This research paper aimed to locate protein modifications caused by treatment of milk and determine if the modification locations were consistent. The majority of milk for consumption is homogenised using pressure and heat, and this causes changes in the location of proteins in the milk as well as protein modifications. To investigate these proteomic changes, raw milk was pasteurised (72°C, 15 s), then, to separate the treatment for homogenisation, heated at these different pressures and temperatures: 45°C without no pressure applied, 45°C with 35 MPa, 80°C without pressure applied and 80°C, with 35 MPa. Proteomic analysis was done after separating the milk into three fractions: whey, casein and cream. Protein modifications in each fraction were examined and we found Maillard products as well as oxidation to be of interest. The proteins were also further identified and characterised to compare protein modification sites and differences in proteins present in the cream resulting from homogenisation and/or pasteurisation. This experiment showed that both heat and pressure during homogenisation can cause increases in protein modifications as a result of oxidation or the Maillard reaction. Total cysteine oxidation and total proline oxidation differed between treatments although this was only significantly different for cysteine. It was observed that protein modifications occurred in the same location in the protein sequence rather than in random locations which we highlighted by examining α-S1-casein, lactadherin and β-lactoglobulin.

Phenomenological models are popular for describing the epidemic curve. We present how they can be used at different phases in the epidemic, by modelling the daily number of new hospitalisations (or cases). As real-time prediction of the hospital capacity is important, a joint model of the new hospitalisations, number of patients in hospital and in intensive care unit (ICU) is proposed. This model allows estimation of the length of stay in hospital and ICU, even if no (or limited) individual level information on length of stay is available. Estimation is done in a Bayesian framework. In this framework, real-time alarms, defined as the probability of exceeding hospital capacity, can be easily derived. The methods are illustrated using data from the COVID-19 pandemic in March–June 2020 in Belgium, but are widely applicable.

Background: Pertussis continues to be an important health issue in Vietnam despite infant vaccination programs. In Vietnam, the incidence rates of pertussis per 100,000 population rose from 0.09 in 2014 to 0.33 in 2015 and to 0.58 in 2017. Macrolides, especially erythromycin, are the treatment of choice. However, erythromycin-resistant cases, caused by transition at A2047G position in 23S rRNA, have been reported in the region. Few data are available on antimicrobial resistance in Bordetella pertussis to guide treatment in Vietnam. We report antimicrobial susceptibility of the circulating strains in southern Vietnam during 2015–2017. Methods: Tracheal aspirates from 263 suspected pertussis cases were subject to multiplex real-time PCR to identify B. pertussis and Bordetella spp. Samples were cultured on Regan Lowe agar with 10% sheep blood containing cephalexin (40 µg/mL) and incubated at 37°C for 10 days. The antimicrobial susceptibilities to erythromycin, azithromycin, clarithromycin, and trimethoprim/sulfamethoxazole were determined using the disc diffusion method (CLSI-2017) on Regan Lowe and Mueller Hinton agar. Erythromycin minimum inhibitory concentrations (MICs) were determined using an E-test. The results were recorded after days 3 and 7 of incubation. Sequencing of the 23S rRNA gene was performed to detect mutations conferring macrolide resistance. Results: Of 263 cases, 119 were positive for B. pertussis (45.2%) by real-time PCR, and 15 of 263 strains (5.7%) were successfully cultured. All 15 isolates were susceptible to macrolides and no heterogeneous phenotype was recorded after 7 days; erythromycin MICs were ≤0.094 µg/mL (Fig. 1). We observed no difference in results generated on Regan Lowe and Mueller Hinton media. However, for testing trimethoprim/sulfamethoxazole, results on were superior, as those on Regan Lowe media were unclear. Sequencing of 23S rRNA identified no mutations known to confer macrolide resistance. Conclusions: None of 15 B. pertussis isolates tested were nonsusceptible to erythromycin and macrolides. Similarly, no mutation at the erythromycin-binding site in the 23S rRNA gene was identified. The low isolation rate of B. pertussis by culture means that few positive specimens were tested for antimicrobial susceptibility. To overcome this limitation, detection of resistance directly from clinical specimens needs to be investigated. Ongoing screening for B. pertussis and antimicrobial susceptibility is recommended to support efforts to control the spread of this respiratory tract infection agent.

