Extraction and Characterization of Litopenaeus vannamei’s Shell as Potential Sources of Chitosan Biopolymers

Chitin is the second most abundant polysaccharide,produced mainly as an industrial waste stream during crustacean processing.Chitin can be derived into chitosan through the deacetylation process.Conversion of shrimp waste into chitosan via the deacetylation process could be considered a practical approach for shell waste remediation.In this study,chitosan’s physicochemical characteristics extracted from two types of Pacific white leg shrimp,L.vannamei’s shell(i.e.,rough and smooth),were compared with commercial chitosan.The yield,moisture,ash,solubility,water and fat binding capacity were measured.The degree of deacetylation(DDA)was calculated using FTIR,and their chemical Structure was confirmed using XRD and SEM-EDS.Both extracted chitosan showed no significant difference in yield,moisture,ash,solubility and water binding capacity but showed a significant difference with commercial chitosan.Moreover,the fat binding capacity of commercial chitosan showed the lowest percentage(408.34±0.83%)as compared to extracted chitosan(smooth shell 549.59±12.48%;rough shell 500.55±12.10%).The DDA indicated that extracted chitosan from the smooth and rough shell was considered good chitosan as compared to commercial chitosan with 84.08±1.27%,80.78±0.79%and 74.99±1.48%,respectively.Additionally,the presence of hydroxyl and amino groups from FTIR and a good crystallinity index was recorded using XRD of extracted chitosan.Based on observed characteristics,shrimp shell waste from L.vannamei can achieve chitosan standard quality as a biopolymer and highly potential to be applied in various industrial applications.

Effects of Tinospora tuberculata leaf methanol extract on seedling growth of rice and associated weed species in hydroponic culture

The study was conducted to evaluate the responses of rice and rice weed seedlings(barnyardgrass and weedy rice) at the three-leaf stage to Tinospora tuberculata leaf methanol extract(3.12, 6.25 and 12.5 g L–1) under hydroponic culture. It shows that the leaf methanol extract had various degrees effects depending on target plant species and each tested index(biomass, root length, shoot length, transpiration volume, chlorophyll a, chlorophyll b and carotenoid contents). The effective concentration of the leaf extract capable of reducing 50% of rice growth was higher than those of target weed species. Moreover, the root length was more tolerant to leaf methanol extract in comparison to the other plant parameters measured. A greater reduction was observed in chlorophyll a content compared to chlorophyll b and carotenoid. The results revealed that the reduction of transpiration volume closely coincided with the magnitude of growth inhibition of tested plants. Ultra-fast liquid chromatography analysis revealed 11 of 32 peaks in chemical profile, including benzoic acid, caffeic acid, chlorogenic acid, isoorientin, isovitexin, orientin, p-anisic acid, syringic acid, trans-cinnamic acid, trans-ferulic acid, and vitexin have the same retention time with those peaks of the extract. The amount of compounds was present in the range of between 4 817 and 115.5 mg kg–1 dry weight(DW). The concentration-response bioassay of all 11 individual compounds and their equimolar mixture against the seeds of barnyardgrass revealed their contribution in the allelopahic activity of T. tuberculata leaf extract. The examined compounds and their combination exhibited various degrees of growth inhibitory effects on the early growth of barnyardgrass. Therefore, the specific number, concentration, combination and inhibitory activity of bioactive compounds leads to allelopathy activity of T. tuberculata leaves which could be employable directly as a natural herbicide and its growth inhibitor compounds can be used as a template for producing new herbicides.

Antibiotic resistance profile and RAPD analysis of Campylobacter jejuni isolated from vegetables farms and retail markets

Objective:To investigate antibiotic resistance profile and characterize Campylobacter jejuni(C.jejuni) isolates using random amplified polymorphic DNA(RAPD) analysis.Methods:Ninety eight C.jejuni isolates from farms and retail outlets were screened against 10 antibiotics commonly used clinically and agriculturally by using disk diffusion method.RAPD analysis was done to characterize 98 C.jejuni isolates.Results:Fifty-one percent of the isolates had multiple antibiotic resistance index 0.2 and below.This indicated that the isolates in the vegetables were not from the high risk environment or extensive farming practices.C.jejuni isolates found resistant towards penicillin G(93%),vancomycin(86%),ampicillin(35%),erythromycin(28%),gentamycin(4%),amikacin(3%),enrofloxacin(1%),norfloxacin(1%) and no resistance towards ciprofloxacin.RAPD clustering analysis showed that the contamination of C.jejuni in vegetables was likely due to cross contamination at retail markets.Conclusions:C.jejuni contamination in vegetables at retail markets was due to cross contamination.Current finding proved that C.jejuni in small scale vegetables production was less expose towards antibiotic abuse.

