Prevalence of methicillin-resistant Staphylococci on a farm: staff can harbour MRS when animals do not

The aim of this work was to establish the prevalence of methicillin-resistant Staphylococci (MRS) in the animals and staff of a teaching and research farm. Samples of dairy cattle (36), beef cattle (26), sheep (19), horses (21), pigs (23), goats (23) and humans (13) were collected and screened for the presence of MRS. The detection of mecA gene was performed by PCR to determine the resistance of the samples to methicillin. Antimicrobial-resistance testing to penicillin, meropenem, ceftriaxone, cephalothin, oxacillin, levofloxacin, enrofloxacin, chloramphenicol, ciprofloxacin, gentamicin, clindamycin, erytromycin, linezolid, sulfamethoxazole/trimethoprim, tetracycline, doxycycline and vancomycin was performed on the mecA+ isolates. From the 161 samples, four methicillin-resistant coagulase-negative Staphylococci (MRCoNS) were isolated from human beings (31%), whereas none was isolated from animals (0%). No methicillin-resistant Staphylococcus aureus (MRSA) were isolated. All of the MRCoNS isolates from this work presented different antimicrobial resistance patterns. MRCoNS may be present in humans associated with animals while not present in the animals. Selective pressure outside of the farm and a lack of MRCoNS transmission between humans and animals may be responsible for this lack of correlation.     

Characterization of acetylcholinesterase and its molecular forms in organs of five freshwater teleosts

The activity and molecular forms of acetylcholinesterase (AChE, EC 3.1.1.7) were characterized in the brain, heart, white skeletal muscle and liver of the Siberian sturgeon (Acipenser baeri), European eel (Anguilla anguilla), wels (Silurus glanis), common carp (Cyprinus carpio) and silver carp (Hypophthalmichthys molitrix), belonging in four families. The brain had the highest (between 183.2±10.6 and 1361.2±189.7 mU mg^-1 protein and between 4.7±0.4 and 23.5±2.7 U g^-1 wet tissue) and the liver the lowest (between 25.3±1.9 and 126.5±22.7 mU mg^-1 protein and between 1.0±0.1 and 4.5±0.5 U g^-1 wet tissue) activity in all these fish, with the exception of the wels, where the heart contained the smallest amount of AChE (26.2±7.5 mU mg^-1 protein and 1.2±0.3 U g^-1 wet tissue). The highest tissue AChE activity was found in the carp and lowest in the sturgeon. The solubility properties and molecular forms of AChE in the four tissues were studied by extraction in high-salt medium (1.0 M NaCl) with and without the detergent Triton X-100 (0.5%, v/v). The proportions of detergent-soluble (DS) and salt-soluble (SS) forms of AChE varied considerably from one species to another, but a general tendency could be observed: the proportion of DS AChE was generally higher in the brain (between 70.4±4.1 and 82.5±1.2%) and lower in the heart (between 12.0±2.0 and 51.3±2.3%) and skeletal muscle (between 16.0±2.0 and 64.4±0.7%). Velocity sedimentation centrifugation revealed that most tissues contained the G₄ or A₄ form. Three tissues (brain, heart, and liver) of the Siberian sturgeon contained a relatively high proportion of the G1 form (between 24.3±2.4 and 28.6±4.9%), while the skeletal muscle of the wels contained only the most complex type of AChE, i.e., A₁₂. Amongst the tissues studied, the brain and liver mostly contained amphiphilic globular forms of the G₄ type, while the heart and skeletal muscle were rich in asymmetric forms (A₄ and A₁₂). It was concluded that the differences revealed in the activities of AChE and in the distribution of its molecular forms may be connected with the variation in behavioral habits (feeding and swimming patterns), and the developmental stage of the nervous system.     

