Genetic diversity and molecular epidemiology of outbreaks of Klebsiella pneumoniae mastitis on two large Chinese dairy farms

Klebsiella pneumoniae is an opportunistic and environmental mastitis-causing pathogen, with potential for contagious transmission. Repetitive element sequence-based PCR was used to determine genetic diversity and explore potential transmission and reservoirs for mastitis caused by K. pneumoniae on 2 large Chinese dairy farms. A total of 1,354 samples was collected from the 2 dairy farms, including milk samples from cows with subclinical and clinical mastitis, bedding, feces, feed, teat skin, and milking liners. Environmental samples were collected from all barns and milking parlors and extramammary samples from randomly selected dairy cows on both farms. In total, 272 and 93 K. pneumoniae isolates were obtained from Farms A and B, respectively (with ~8K and 2K lactating cows, respectively). Isolation rates from clinical mastitis (CM), subclinical mastitis (SCM), and environmental or extramammary samples were 34, 23 and 37%, respectively for Farm A and 42, 3, and 34% for Farm B. The K. pneumoniae isolated from CM milk and extramammary or environmental sources had high genetic diversity (index of diversity >90%) on the 2 farms and from SCM on Farm A. However, on Farm B, 9 SCM isolates were classified as 2 genotypes, resulting in a relatively low index of diversity (Simpson's index of diversity = 0.39; 95% CI = 0.08–0.70). Genotypes of K. pneumoniae causing mastitis were commonly detected in feces, bedding, and milking liners (Farm A), or from teat skin, sawdust bedding, and feed (Farm B). Based on its high level of genetic diversity, we inferred K. pneumoniae was an opportunistic and environmental pathogen causing outbreaks of CM on these 2 large Chinese dairy farms. Nevertheless, that only a few genotypes caused SCM implied some strains had increased udder adaptability and a contagious nature or a common extramammary source. Finally, control of intramammary infections caused by K. pneumoniae on large Chinese dairy farms must consider farm-level predictors, as the 2 outbreaks had distinct potential environmental sources of infection.     

Composition and biological effects of goldenberry byproducts: an overview

Goldenberry is a wild fruit that has been widely used for centuries, mainly in folk medicine. Most studies of goldenberry have focused on the fruit, but new research has studied its byproducts, which were considered to be waste until recently. The main objective of our study was to systematize the published information regarding the composition of goldenberry byproducts (calyces, leaves, seeds, and pomace) and their effects on biological systems. Goldenberry byproducts contain minerals, amino acids, withanolides, flavonoids, and essential fatty acids, thus representing good sources of these compounds. Some of their major biological effects include anti-inflammatory, antioxidant, antidiabetic, and antiproliferative effects. Information regarding their toxicity is also presented here. To determine the optimal dosage, further safety studies would be recommended to ensure the best health benefits of these compounds. The available evidence has demonstrated the nutritional value of different byproducts of goldenberry, suggesting them to be potential candidates for use in the cosmetic industry, in the preparation of functional foods, and in phytomedicine for the prevention and adjuvant treatment of some diseases. © 2020 Society of Chemical Industry     

Molecular epidemiology and distribution of antimicrobial resistance genes of Staphylococcus species isolated from Chinese dairy cows with clinical mastitis

Staphylococcus species, categorized into Staphylococcus aureus and non-aureus staphylococci (NAS), are frequent causes of mastitis in dairy cattle around the world. Current treatments using antimicrobials are under increasing scrutiny due to rising prevalence of multi-drug resistance in S. aureus. Objectives of this study were to determine: (1) genetic diversity of Staphylococcus species isolated from clinical mastitis in cows from large Chinese dairy farms; and (2) prevalence and distribution of antimicrobial resistance genes (ARG) in these isolates. Staphylococcus aureus (n = 96) were isolated from 26 herds located in 12 provinces of China, whereas NAS (n = 112) were isolated from 59 herds located in 18 provinces of China. The NAS were identified at the species level using a partial 16S rRNA sequencing method, whereas random amplification of polymorphic DNA (RAPD) PCR was done to determine genetic relationships of isolates. Finally, PCR was used to detect resistance and biofilm formation genes. Staphylococcus chromogenes (33%) was the most common NAS species, followed by Staphylococcus sciuri (17%) and Staphylococcus epidermidis (8%). Staphylococcus aureus was grouped in 12 genotypes, of which 2 types represented 56% of isolates. Staphylococcus chromogenes (n = 37) clustered into 8 RAPD types, with 2 prevalent types containing 73% of isolates. The most prevalent ARG in S. aureus isolates was blaZ (95%), followed by tetM (33%), tetK (31%), ermT (26%), and aacA-aphD (23%). The mecA and vanA were detected in 16 and 4% of isolates, respectively. In NAS, blaZ (100%), mecA (73%), tetK (79%), tetM (96%), mphC (63%), and msrA (54%) were frequently detected. Antimicrobial resistance genes mecA, tetK, tetL, tetM, dfrG, ermB, msrA, mphC, aadD, and aphA3 were more commonly detected in NAS than in S. aureus. Biofilm formation genes (icaA and icaD) were frequently detected in staphylococci isolated from bovine clinical mastitis. The existence of predominant RAPD types in S. aureus and S. chromogenes isolates across Chinese dairy farms indicated that specific genotypes had disseminated within herds and become more udder-adapted. High prevalence of ARG, especially in NAS, highlighted the risk of selection of multi-drug resistant staphylococci with potential as a reservoir of ARG.     

