Methicillin‐Resistant Staphylococcus aureus from Brazilian Dairy Farms and Identification of Novel Sequence Types

The aim of this study was to investigate the phenotypic and genotypic diversity and anti‐microbial resistance among staphylococci of dairy herds that originated from Paraiba State, north‐eastern Brazil, a region where such studies are rare. Milk samples (n = 552) were collected from 15 dairy farms. Isolates were evaluated for anti‐microbial susceptibility by Kirby–Bauer disc diffusion method. Confirmation of methicillin‐resistant Staphylococcus aureus (MRSA) was performed using multiplex PCR targeting mecA and nuc genes in addition to phenotypic assay based on PBP‐2a latex agglutination. Clonal relatedness of isolates was determined by pulsed‐field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) genotyping. Staphylococci were detected in 269 (49%) of the samples. Among these, 65 (24%) were S. aureus. The remaining 204 isolates were either coagulase‐negative staphylococci (n = 188; 70%) or coagulase positive other than S. aureus (n = 16; 6%). Staphylococci were cultured in seven (35%) of the 20 hand swab samples, from which five isolates were S. aureus. The isolates were most commonly resistant against penicillin (43%), ampicillin (38%) and oxacillin (27%). The gene mecA was detected in 21 S. aureus from milk and in one isolate from a milker's hand. None of the isolates were resistant to vancomycin. PFGE findings showed high clonal diversity among the isolates. Based on MLST, we identified a total of 11 different sequence types (STs 1, 5, 6, 83, 97, 126, 1583, 1622, 1623, 1624 and 1625) with four novel STs (ST1622‐ST1625). The findings show that MRSA is prevalent in milk from semi‐extensive dairy cows in north‐eastern Brazil, and further investigation on its extent in various types of milk production systems and the farm‐to‐table continuum is warranted.     

Radiographic assessment of cattle hoofs (ungulae) of the Nellore breed: Reference standards

The purpose of this work was to determine a radiographic standard of normality for lateral and medial ungulae in 3‐year‐old bovine females of the Nellore breed. Twenty‐six cows underwent radiographic exposure of the medial and lateral ungulae of the hindlimbs and forelimbs; abaxioaxial projections were used. The images were analyzed; distances were measured between the dorsal surface of the distal phalanx and the surface of the hoof capsule. The two angles formed, i.e., an angle between the dorsal surface of the distal phalanx and a line parallel to the sole (inner), and an angle between the dorsal surface of the hoof and a line parallel to the sole (outer), were also recorded. The results suggested a standard of normality for the proximal, middle and distal distances of the ungulae of the forelimbs: 0.96 ± 0.12 cm. For the measurement of hindlimbs, except for the distal distances of the side ungulae, the suggested value was 1.02 ± 0.14 cm; for the distal measurement of the lateral ungulae of the hindlimbs, the value was 1.09 ± 0.12 cm. For inner and outer angles, the values were 36.45° ± 4.4° for the medial ungulae of the forelimbs and 42.50° ± 3.8° for other ungulae of the forelimbs and hindlimbs (standard deviations of the values suggested for the angles should be used as variations between different animals, limbs and ungulae; they should not be used between angles of the same ungula).     

Early ovine preantral follicles have a potential to grow until antral stage in two‐step culture system in the presence of aqueous extract of Justicia insularis

The objective of this study was to determine whether preantral follicles cultured in vitro for 7 days within ovine ovarian cortical strips could be isolated at the secondary follicles (SF) and grown until antral stage during an additional 6 days period of in vitro culture in the presence of aqueous extract of Justicia insularis. Fresh ovarian fragments from 16 adult sheep were fixed for histological analysis (Control 1) or in vitro cultured individually in α‐MEM⁺ supplemented with 0.3 mg/ml J. insularis (Step 1) for 7 days. Part of the fragments then were fixed for histological analysis (in vitro culture group). Remaining fragments were exposed stepwise to increasing trehalose concentrations before immediate isolation of SF and viability assessment (Control 2) or after 6 days of culture in α‐MEM⁺⁺ supplemented with 0.3 mg/ml J. insularis (Step 2). In Step 1, percentage of follicular activation was 80%. In Step 2, a significant increase (p < 0.05) in follicular diameter and antrum formation within 6 days in vitro culture of isolated follicles was achieved. The total antioxidant capacity from both steps significantly increase (p < 0.05) from day 2 to day 6. Confocal analysis of oocytes showed 57.14% oocytes with homogeneous distribution and 42.86% with peri‐cortical distribution. In conclusion, SF can be successfully isolated from sheep ovarian cortex after 7 days of culture and are capable of surviving and forming an antral cavity if cultured in vitro for an additional 6 days in the presence of 0.3 mg/ml J. insularis.     