Funding: None

Disclosures: None

Background: Whooping cough is a serious respiratory illness in infants caused by Bordetella pertussis. In spite of the vaccination program, the incidence rates of whooping cough per 100,000 population in Vietnam increased from 0.33 in 2015 to 0.58 in 2017. If this represents a pertussis resurgence, contributors may include pathogen adaptation, the spread of specific variants, vaccine failure, and failure to effectively treat cases and contacts. There has been little research in Vietnam on B. pertussis strains. Therefore, we investigated the molecular epidemiology of circulating B. pertussis strains in Southern Vietnam by applying multilocus sequence typing (MLST) for 7 housekeeping genes and 4 antigenic determinant genes as components in the acellular vaccine including prn, ptxP, ptxS1, and fim3. Methods: DNA was extracted from 15 isolates collected from 263 case patients during 2015–2017 and was subject to MLST using primers and cycling conditions from the Bordetella pubMLST website (www.Pubmlst.org/Bordetella/). The products were analyzed using BioEdit version 7.2.5 software and then were aligned and compared to reference sequences of each genotype in the database. The evolutionary relationship among sequence types (STs) from housekeeping genes was performed as a minimum spanning tree via the goeBURST algorithm, whereas the correlation of different variants from 4 antigenic determinant genes was built up and clarified with phylogenetic trees based on the UPGMA method by MEGA 7 program. Results: The 15 isolates were all classified as ST2 (100%) by MLST of housekeeping genes, and they belonged to a common global clonal group (Fig. 1). Sequencing of antigenic determinant genes prn2 – ptxP3 – ptxS1-1 – fim3-1 determined that all were identical to each another and the reference sequences (Fig. 2). Conclusions:B. pertussis isolates circulating in Southern Vietnam appeared to be the same as the common global strain. Few isolates were available for testing; therefore, continued surveillance is important to confirm these findings and to monitor population changes over time.

Funding: None

Disclosures: None

To reduce competition with human-edible feed resources, it is of interest to incorporate by-products from the food industry in animal feeds. The current research investigated the effect of including increasing amounts of tofu by-product (TF) in practical pig diets on animal performance, nitrogen balance and ammonia emissions from manure. Two experiments were conducted including a control diet without TF, containing 160 g/kg dietary non-starch polysaccharides (NSPs) and three diets including 122, 246 and 360 g TF/kg DM (TF122, TF246 and TF360, respectively) to reach 220, 280 and 360 g/kg NSP. All diets had the same level of CP and protein digestible in the small intestine which particularly was realized by replacing rice bran with TF. Animal performance was assessed in a first experiment with 40 growing barrows with initial BW of 26.6 ± 1.80 kg (M ± SD) being allocated to the 4 treatments, during 2 growth phases (i.e. until 50 kg BW and from 50 to 80 kg BW). In the growth phase until 50 kg, feed intake and average daily gain (ADG) were linearly reduced by dietary TF inclusion, while this negative impact disappeared during the second growth phase (50 to 80 kg BW). Tofu by-product inclusion even positively affected the feed conversion ratio during this second growth phase (3.4 to 2.7 kg feed/kg ADG for 0 to 360 g/kg dietary TF). Over the entire growth period, performance and feed intake were negatively affected at the highest dietary TF level. Experiment 2 was conducted to assess digestibility, nitrogen balance and ammonia emission from manure. For this purpose, 16 pigs with BW of 62.8 ± 3.6 kg (M ± SD) were assigned to either 1 of the 4 treatments. There was no difference in total tract apparent digestibility of dietary organic matter or CP, while NDF digestibility increased with increasing TF level, suggesting increasing importance of the hindgut fermentation when digesting diets with increasing TF levels. Nevertheless, this was not reflected in increasing levels of faecal volatile fatty acids or purines, nor in reduced manure pH. As a result, ammonia emission from slurry was not reduced through dietary TF inclusion, despite the linear decrease in urinary nitrogen. In conclusion, TF can be included in pigs’ diets up to an inclusion rate of 25% without risk of impaired animal performance; however, this dietary strategy fails to mitigate ammonia emission from slurry.