Stability of microencapsulated lactic acid bacteria under acidic and bile juice conditions

The probiotic strains Lactobacillus brevis CCMA1284 and Lactobacillus plantarum CCMA0359 were microencapsulated by spray drying using different matrices – whey powder (W), whey powder with inulin (WI) and whey powder with maltodextrin (WM). Viability of the microencapsulated strains in acid and bile juices and during 90 days of storage (seven and 25 °C) was evaluated. The two strains exhibited high encapsulation efficiency (> 86%) by spray drying. The different matrices maintained L. plantarum viability above six log CFU g⁻¹ at 7 °C for 90 days, whereas similar results for L. brevis were observed only for W. The use of inulin as matrix of encapsulation did not enhance bacterial viability in the evaluated conditions. In general, the use of W and WM as matrices was effective for L. plantarum viability. However, only W was effective for L. brevis in the evaluated conditions. The spray drying technique was successfully adopted for the encapsulation of L. plantarum CCMA0359 and L. brevis CCMA1284 strains.     

Resistant starch and thermal,morphological and textural properties of heat‐moisture treated rice starches with high‐, medium‐ and low‐amylose content

This study investigated the effects of heat‐moisture treatment (HMT) on the resistant starch content and thermal, morphological, and textural properties of rice starches with high‐, medium‐ and low‐amylose content. The starches were adjusted to 15, 20 and 25% moisture levels and heated at 110°C for 1 h. The HMT increased the resistant starch content in all of the rice starches. HMT increased the onset temperature and the gelatinisation temperature range (T_finish–T_onset) and decreased the enthalpy of gelatinisation of rice starches with different amylose contents. This reduction increased with the increase in the moisture content of HMT. The morphology of rice starch granules was altered with the HMT; the granules presented more agglomerated surface. The HMT affected the textural parameters of rice starches; the high‐ and low‐amylose rice starches subjected to 15 and 20% HMT possessed higher gel hardness.     

Ball Milling of Amaranth Starch-Enriched Fraction. Changes on Particle Size,Starch Crystallinity,and Functionality as a Function of Milling Energy

Starch-enriched fractions of amaranth grain were obtained from planetary ball milling and subsequently studied for particle size reduction, hydration properties, and crystallinity loss. Wide-angle X-ray scattering (WAXS) was used to evaluate the crystalline of starch-enriched fractions, using an iterative smoothing algorithm to estimate amorphous background scattering. This methodology was then used to determine initial crystallinity and monitor crystallinity loss during this process. The attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR) showed that ball-milling treatment significantly decreased (p?<?0.05) the intensity ratios of the bands at 1,039 and 1,014 cm^?1 corresponding to the crystalline/amorphous part of starch structure. Starch crystallinity degree decreased by ball milling due to starch amorphization during this process. An excellent correlation was found between crystallinity degree obtained by WAXS and ATR-FTIR data for the whole ball-milled-analyzed samples. The energy required for size reduction was satisfactorily explained using a generalized grinding equation. A decrease of span and median diameter (D ₅₀) indicated sample homogenization during ball milling. Water absorption index and water solubility increased with crystallinity loss during process. The flour produced at the higher milling energy (6.52 kJ/g), with a mean size of 68?±?1 μm, showed a low crystallinity degree (<5 %), and high water absorption and solubility indexes in comparison to the starch-enriched fraction sample. Particle activation provided by ball-milling process can offer chances for starch application such as sorbent agent in food or pharmaceutical industries.     

Texture Prediction in Intact Green Asparagus by Near Infrared (NIR) Spectroscopy,Assaying Linear and Non-linear Regression Strategies

Texture-related parameters were assessed in intact green asparagus at harvest and during postharvest storage using near-infrared spectroscopy combined with MPLS and LOCAL algorithms. Three spectrophotometers were evaluated for this purpose: a monochromator (range, 400–2,500 nm), a diode-array Vis–NIR spectrophotometer (range, 400–1,700 nm), and a handheld micro-electro-mechanical system (MEMS) spectrophotometer (range, 1,600–2,400 nm). Three hundred green asparagus spears (cv. “Grande”) were used to obtain calibration models based on reference data and NIR data. Results for maximum shear force showed that LOCAL algorithm improved the predictive capacity of models constructed using all three NIRS instruments, increasing r ² by 24, 16, and 56 % and reducing the SEP(c) values by 11, 8, and 14 %, respectively. For cutting energy, the LOCAL also improved the predictive capacity of the models (r ² increased by 3 % for the monochromator and the diode-array instrument and by 6 % for the MEMS device; and the SEP(c) decreased by 3 % in the three instruments). It is worth noting that while the monochromator and diode-array instruments displayed similar predictive capacity for the parameters tested, the MEMS instrument achieved slightly poorer results but has clear advantages for the measurement of texture in intact asparagus, being economical, portable, and easy to use in situ.     

Optimisation of the extraction of phenolic compounds and antioxidant activity from aerial parts of Bidens pilosa L. using response surface methodology

Response surface methodology was used to find the optimum ethanol concentration and temperature which maximises the antioxidant activity (AA) of hydroalcoholic extracts from aerial parts of Bidens pilosa L. A rotatable central composite design was used, and the extracts were characterised by the determination of solid concentration (SC), total flavonoid (TFC) and total polyphenol content (TPC). AA was determined through 2,2′‐azinobis (3‐ethylbenzothiazoline‐6‐sulphonic acid) diammonium salt (ABTS) and 2,2‐diphenyl‐1‐pycrylhydrazyl (DPPH) radical scavenging activity. Mathematical models showed the significant effects of each variable and allowed to select the optimum conditions of ethanol concentration (62.7%) and extraction temperature (66.2 °C). The optimised extract presented an AA of 804.9 ± 12.2 Trolox equivalent antioxidant capacity (TEAC) dry base (d.b.) for DPPH and 515.8 ± 31.8 TEAC d.b. for ABTS. It was observed that both TFC and TPC showed a good correlation with AA of the extracts.