Isolation of kistomin milk coagulant from calloselasma rhodostoma venom

Rennet is an enzyme used in the milk clotting process for cheese production. Rennet is produced in the stomach of ruminant mammal especially calf. However, increasing demand for cheese with ongoing research for innovative cheese products has encouraged the researchers to explore the other new sources of milk clotting protease. The new sources of milk clotting protease include animal, plant, microbes and fungi have been investigated. Snake venom contains a rich and good source of protease that needs to be explored as well for its potential in milk clotting. The objectives of this study were to screen eleven different species of snake venoms, to isolate the protease enzyme from the most potential venom capable to coagulate milk using chromatography techniques, to determine the enzymes milk clotting parameters and compare with a commercial milk clotting protease. The results revealed that a crude venom from Calloselasma rhodostoma showed the fastest clotting time that is 2.83±0.17 minutes and was chosen for further purification process. The milk clotting enzyme was purified by HiTrap SP FF ionexchange chromatography and further separated using HiPrep 26/60 Sephacryl S 200 HR size exclusion column. It was purified 4.41±0.06 fold and exhibited recovery activity of 54.34 %. SDS-PAGE analysis indicated a single band with a molecular mass of approximately 26 kDa. The purified protease was completely inhibited by EDTA and 1,10 phenanthroline revealing to be a metalloprotease SVMPs P-I Kistomin confirmed by Mass Spectrometry analysis. The specific activity of kistomin is 62.20 (U/mg). The milk clotting activity (MCA) of kistomin under optimum conditions, was 810.44±42.45 (SU/mL) and the PA of kistomin was 1.39±0.01 (U/mL), resulted in high ratio of MCA/PA value of 585.05. The clotting activity of kistomin on milk was the highest at 0.76 mg/mL kistomin concentration, 8 % (w/v) of calcium chloride concentration, temperature of 48 °C and stable over wide range of pH 5-7 with the peak of pH 6.5. The addition of Ba2+, Mn2+and Ca2+ions as cofactor significantly increased the enzyme activity but inhibited by Hg2+, Pb2+and Fe2+ions. The Km value of kistomin on casein is 1.153±0.08 mg/mL. The low Km value of kistomin on casein showed it has high affinity to casein. Kistomin promoted extensive cleavage of kappa casein and low level of beta casein hydrolysis. From this preliminary study, it can be concluded that kistomin has potential to be a coagulant in the dairy industry. However, due to the source of the protease being from a snake and always been considered lethal by people in general, a thorough safety investigation warranted before been utilised in any industry

Antimicrobial resistance among migrants in Europe: a systematic review and meta-analysis

BACKGROUND: Rates of antimicrobial resistance (AMR) are rising globally and there is concern that increased migration is contributing to the burden of antibiotic resistance in Europe. However, the effect of migration on the burden of AMR in Europe has not yet been comprehensively examined. Therefore, we did a systematic review and meta-analysis to identify and synthesise data for AMR carriage or infection in migrants to Europe to examine differences in patterns of AMR across migrant groups and in different settings. METHODS: For this systematic review and meta-analysis, we searched MEDLINE, Embase, PubMed, and Scopus with no language restrictions from Jan 1, 2000, to Jan 18, 2017, for primary data from observational studies reporting antibacterial resistance in common bacterial pathogens among migrants to 21 European Union-15 and European Economic Area countries. To be eligible for inclusion, studies had to report data on carriage or infection with laboratory-confirmed antibiotic-resistant organisms in migrant populations. We extracted data from eligible studies and assessed quality using piloted, standardised forms. We did not examine drug resistance in tuberculosis and excluded articles solely reporting on this parameter. We also excluded articles in which migrant status was determined by ethnicity, country of birth of participants' parents, or was not defined, and articles in which data were not disaggregated by migrant status. Outcomes were carriage of or infection with antibiotic-resistant organisms. We used random-effects models to calculate the pooled prevalence of each outcome. The study protocol is registered with PROSPERO, number CRD42016043681. FINDINGS: We identified 2274 articles, of which 23 observational studies reporting on antibiotic resistance in 2319 migrants were included. The pooled prevalence of any AMR carriage or AMR infection in migrants was 25·4% (95% CI 19·1-31·8; I2 =98%), including meticillin-resistant Staphylococcus aureus (7·8%, 4·8-10·7; I2 =92%) and antibiotic-resistant Gram-negative bacteria (27·2%, 17·6-36·8; I2 =94%). The pooled prevalence of any AMR carriage or infection was higher in refugees and asylum seekers (33·0%, 18·3-47·6; I2 =98%) than in other migrant groups (6·6%, 1·8-11·3; I2 =92%). The pooled prevalence of antibiotic-resistant organisms was slightly higher in high-migrant community settings (33·1%, 11·1-55·1; I2 =96%) than in migrants in hospitals (24·3%, 16·1-32·6; I2 =98%). We did not find evidence of high rates of transmission of AMR from migrant to host populations. INTERPRETATION: Migrants are exposed to conditions favouring the emergence of drug resistance during transit and in host countries in Europe. Increased antibiotic resistance among refugees and asylum seekers and in high-migrant community settings (such as refugee camps and detention facilities) highlights the need for improved living conditions, access to health care, and initiatives to facilitate detection of and appropriate high-quality treatment for antibiotic-resistant infections during transit and in host countries. Protocols for the prevention and control of infection and for antibiotic surveillance need to be integrated in all aspects of health care, which should be accessible for all migrant groups, and should target determinants of AMR before, during, and after migration. FUNDING: UK National Institute for Health Research Imperial Biomedical Research Centre, Imperial College Healthcare Charity, the Wellcome Trust, and UK National Institute for Health Research Health Protection Research Unit in Healthcare-associated Infections and Antimictobial Resistance at Imperial College London