Phenolics, fatty acids composition and biological activities of various extracts and fractions of Malaysian Aaptos aaptos

Objective: To investigate phenolics, fatty acids composition and biological activities of various extracts and fractions of Malaysian Aaptos aaptos. Methods: Fatty acid methyl ester was analyzed by gas chromatography-flame ionization detector. Antioxidant activity was determined using 2,2-diphenyl-picrylhydrazyl radical scavenging assay and total phenolics content by Folin-Ciocalteu procedure. Vero cells viability was evaluated using methyl thiazole tetrazolium and the inactivation of herpes simplex virus type 1 by neutral red uptake assay. p-Hydroxybenzamide isolated by column chromatography was characterized by utilizing nuclear magnetic resonance spectroscopy and electron impact mass spectrometry. Results: The chloroform, ethyl acetate and methanol extracts of Aaptos aaptos produced higher portions of straight-chain saturated fatty acid, while hexane extract mainly consisted of unsaturated fatty acid. The five majors of fatty acid methyl ester were identified as behenic acid, cis-10-heptadecenoic acid and cis-10-pentadecenoic acids, palmitic acid and tricosanoic acid. In addition, among all organic extracts, chloroform extract inactivated herpes simplex virus type 1 while exhibited weak cytotoxic activity against normal Vero cells and also exhibited strong cytotoxic activity on HL-60, MCF-7, K562, CEM-SS and WEHI-3 B cells. A phenolic compound, p-hydroxybenzamide was also isolated from the sponge. Conclusions: Aaptos aaptos could be a source to derive the potential antiviral and anticancer agents. However, further studies are needed to determine the mechanism involved in the process.

The Effect of Age on Milk Yield and Milk Composition in Saanen Dairy Goats

The goals of this study were to determine the weekly milk production of Saanen goats at UniSZA Pasir Akar Farm,and to evaluate milk production and composition in three age groups.From January 2021 to May 2021,a study was conducted at the UniSZA Pasir Akar Farm in Besut,Terengganu.Ninety goat milk samples were collected and stored in sterile falcon tubes.Milk samples were stored in an insulated box at 5℃ before being transported to the laboratory and stored at-20℃ until further analysis.Milk samples were analyzed in four replicates for each sample group using the Milkotester.The one-way analysis of variance(ANOVA)method was used to analyze raw data among age groups,with p<0.05 indicating a significant difference.The highest milk yield was produced by four-year-old goats(943.9 g/d),followed by three-year-old goats(850.5 g/d)and two-year-old goats(571.1 g/d),respectively.Solid non-fat(SNF),protein,and lactose content showed the highest in percentage in the 3-year old group,at 7.80%,2.80%,and 4.27%,respectively.However,the fat content revealed an unusual pattern,with 2.87%,2.77%,and 3.33%representing 2,3 and 4 years old.This occurred due to other factors such as feed,breed,and age.In conclusion,this study found a significant difference in milk yield and composition across three age groups.However,this is only a preliminary result based on a small number of animals and a short study period.Future studies will perhaps,use larger sample sizes and parameters to validate the current result.

mRNA transcriptome profiling of human hepatocellular carcinoma cells HepG2 treated with Catharanthus roseus-silver nanoparticles