Gac is a dioecious tropical and perennial climber. The fruit is a rich source of carotenoids and is used in traditional cuisine and medicine. Improving propagation methods using simple techniques would increase production and improve conservation in regional areas. This study evaluated temperature requirements for seed germination, the use of rooting hormones to strike female cuttings and the grafting of female scions onto seedling rootstock. Seed germination was optimised between 25 and 35 °C, with a maximum germination percentage of 91% at 30 °C. However, increasing storage time from 6 to 18 months under laboratory conditions (21 ± 1°C and 60% relative humidity) reduced germination and this was associated with seed weight loss, highlighting the need to develop storage guidelines, particularly for the higher temperature and humidity conditions where Gac is grown. Survival of softwood cuttings was improved from 53 to 77% with indole-3-butyric acid (IBA) (3–5 g/L) and semi-hardwood cuttings did not require IBA treatment. Both splice and wedge grafting techniques achieved a survival rate > 53% and with the youngest rootstock (4 and 8 weeks) this increased to > 85%. Further work could investigate the production potential of crops using cuttings and grafted plants.

Fasciola jacksoni is a significant contributor to the health and mortality of Asian elephants, particularly those in Sri Lanka. Despite the impact of fascioliasis on elephant populations, it is a neglected veterinary disease with limited taxonomic understanding. Molecular characterization and phylogenetic analysis of F. jacksoni were carried out to evaluate its suggested basal position in the Fasciolidae. Adult worms were collected during post-mortem of elephants, and eggs were collected from living elephants in National parks across Sri Lanka. Using the mitochondrial genes nicotinamide dehydrogenase subunit 1 (nad1) and cytochrome oxidase subunit 1 (cox1), and a partial 28S ribosomal DNA (28S rDNA), DNA sequences were generated from the F. jacksoni adult and egg material. Maximum likelihood (ML) phylogenetic analyses did not resolve F. jacksoni to be basal to the Fasciolidae. Furthermore, the ML analyses showed that the genus Fasciola was not monophyletic and that F. jacksoni was a sister species to the deer liver fluke Fascioloides magna. A clear framework is required to determine the taxonomic status of F. jacksoni and this current study provides the first detailed application of molecular techniques from multiple hosts across Sri Lanka with the production of reference DNA sequences for this important parasite.

The Lung Cam expanded stratigraphic succession in Vietnam is correlated herein to the Meishan D section in China, the GSSP for the Permian–Triassic boundary. The first appearance datum of the conodont Hindeodus parvus at Meishan defines the Permian–Triassic boundary, and using published graphic correlation, the Permian–Triassic boundary level has been projected into the Lung Cam section. Using time-series analysis of magnetic susceptibility (χ) data, it is determined that H. parvus arrived at Lung Cam ∼18 kyr before the Permian–Triassic boundary. Data indicate that the Lung Cam section is expanded by ∼90 % relative to the GSSP section at Meishan. Given the expanded Lung Cam section, it is possible to resolve the timing of significant events during the Permian–Triassic transition with high precision. These events include major stepped extinctions, beginning at ∼135 kyr and ending at ∼110 kyr below the Permian–Triassic boundary, with a duration of ∼25 kyr, followed by deposition of Lung Cam ash Bed + 13, which is equivalent to Siberian Traps volcanism is graphically correlated to a precession Time-series model, placing onset of this major volcanic event at ~242 kyr before the PTB. The Meishan Beds 25 and 26, at ∼100 kyr before the Permian–Triassic boundary. In addition, the elemental geochemical, carbon and oxygen isotope stratigraphy, and magnetostratigraphy susceptibility datasets from Lung Cam allow good correlation to other Permian–Triassic boundary succession. These datasets are helpful when the conodont biostratigraphy is poorly known in sections with problems such as lithofacies variability, or is undefined, owing possibly to lithofacies exclusions, anoxia or for other reasons. The Lung Pu Permian–Triassic boundary section, ∼45 km from Lung Cam, is used to test these problems.