Surgical site infection after gastrointestinal surgery in high-income, middle-income, and low-income countries: a prospective, international, multicentre cohort study

Background: Surgical site infection (SSI) is one of the most common infections associated with health care, but its importance as a global health priority is not fully understood. We quantified the burden of SSI after gastrointestinal surgery in countries in all parts of the world. Methods: This international, prospective, multicentre cohort study included consecutive patients undergoing elective or emergency gastrointestinal resection within 2-week time periods at any health-care facility in any country. Countries with participating centres were stratified into high-income, middle-income, and low-income groups according to the UN's Human Development Index (HDI). Data variables from the GlobalSurg 1 study and other studies that have been found to affect the likelihood of SSI were entered into risk adjustment models. The primary outcome measure was the 30-day SSI incidence (defined by US Centers for Disease Control and Prevention criteria for superficial and deep incisional SSI). Relationships with explanatory variables were examined using Bayesian multilevel logistic regression models. This trial is registered with ClinicalTrials.gov, number NCT02662231. Findings: Between Jan 4, 2016, and July 31, 2016, 13 265 records were submitted for analysis. 12 539 patients from 343 hospitals in 66 countries were included. 7339 (58·5%) patient were from high-HDI countries (193 hospitals in 30 countries), 3918 (31·2%) patients were from middle-HDI countries (82 hospitals in 18 countries), and 1282 (10·2%) patients were from low-HDI countries (68 hospitals in 18 countries). In total, 1538 (12·3%) patients had SSI within 30 days of surgery. The incidence of SSI varied between countries with high (691 [9·4%] of 7339 patients), middle (549 [14·0%] of 3918 patients), and low (298 [23·2%] of 1282) HDI (p < 0·001). The highest SSI incidence in each HDI group was after dirty surgery (102 [17·8%] of 574 patients in high-HDI countries; 74 [31·4%] of 236 patients in middle-HDI countries; 72 [39·8%] of 181 patients in low-HDI countries). Following risk factor adjustment, patients in low-HDI countries were at greatest risk of SSI (adjusted odds ratio 1·60, 95% credible interval 1·05–2·37; p=0·030). 132 (21·6%) of 610 patients with an SSI and a microbiology culture result had an infection that was resistant to the prophylactic antibiotic used. Resistant infections were detected in 49 (16·6%) of 295 patients in high-HDI countries, in 37 (19·8%) of 187 patients in middle-HDI countries, and in 46 (35·9%) of 128 patients in low-HDI countries (p < 0·001). Interpretation: Countries with a low HDI carry a disproportionately greater burden of SSI than countries with a middle or high HDI and might have higher rates of antibiotic resistance. In view of WHO recommendations on SSI prevention that highlight the absence of high-quality interventional research, urgent, pragmatic, randomised trials based in LMICs are needed to assess measures aiming to reduce this preventable complication