Polymorphism,coupling interval and prematurity index in dogs with degenerative mitral valve disease and ventricular arrhythmias

Ventricular arrhythmias (VA) are a recognized concern in dogs with degenerative mitral valve disease (DMVD). The coupling interval (CI) and the prematurity index (PI) have been shown to accurately differentiate between benign and malignant VA in people, where ventricular arrhythmias are known to be associated with an increased risk of development of signs of heart failure or sudden death. In this study, we characterized ventricular arrhythmias in dogs with symptomatic and asymptomatic DMVD. Seventy dogs with naturally-occurring DMVD and ventricular arrhythmias were retrospectively studied. A cross-sectional investigation including dogs with either symptomatic (stages C/D; n?=?41) or asymptomatic (stages B1/B2; n?=?29) DMVD was performed. Electrocardiographic tracings were reviewed to calculate both the CI and PI. In eight dogs these indices were compared with those obtained from both a Holter recording and a standard ECG tracing and no statistical differences were found (CI, p?=?0.97; PI, p?=?0.17). Even though CI and PI were determined in all animals enrolled in the study, VPC characteristics were only compared between symptomatic and asymptomatic dogs when a 24-h Holter recording was available (n?=?49). The PI was different (p?=?0.01) between symptomatic (0.65?±?0.17) and asymptomatic (0.56?±?0.18) dogs, but CI was considered similar (p?=?0.91). Also, the symptomatic dogs had more polymorphic VPC (p?=?0.002) and supraventricular arrhythmias (p?=?0.0002) than the asymptomatic animals. Polymorphism, and repeating patterns of ventricular premature complexes, were characteristics frequently present in overtly symptomatic animals affected by mitral endocardiosis.     

QT instability,an indicator of augmented arrhythmogenesis,increases with the progression of myxomatous mitral valve disease in dogs

Objectives

To investigate QT instability in dogs with myxomatous mitral valve disease (MMVD) and to determine if this is associated with arrhythmogenesis.

Assessment of longitudinal systolic function using tissue motion annular displacement in healthy dogs

Introduction

Left ventricular systolic function is one of the main parameters studied in echocardiography. Longitudinal systolic function, however, is less commonly evaluated in routine examinations but may provide early information on systolic dysfunction. The movement of the mitral annulus toward the apex has already been determined as a method for evaluation of longitudinal systolic function in dogs, but the study of this movement by speckle tracking with the tissue motion annular displacement (TMAD) technique has not yet been evaluated.

Larvicidal,molluscicidal and nematicidal activities of essential oils and compounds from <Emphasis Type="Italic">Foeniculum vulgare</Emphasis>