A cluster of Salmonella Paratyphi B variant L(+) tartrate(+) infections with indistinguishable pulsed-field gel electrophoresis patterns was detected in October 2015. Interviews initially identified nut butters, kale, kombucha, chia seeds and nutrition bars as common exposures. Epidemiologic, environmental and traceback investigations were conducted. Thirteen ill people infected with the outbreak strain were identified in 10 states with illness onset during 18 July–22 November 2015. Eight of 10 (80%) ill people reported eating Brand A raw sprouted nut butters. Brand A conducted a voluntary recall. Raw sprouted nut butters are a novel outbreak vehicle, though contaminated raw nuts, nut butters and sprouted seeds have all caused outbreaks previously. Firms producing raw sprouted products, including nut butters, should consider a kill step to reduce the risk of contamination. People at greater risk for foodborne illness may wish to consider avoiding raw products containing raw sprouted ingredients.

Background: Outcomes for patients with melanoma brain metastases (MBM) have been poor. New radiotherapy technologies and systemic agents have improved outcomes. Outcomes have rarely been studied at the population-level. We undertook a population-based study investigating changes in management and outcome for patients with MBM in Ontario from 2007-2016. Methods: This was a retrospective population-based cohort study of patients treated for MBM in Ontario from 2007-2016. Melanoma was identified through the Ontario Cancer Registry. Treatments and outcome were described by era (2007-2009, 2010-2012, 2013-2016). Treatment with cranial radiotherapy and drugs were defined using Cancer Care Ontario data and supplemented by physician billing and drug reimbursement data. Neurosurgery was identified using CIHI hospital records. Time to event was investigated using Kaplan-Meier curves. Results: From January 2007-June 2016, 1096 patients with MBM were treated. Whole brain radiation therapy was the first brain-directed treatment in 75.5% of patients in 2007-2009, dropping to 52.0% for 2013-2016. Patients receiving stereotactic radiation or other conformal techniques as the first brain treatment increased from 3.4% in 2007-2009 to 21.3% 2013-2016. Use of BRAF/MEK inhibitors and immunotherapy increased: <2.0% in 2007-2009 to 40.9% 2013-2016. One-year and two-year overall survival (OS) following first brain-directed treatment was greater in 2013-2016: 21.8% at one year and 13.8% after two years (Wilcoxon p=0.001). This compared to 12.3% and 6.4% 2007-2009, and 10.7% and 5.5% 2010-2012. Conclusion: The advent of new radiation technologies and systemic treatments for MBM was associated with increased survival and greater avoidance of whole brain radiotherapy.

We evaluated the impact of the Epic antimicrobial stewardship module (EAM) on the number of interventions, antimicrobial usage, and clinical outcomes. Use of the EAM allowed us to significantly increase the number of ASP antimicrobial reviews and interventions while maintaining a sustained impact on antimicrobial utilization.

When a drop impacts on a deep pool at moderate velocity it forms a hemispheric crater which subsequently rebounds to the original free-surface level, often forming Worthington jets, which rise vertically out of the crater centre. Under certain impact conditions the crater collapse forms a dimple at its bottom, which pinches off a bubble and is also known to be associated with the formation of a very fast thin jet. Herein we use two ultra-high-speed video cameras to observe simultaneously the dimple collapse and the speed of the resulting jet. The fastest fine jets are observed at speeds of approximately $50~\text{m}~\text{s}^{-1}$ and emerge when the dimple forms a cylinder which retracts without pinching off a bubble. We also identify what appears to be micro-bubbles at the bottom of this cylinder, which we propose are caused by local cavitation from extensional stress in the flow entering the jet. The radial collapse of the dimple does not follow capillary-inertial power laws nor is its bottom driven by a curvature singularity, as has been proposed in some earlier studies. The fastest jets are produced by pure inertial focusing and emerge at finite dimple size, bypassing the pinch-off singularity. These jets emerge from the liquid contained originally in the drop. Finally, we measure directly the compression of the central bubble following the pinch-off and the subsequent large volume oscillation, which occurs at frequencies slightly above the audible range at approximately 23 kHz.

