this study, three adult bulls breed Piedmontase were used. Semen collected from these bulls was then grouped into two; treatment group was subjected to magnetic activated cell sorting system (MACS) and being cryopreserved, whilst the control group was directly cryopreserved without MACS. Sperm were incubated in Annexin V before placed in the column attached to the magnetic stand. Semen analysis was done by using the Computer Assisted Sperm Analysis software (CASA). Parameters included are sperm velocity (VAP, VCL, VSL; um/s) and sperm progression (WOB, LIN, STR; %). For sperm viability it was determined by eosin-nigrosin staining technique. Effects of different thawing temperature and time were analysed by using iQ™SYBR® Green assay for qPCR with GAPDH as reference gene. Cryocapacitation like damage was then identified by the chlorotetracycline/Hoechst staining assay under blue-violet illumination of fluorescent microscopy (excitation: 400-440nm, emission: 470nm). All statistical data were analysed using SPSS version 14; p-value was set at 0.05 as significant level. Findings revealed that thawing at 37°C for 13 seconds for MACS processed sample was superior compared to other thawing procedures. At 37°C, it produced significant data of VCL (xm/s, expression of HSP70 gene and CTC/Hoechst assay with p<0.05. As a conclusion, usage of MACS preceding cryopreservation can enhance cell survival by thawing at 37°C for 30 seconds

Analysing population structure of Elaeis Oleifera germplasm using model-based approach programme STRUCTURE / Wan Nurhayati Wan Hanafi …[et al.]

Elaeis oleifera serves as a source of genetic foundation in oil palm improvement programme, as it possess several interesting agronomic traits such as slow growth, higher oil unsaturation and disease resistance. Malaysian Palm Oil Board (MPOB) has developed a collection of simple sequence repeats (SSRs) from Elaeis oleifera genome (E. oleifera-gSSRs). A total of 21 polymoprhic SSR markers were evaluated in the attempt to assess the population structure of E. oleifera populations. The appropriate common ancestry (K) value was determined to be seven from the likelihood scores. The profile from STRUCTURE analysis indicates considerable sharing of genetic components among E. oleifera population with an exception for Population 01 from Columbia and Population 02 from Costa Rica. The present study provides information on population structure of MPOB E. oleifera collection via model-based method for germplasm conservation and utilisation in breeding programmes

Isolation and characterization of polyhydroxyalkanoate (PHA)-producing, cellulolytic bacteria from municipal wastewater

Polyhydroxyalkanoates (PHA) are linear bio-esters produce in nature by certain microorganisms particularly when subjected to limited nutrients sources such as nitrogen while carbon sources are at abundance. PHA has attracted tremendous amount of attention from scientists globally due to their biodegradable property as environment-friendly alternative to petrochemical based plastics. However, the biggest drawback in using PHA is their high production cost. The aims of this study are to assess and characterise PHA-producing bacteria from municipal wastewater and compare their efficiencies in PHA production. A total of 34 out of 282 bacterial isolates from wastewater showed the ability to produce PHA using Sudan Black B staining. These isolates were then subjected to cellulase activity screen using carboxymethyl cellulose (CMC) agar. Two out of the 34 isolates were showed promising cellulose degrading capability and the cellulolytic activities were studied at 24, 48 and 72 hours of incubation respectively. The PHA production efficiencies of both isolates UiTM-E1 and UiTM-E2 were then compared using mineral salt media (MSM) supplemented with 1% and 2% glucose at 24, 48 and 72 hours of incubations respectively. Isolate UiTM-E2 showed the highest PHA production at 36.93% of its cell-dry weight (CDW) at 48 hours incubation

Bioactivity of clitoria ternatea crude extracts against pathogenic bacteria

Clitoria ternatea, sometimes referred to as the Asian pigeon wings blue pea, the butterfly pea, or the Darwin pea, is a Fabaceae plant species that has been shown to possess antibacterial effects against several pathogenic microbes. Hence, the present study has been carried out to access the antibacterial activity of C. ternatea flower extracted with water and methanol against pathogenic bacteria. The well and disk diffusion assays were performed to determine the antibacterial activity of C. ternatea flower extracts. The efficacy of the extracts was then evaluated via broth microdilution assay to obtain MIC and MBC values and the growth reduction assay. Meanwhile, the DPPH scavenging test was used to assess the antioxidant activity of the crude extracts. The results of the well and disc diffusion assays showed that Gram-positive bacteria were more sensitive to both extracts compared to Gramnegative bacteria. Meanwhile, the methanolic extract showed higher antibacterial activity on both Gram-positive and Gram-negative bacteria compared to the aqueous extract. The results of the MIC and MBC tests showed that the methanolic extract was bactericidal to both Gram-positive and Gram-negative bacteria. The aqueous extract, however, demonstrated bacteriostatic activity against Gram-negative bacteria and bactericidal activity solely against Gram-positive bacteria. After a 24-h exposure period, a growth reduction assay showed that the methanolic extract could suppress both Gram-positive and Gram-negative bacteria by up to 99%. Meanwhile, the aqueous extract showed an inhibition percentage value ranging from 75% to 96% after an incubation period. The aqueous extract had the lowest antioxidant activity, with an EC50 value of 87.78 µg/mL, whereas the methanolic extract had a fair amount of antioxidant activity when compared to the control (quercetin), according to the DPPH scavenging assay. The present study suggests that C. ternatea extracts as a potential antibacterial agent against pathogenic bacteria with significant antioxidant activity and this activity may be due to the presence of anthocyanin and its derivatives