Mahasiswa UMP hulur bantuan kanak-kanak mangsa taufan Haiyan dan gelandangan

Cebu (Filipina), 22 Ogos – Pengalaman 10 mahasiswa Universiti Malaysia Pahang (UMP) bersama-sama pihak badan kebajikan Mercy in Action yang merupakan badan kebajikan swasta yang memfokuskan kanak-kanak dan komuniti setempat pastinya meninggalkan pengalaman yang penuh makna buat mereka yang berpeluang mengikuti Program Jelajah Komuniti ASEAN 2015: “Komuniti Sosial & Budaya Filipina” anjuran Jabatan Hal Ehwal Pelajar dan Alumni (JHEPA) baru-baru ini

Mahasiswa hulur bantuan kanak-kanak mangsa taufan Haiyan dan gelandangan di Filipina

Pengalaman 10 mahasiswa Universiti Malaysia Pahang (UMP) bersama-sama pihak badan kebajikan Mercy in Action yang merupakan badan kebajikan swasta yang memfokuskan kanak-kanak dan komuniti setempat pastinya meninggalkan pengalaman yang penuh makna buat mereka yang berpeluang mengikuti Program Jelajah Komuniti ASEAN 2015: 'Komuniti Sosial & Budaya Filipina' anjuran Jabatan Hal Ehwal Pelajar dan Alumni (JHEPA) pada 22 Ogos 2015 yang lalu

Enzymic Milk Clotting Activities of Metalloproteinase Kistomin

Background and Objectives: The dairy industry is in constant search for newer milk clotting coagulants to substitute rennet due to its growing demands and controversies such as; its origin from young slaughtered animals. Kistomin (EC:3.4.24) is a protease long identified on its role to cleave platelet and fibrinogen. In this study, kistomin discovered to function as a milk coagulant and investigated for its milk clotting activities. Materials and Methods: Kistomin investigated by measuring its Milk Clotting Activity (MCA) and Proteolytic Activity (PA), optimum conditions of pH, temperature, concentrations of enzyme and calcium chloride, exposures to various chelating agents and cofactor ions. Results: The MCA of kistomin was 810.44 (SU mLG1 ) and the PA was 1.39 (U mLG1 ), resulted in a ratio of MCA/PA value of 583. The coagulating activity of kistomin on milk was the highest at 0.76 mg mLG1 enzyme concentration, 8% (w/v) of CaCl2 concentration, the temperature of 48EC and stable over a wide range of pH 5-7 with activity peaking at pH6.5. The protease completely inhibited by EDTA and 1,10 phenanthroline verifying to be a metalloprotease. The addition of Ba2+, Mn2+ and Ca2+ significantly increased the enzyme activity but inhibited by Hg2+, Pb2+ and Fe2+ ions. Kistomin promoted extensive hydrolysis of κ-casein and low level of β-casein cleavage. Conclusion: This concluded that kistomin can be used as a milk clotting candidate for the dairy industry

Uncovering a Protease in Snake Venom Capable To Coagulate Milk to Curd

Snake venom been studied for its lethality and various benefits for mankind. The latter been studied a plenty of recent but none related to coagulation of milk to curd. The coagulation time of milk by samples were done using visible parameters i.e. change in viscosity, colour changes, white spot formation (separation between curd and whey) and finally observing a drop of coagulating fluid under magnification of a light microscope. Optimum parameters determined included concentration of coagulants, temperature and pH. Microscopic viewing included observing after centrifugation, under light microscope and SEM. Screening eleven venoms mostly predominantly found in tropical region singled out one with the most rapid coagulating timei.e by Calloselasma rhodostoma (CR). Optimization of CR venom related to several parameters provided venom concentration, 0.07 (w/v%); pH,7.0; temperature, 45.50C while that of rennet were determined to be 0.04±0.02 (w/v%); pH,7.0; temperature, 45.50C, respectively. Under these ideal conditions for both coagulants, comparison of their milk coagulation time found CR superior i.e. 0.41±0.02 min compared to 4.23±0.05 min for rennet. Milk coagulating assay guided fractionation of CR venom by using HiTrap SP FF and consecutively followed by HiPrep 26/60 Sephacryl S200 HR pre-packed columns led to a single band on coomassie stained SDS-PAGE gel. Next by LCMS analysis on the SDS PAGE band identified the presence of metalloproteinase kistominwithin the venom. EDTA inactivated the venom presumably chelating zinc hence suggesting further towards identifying kistomin as the likely protease within this venom with milk-clotting activity. Snake venom been potentially identified for yet another application for the benefit of mankind. In this investigation Malayan Pit Viper’s protease can play major role in dairy industry if studied further

Marker to authenticate Eurycoma longifolia (Tongkat Ali) containing aphrodisiac herbal products