BACKGROUND The demand for the development of cancer nanomedicine has increased due to its great therapeutic value that can overcome the limitations of conventional cancer therapy.However,the presence of various bioactive compounds in crude plant extracts used for the synthesis of silver nanoparticles(AgNPs)makes its precise mechanisms of action unclear.AIM To assessed the mRNA transcriptome profiling of human HepG2 cells exposed to Catharanthus roseus G.Don(C.roseus)-AgNPs.METHODS The proliferative activity of hepatocellular carcinoma(HepG2)and normal human liver(THLE3)cells treated with C.roseusAgNPs were measured using MTT assay.The RNA samples were extracted and sequenced using BGIseq500 platform.This is followed by data filtering,mapping,gene expression analysis,differentially expression genes analysis,Gene Ontology analysis,and pathway analysis.RESULTS The mean IC 50 values of C.roseusAgNPs on HepG2 was 4.38±1.59μg/mL while on THLE3 cells was 800±1.55μg/mL.Transcriptome profiling revealed an alteration of 296 genes.C.roseusAgNPs induced the expression of stress-associated genes such as MT,HSP and HMOX-1.Cellular signalling pathways were potentially activated through MAPK,TNF and TGF pathways that are responsible for apoptosis and cell cycle arrest.The alteration of ARF6,EHD2,FGFR3,RhoA,EEA1,VPS28,VPS25,and TSG101 indicated the uptake of C.roseus-AgNPs via both clathrin-dependent and clathrinindependent endocytosis.CONCLUSION This study provides new insights into gene expression study of biosynthesised AgNPs on cancer cells.The cytotoxicity effect is mediated by the aberrant gene alteration,and more interestingly the unique selective antiproliferative properties indicate the C.roseusAgNPs as an ideal anticancer candidate.

Molecular epidemiology of piliated pneumococcal isolates at a major tertiary hospital in the Klang Valley,Malaysia

Objective:To characterise a collection of pili-carrying and none pili-carrying pneumococcal isolates of clinical origin for serotypes,antibiotic resistance and genotype.Methods:In total,42 clinical isolates were collected between October 2017 and December 2019.Those isolates were analysed for antimicrobial susceptibility,serotype distribution,detection of pneumococcal virulence and pilus genes.Multilocus sequence typing was performed only for piliated isolates,followed by phylogenetic analysis.Results:The common isolation sites among the pneumococcal isolates were tracheal aspirate(28.6%),blood(26.2%),and sputum(23.8%).Fifty percent isolates were resistant to erythromycin,tetracycline(50.0%)and trimethoprim-sulfamethoxazole(43.0%).The most frequent were serotypes 19F(28.6%),6A/B(23.8%)and 19A(14.3%).Piliated isolates were detected in a small proportion(33.3%);64.3%were multidrug-resistant.ST320 was the prevalent sequence type among the piliated isolates and genetically related to the Pneumococcal Molecular Epidemiology Network clones Taiwan19F-14(CC271).In the phylogenetic analysis,some piliated isolates showed a close association having similar ST320,carrying serotype 19A and both pilus genes indicating their clonal spread.Conclusions:Pneumococcal lineages of piliated isolates have been globally disseminated and pili could have played a role in the spread of antibiotic resistant clones.

Calf Posture Recognition Using Convolutional Neural Network

Dairy farm management is crucial to maintain the longevity of the farm,and poor dairy youngstock or calf management could lead to gradually deteriorating calf health,which often causes premature death.This was found to be the most neglected part among the management workflows in Malaysia and has caused continuous loss over the recent years.Calf posture recognition is one of the effective methods to monitor calf behaviour and health state,which can be achieved by monitoring the calf behaviours of standing and lying where the former depicts active calf,and the latter,passive calf.Calf posture recognition module is an important component of some automated calf monitoring systems,as the system requires the calf to be in a standing posture before proceeding to the next stage of monitoring,or at the very least,to monitor the activeness of the calves.Calf posture such as standing or resting can easily be distinguished by human eye,however,to be recognized by a machine,it will require more complicated frameworks,particularly one that involves a deep learning neural networks model.Large number of highquality images are required to train a deep learning model for such tasks.In this paper,multiple ConvolutionalNeuralNetwork(CNN)architectures were compared,and the residual network(ResNet)model(specifically,ResNet-50)was ultimately chosen due to its simplicity,great performance,and decent inference time.Two ResNet-50 models having the exact same architecture and configuration have been trained on two different image datasets respectively sourced by separate cameras placed at different angle.There were two camera placements to use for comparison because camera placements can significantly impact the quality of the images,which is highly correlated to the deep learning model performance.After model training,the performance for both CNN models were 99.7%and 99.99%accuracies,respectively,and is adequate for a real-time calf monitoring system.