We examined the association between a history of smallpox vaccination and immune activation (IA) in a population of antiretroviral therapy-naïve people living with HIV (PLHIV). A cross-sectional study was conducted in Senegal from July 2015 to March 2017. Smallpox vaccination was ascertained by the presence of smallpox vaccine scar and IA by the plasma level of β-2-microglobulin (β2m). The association was analysed using logistic regression and linear regression models. The study population comprised 101 PLHIV born before 1980 with a median age of 47 years (interquartile range (IQR) = 42–55); 57·4% were women. Smallpox vaccine scar was present in 65·3% and the median β2m level was 2·59 mg/l (IQR = 2·06–3·86). After adjustment, the presence of smallpox vaccine scar was not associated with a β2m level ⩾2·59 mg/l (adjusted odds ratio 0·94; 95% confidence interval 0·32–2·77). This result was confirmed by the linear regression model. Our study does not find any association between the presence of smallpox vaccine scar and the β2m level and does not support any association between a previous smallpox vaccination and HIV disease progression. In this study, IA is not a significant determinant of the reported non-targeted effect of smallpox vaccination in PLHIV.

Central nervous system infections (CNSI) are a leading cause of death and long-term disability in children. Using ICD-10 data from 2005 to 2015 from three central hospitals in Ho Chi Minh City (HCMC), Vietnam, we exploited generalized additive mixed models (GAMM) to examine the spatial-temporal distribution and spatial and climatic risk factors of paediatric CNSI, excluding tuberculous meningitis, in this setting. From 2005 to 2015, there were 9469 cases of paediatric CNSI; 33% were ⩽1 year old at admission and were mainly diagnosed with presumed bacterial CNSI (BI) (79%), the remainder were >1 year old and mainly diagnosed with presumed non-bacterial CNSI (non-BI) (59%). The urban districts of HCMC in proximity to the hospitals as well as some outer districts had the highest incidences of BI and non-BI; BI incidence was higher in the dry season. Monthly BI incidence exhibited a significant decreasing trend over the study. Both BI and non-BI were significantly associated with lags in monthly average temperature, rainfall, and river water level. Our findings add new insights into this important group of infections in Vietnam, and highlight where resources for the prevention and control of paediatric CNSI should be allocated.

The identities of two novel perennial nutsedge biotypes collected near Bakersfield, CA, were assessed using isozyme and random-amplified polymorphic deoxyribonucleic acid markers in conjunction with morphological analysis. The two biotypes, designated as CK (Cyperus rotundus cv. ‘Kempeni’) and CR (Cyperus esculentus cv. ‘Robusta’), morphologically resemble purple nutsedge and yellow nutsedge, respectively. Plants from both biotypes exhibited more prolific growth than the typical forms and possessed some traits that are not characteristic of the species they resemble. The morphological study was conducted on a total of 15 purple nutsedge, yellow nutsedge, CK, and CR populations collected in the first year and on 20 additional nutsedge populations were collected in the second year. The genetic analysis was performed on populations from the first year only. In general, there was agreement among the results obtained from the morphometric, isozymatic, and deoxyribonucleic acid studies. Populations of CR clustered with yellow nutsedge populations, indicating that CR is within the normal range of variation of this species and may represent a new introduction. Populations of CK, however, were distinct from both purple nutsedge and yellow nutsedge populations. Considering the low level of genetic variation reported in purple nutsedge and its strict vegetative mode of reproduction, CK might represent a sexually reproducing ecotype of purple nutsedge or a hybrid with yellow nutsedge.

The steady, axisymmetric flow induced by a point sink (or source) submerged in an inviscid fluid of infinite depth is computed and the resulting deformation of the free surface is obtained. The effect of surface tension on the free surface is determined and is the new component of this work. The maximum Froude numbers at which steady solutions exist are computed. It is found that the determining factor in reaching the critical flow changes as more surface tension is included. If there is zero or a very small amount of surface tension, the limiting factor appears to be the formation of small wavelets on the free surface; but, as the surface tension increases, this is replaced by a tendency for the lowest point on the free surface to descend sharply as the Froude number is increased.