Aqueous Extract of Clitoria ternatea Attenuates the Growth of Streptococcus mutans

In the human oral cavity, Streptococcus mutans is often observed and is a major contributor to tooth decay. Increased S. mutans levels may be linked to progressively more severe forms of periodontal disease because root exposure in people with periodontitis increases caries rates. Hence, a new potential antibacterial compound needs to be searched to combat this pathogenic bacterium. The butterfly pea, or Clitoria ternatea is an ornamental plant that has been reported to exhibit antibacterial properties against several bacteria. Thus, the goal of this investigation was to determine how well C. ternatea aqueous (CTA) extract inhibited S. mutans. The disk diffusion assay was performed to access the antibacterial properties of the CTA extract. The efficiency of the extract against the test bacterium was then determined through MIC/MBC determinations and a time-kill study. Meanwhile, the toxicity of the extract was tested using a brine shrimp lethality assay (BSLA). The CTA extract demonstrated substantial antibacterial activity against the test bacterium at a concentration of 200 mg/ml, with a diameter of the inhibition zone of 13.4±0.4 mm, according to the disc diffusion assay. The aqueous extract’s MIC and MBC values were found to be 100 and 400 g/mL, respectively. Time-kill analysis revealed the CTA extract exerted a strong bactericidal effect on S. mutans and this activity was dose-dependent. A scanning electron microscope (SEM) exhibited the bacterial cells experienced severe damage after being exposed to CTA extract including formation cavities, irregular shape, and crumpled cells. Thus, the present study suggested the potential of CTA extract as an antibacterial agent against oral cavity bacteria and can be used in the formulation of natural mouthwash due to no toxicity effect

Quantification of HSP70 gene expression and determination of capacitation status of magnetically separated cryopreserved bovine spermatozoa at different thawing temperature and time

The role of heat shock protein in reproduction is widely known as a molecular chaperone in aiding and repairing protein formation when stress occurred. The present objectives were to evaluate the effect of different thawing temperature and time on the expression of HSP70 gene expression and the capacitation status in cryopreserved bovine spermatozoa. Briefly, fresh ejaculates were obtained from three different adult bulls. The semen then underwent a sperm washing technique known as Magnetic Activated Cell Sorting System (MACS) and later on, cryopreserved. The sperm- containing straws were then thawed at five different thawing temperatures and time post-cryostorage; 20°C for 13 s, 37°C for 30 s, 40°C for 7 s, 60°C for 6 s and 80°C for 5 s. The RNA was extracted from each of the sperm’s pellets and converted to cDNA prior to the qPCR process. Capacitation status was then determined by means of CTC assay. The results showed that after the process of amplification, there is a significant different of HSP70 gene expression in MACS process samples when the thawing process was performed at 37°C for 30 s, with p<0.05. Furthermore, the CTC assay also showed that thawing at the same temperature gave less capacitated spermatozoa with p<0.05. As a conclusion, MACS yield spermatozoa with a better expression of HSP70 gene and less capacitated spermatozoa when thawing was done at 37°C for 30 s

Silk and silk-like fibres: halal attributes from a scientific viewpoint

The prophet Muhammad S.A.W. has made silk unlawful for the Muslim males. The development of silk industry around the globe had made abundant the availability of textile merchandise of silk origins. Advancement of technology created textile materials of surface properties, texture and ‘feel’ which mimicked pure silks. As such, consumers and contemporary Muslims could no longer differentiate between pure silk and synthetic silk-like fabrics and are forced to face the issues of halal textiles/fabrics for wear and clothing. This paper gives an insight and review of the characteristics and properties of silk and silk-based textiles in the scientist’s viewpoint. Ways and techniques to produce synthetic and composite fabrics which resembled pure silk as reported in the literatures are revealed and discussed to identify possibilities of segregating between pure and synthetic silk-like fibres. In conclusion, it is the purpose of this paper to expose scientific background and knowledge on silk and silk-like textiles to the community so as to gain a better understanding and perspective on the issues of halal textiles

Polymorphism of 11 Y Chromosome Short Tandem Repeat Markers among Malaysian Aborigines

The conventional technique such as patrilocality suggests some substantial effects on population diversity. With that, this particular study investigated the paternal line, specifically Scientific Working Group on DNA Analysis Methods (SWGDAM)-recommended Y-STR markers, namely, DYS19, DYS385, DYS389I/II, DYS390, DYS391, DYS392, DYS393, DYS438, and DYS439. These markers were tested to compare 184 Orang Asli individuals from 3 tribes found in Peninsular Malaysia. As a result, the haplotype diversity and the discrimination capacity obtained were 0.9987 and 0.9076, respectively. Besides, the most diverse marker was DYS385b, whereas the least was DYS391. Furthermore, the Senoi and Proto-Malay tribes were found to be the most distant, whereas the Senoi and Negrito clans were almost similar to each other. In addition, the analysis of molecular variance analysis revealed 82% of variance within the population, but only 18% of difference between the tribes. Finally, the phylogenetic trees constructed using Neighbour Joining and UPGMA (Unweighted Pair Group Method with Arithmetic Mean) displayed several clusters that were tribe specific. With that, future studies are projected to analyse individuals based on more specific sub-tribes