Plant-based products, namely essential oils (EOs), are environmentally friendly alternatives for the control of disease vectors, hosts and/or parasites. Here, we studied the general toxicity and biopesticidal potential of EOs and phenylpropanoids from Foeniculum vulgare var. vulgare (bitter fennel), a perennial plant well adapted to temperate climates. EO/compound toxicity was tested against a freshwater snail and potential intermediate host of Fasciola hepatica (Radix peregra), a mosquito and former European malaria vector (Anopheles atroparvus) and one of the most damaging plant-parasitic nematodes, the root-knot nematode (Meloidogyne javanica). Lethal concentrations (LC₅₀; LC₉₀) of EOs (infrutescences/stems with leaves) and compounds were calculated by probit analysis. All displayed noteworthy activity against R. peregra adults (LC₅₀ 21–39 µg ml^?1) and A. atroparvus larvae (LC₅₀ 16–56 µg ml^?1). trans-Anethole revealed acute nematicidal activity after 24 and 48 h (LC₅₀ 310 and 249 µg ml^?1, respectively), and estragole (1,000 µg ml^?1) showed some effectiveness against M. javanica hatching and juveniles after 15 days. Plant and EO yields were determined to evaluate the bitter fennel productivity. The chemical composition of the EOs was analyzed by gas chromatography coupled to mass spectrometry. EOs extracted from whole plants, infrutescences and stems with leaves were characterized by estragole-dominant profiles (28–65 %), considerable amounts of phellandrene (10–34 %) and fenchone (6–16 %), and minor trans-anethole contents (1–4 %). Although additional toxicological studies against nontarget organisms are required, our study demonstrates that bitter fennel is a productive source of molluscicides and larvicides, and thus a potential sustainable biological agent to control particular host species, namely freshwater snails and mosquitoes.     

Practical implications of using non-relational databases to store large genomic data files and novel phenotypes

The objective of our study was to provide practical directions on the storage of genomic information and novel phenotypes (treated here as unstructured data) using a non-relational database. The MongoDB technology was assessed for this purpose, enabling frequent data transactions involving numerous individuals under genetic evaluation. Our study investigated different genomic (Illumina Final Report, PLINK, 0125, FASTQ, and VCF formats) and phenotypic (including media files) information, using both real and simulated datasets. Advantages of our centralized database concept include the sublinear running time for queries after increasing the number of samples/markers exponentially, in addition to the comprehensive management of distinct data formats while searching for specific genomic regions. A comparison of our non-relational and generic solution, with an existing relational approach (developed for tabular data types using 2 bits to store genotypes), showed reduced importing time to handle 50M SNPs (PLINK format) achieved by the relational schema. Our experimental results also reinforce that data conversion is a costly step required to manage genomic data into both relational and non-relational database systems, and therefore, must be carefully treated for large applications.     

Within and between-tree variation in the biometry of wood rays and fibres in cork oak (Quercus suber_L.)

The variability of fibre and ray dimensions was studied at three radial positions (10, 50 and 90% of radius) in 5 cork oaks (Quercus suber L.) approximately 40 years old. Uniseriate ray height and cell number as well as multiseriate ray height and width were measured in tangential sections. Fibre length, width and wall thickness were measured after maceration. The most striking features of cork oak wood were the large multiseriate rays and the frequent homocellular uniseriate rays. On average multiseriate rays were 5.16 mm high and 0.48 mm wide, and uniseriate rays contained 11 cells and were 227 μm high. Neighbour rays presented large dimensional differences but there was no radial variation and there were very small between-tree differences. The radial maintenance of high and wide rays was discussed as a means to improve water shortage in response to water stress. Fibre dimensions increased radially but differed little between trees: on average, 960–1,220 μm length, 18.40–21.49 μm width and 6.66–8.07 μm wall thickness. The ray pattern showed enough variation for visual appreciation while ensuring within and between-tree uniformity. The presence of rays has to be taken into consideration for designing wood drying programs.     

Triterpenic and other lipophilic components from industrial cork byproducts

The detailed chemical composition of the lipophilic extractives of cork and cork byproducts generated throughout industrial processing has been investigated by gas chromatography-mass spectrometry. Triterpenes (cerine, friedeline, and betulinic acid) were the major components detected. Betulinic acid is the main triterpene (11.7 g/kg) identified in industrial cork powder, whereas in black condensates friedeline (95.3 g/kg), betuline (13.1 g/kg), and betulinic acid (12.1 g/kg) are the main triperpenes. Significant fractions of alpha-hydroxy fatty acids (115.1 g/kg) and alpha,omega-dicarboxylic acids (21.2 g/kg) were also detected in black condensate after alkaline hydrolysis. The results demonstrate that these two industrial byproducts can be considered as promising sources of bioactive chemicals or chemical intermediates for the synthesis of polymeric materials.