The benefits of Eurycoma longifolia (Tongkat Ali, TA) for its aphrodisiac capabilities are well known and many products are marketed worldwide. Due to its popularity, the plant is being abused for promoting fake products. Therefore, there is a need for better testing of the markers required by authorities and responsible manufacturers. A low-molecular-weight protein has been studied for developing it as a testing marker. Two dimensional electrophoresis (2DE) (four spots were observed) was used for positive detection of proteins in an aqueous extract of TA root and the pronounced separation of a Coomassie-stained spot, subsequently referred to as Marker A. Consecutive chromatographic separations of the aqueous extract of TA led to the isolation of pure protein from Marker A. When this marker was used to test 46 TA-based products randomly selected from markets worldwide, in 20 of them, the results were found to be comparable to those obtained using the organic eurycomanone marker. The ranking of products from the highest quantity to the lowest observed to be ordered differently if compared for both markers. This is an expected outcome because Marker A was measured for its protein content and eurycomanone for its organic molecule. Marker A detection using 2DE is shown to be a useful tool to test products supplemented with E. longifolia root

Surgical site infection after gastrointestinal surgery in children : an international, multicentre, prospective cohort study

Introduction Surgical site infection (SSI) is one of the most common healthcare-associated infections (HAIs). However, there is a lack of data available about SSI in children worldwide, especially from low-income and middle-income countries. This study aimed to estimate the incidence of SSI in children and associations between SSI and morbidity across human development settings. Methods A multicentre, international, prospective, validated cohort study of children aged under 16 years undergoing clean-contaminated, contaminated or dirty gastrointestinal surgery. Any hospital in the world providing paediatric surgery was eligible to contribute data between January and July 2016. The primary outcome was the incidence of SSI by 30 days. Relationships between explanatory variables and SSI were examined using multilevel logistic regression. Countries were stratified into high development, middle development and low development groups using the United Nations Human Development Index (HDI). Results Of 1159 children across 181 hospitals in 51 countries, 523 (45 center dot 1%) children were from high HDI, 397 (34 center dot 2%) from middle HDI and 239 (20 center dot 6%) from low HDI countries. The 30-day SSI rate was 6.3% (33/523) in high HDI, 12 center dot 8% (51/397) in middle HDI and 24 center dot 7% (59/239) in low HDI countries. SSI was associated with higher incidence of 30-day mortality, intervention, organ-space infection and other HAIs, with the highest rates seen in low HDI countries. Median length of stay in patients who had an SSI was longer (7.0 days), compared with 3.0 days in patients who did not have an SSI. Use of laparoscopy was associated with significantly lower SSI rates, even after accounting for HDI. Conclusion The odds of SSI in children is nearly four times greater in low HDI compared with high HDI countries. Policies to reduce SSI should be prioritised as part of the wider global agenda.Peer reviewe

Exploring the cost-effectiveness of high versus low perioperative fraction of inspired oxygen in the prevention of surgical site infections among abdominal surgery patients in three low- and middle-income countries

Background: This study assessed the potential cost-effectiveness of high (80–100%) vs low (21–35%) fraction of inspired oxygen (FiO2) at preventing surgical site infections (SSIs) after abdominal surgery in Nigeria, India, and South Africa. Methods: Decision-analytic models were constructed using best available evidence sourced from unbundled data of an ongoing pilot trial assessing the effectiveness of high FiO2, published literature, and a cost survey in Nigeria, India, and South Africa. Effectiveness was measured as percentage of SSIs at 30 days after surgery, a healthcare perspective was adopted, and costs were reported in US dollars ($). Results: High FiO2 may be cost-effective (cheaper and effective). In Nigeria, the average cost for high FiO2 was$216 compared with $222 for low FiO2 leading to a −$6 (95% confidence interval [CI]: −$13 to −$1) difference in costs. In India, the average cost for high FiO2 was $184 compared with$195 for low FiO2 leading to a −$11 (95$15 to −$6) difference in costs. In South Africa, the average cost for high FiO2 was$1164 compared with $1257 for low FiO2 leading to a −$93 (95% CI: −$132 to −$65) difference in costs. The high FiO2 arm had few SSIs, 7.33% compared with 8.38% for low FiO2, leading to a −1.05 (95% CI: −1.14 to −0.90) percentage point reduction in SSIs. Conclusion: High FiO2 could be cost-effective at preventing SSIs in the three countries but further data from large clinical trials are required to confirm this