Characterization of single‐nucleotide polymorphism markers in the Mediterranean mussel,Mytilus galloprovincialis

The Mediterranean mussel, Mytilus galloprovincialis, is one of the most important aquaculture species in Europe. Appropriate molecular markers are required to evaluate genetic resources and to trace genealogies in breeding programmes for improving mussel culture. Microsatellites have been commonly applied to this purpose in other species. However, Mediterranean mussel microsatellites have demonstrated high frequencies of null alleles that hamper accurate estimates of population parameters and confident parentage inferences. As alternative markers, we have characterized in silico 25 potential single‐nucleotide polymorphism (SNP) markers in the Mediterranean mussel from expressed sequence tag (EST) public databases. The genotyping of SNPs was performed using a single‐base extension approach. Their polymorphism was evaluated in 47 individuals from an Atlantic population. Out of the 25 potential SNPs tested, 12 were technically feasible (producing a single amplicon) and polymorphic. All were biallelic and had an unbiased heterozygosity ranging from 0.160 to 0.504. One SNP was from a mitochondrial gene. The combined potential of nuclear SNPs for parentage assignment gave an exclusion probability of a false couple of parents of 0.9471. These markers will be useful for evaluating resources and tracing genealogies in genetic breeding programmes implemented to solve the main problems of mussel culture.     

Stand structure of an uneven-aged fir–beech forest with an irregular diameter structure: modeling the development of the Belevine forest, Croatia

This study focuses on the problem of irregular diameter structure in a silver fir–beech selection (plenter) forest with a “surplus” of large diameter trees and a lack of natural regeneration and small diameter trees. We sampled 274 plots (900 m² each) in the Belevine research site (266.24 ha) in the mountain region Gorski Kotar (Croatia), where diameter (dbh) distribution, diameter increment, and natural regeneration were analyzed in detail. A low density of natural regeneration, weak annual recruitment of small (10 cm dbh) diameter trees (only five trees per hectare), delayed diameter growth of trees, and a low annual rate of trees reaching the next dbh class were attributed to the current irregular dbh structure. The stand development prediction for the next 50 years is based on a simulation model, which considers the current diameter structure, increment, recruitment, and future cutting regime. Intensive cutting in the first of five 10-year cutting cycles (intensity higher of 25%) is needed to initiate natural regeneration and to accelerate growth of young silver fir trees. In the next 50 years, the irregular diameter structure will be gradually improved.     

Back kinematics of healthy trotting horses during treadmill versus over ground locomotion

Reasons for performing study: Treadmill locomotion is frequently used for training of sport horses, for diagnostic purposes and for research. Identification of the possible biomechanical differences and similarities between the back movement during treadmill (T) and over ground (O) locomotion is essential for the correct interpretation of research results. Objectives: To compare the kinematics of the thoracolumbar vertebral column in treadmill and over ground locomotion in healthy horses. Methods: Six sound Dutch Warmblood horses trotted on a T and O during 10 s at their own preferred velocity (mean ± s.d. 3.6 ± 0.3 m/s T and 3.6 ± 0.1 m/s O), which was the same in both conditions. Kinematics of the vertebral column was captured by infrared cameras using reflective skin markers attached over the spinous processes of selected vertebrae and other locations. Flexion‐extension and lateral bending range of motion (ROM), angular motion pattern (AMP) and intravertebral pattern symmetry (IVPS) of 5 vertebral angles (T6‐T10‐T13, T10‐T13‐T17, T13‐T17‐L1, T17‐L1‐L3 and L1‐L3‐l5) were calculated. Neck angle, linear and temporal stride parameters and protraction‐retraction angles of the limbs were also calculated. Results: The vertical ROM (flexion‐extension) was similar in both conditions, but the horizontal ROM (lateral bending) of the lumbar angles T17‐L1‐L3 and L1‐L3‐L5 was less during T locomotion (mean ± s.d. difference of 1.8 ± 0.6 and 1.7 ± 0.9°, respectively, P>0.05). During O locomotion, the symmetry pattern of the lumbar vertebral angles was diminished from 0.9 to 0.7 (1 = 100% symmetry) indicating increased irregularity of the movement (P>0.05). No differences were found in the basic linear and temporal stride parameters and neck angle. Potential relevance: Vertebral kinematics during treadmill locomotion is not identical to over ground locomotion, but the differences are minor. During treadmill locomotion lumbar motion is less, and caution should be therefore taken when interpreting lumbar kinematics.     

Fluorescence of Extracts from Argentine Hake (Merluccius hubbsi) Muscle and Its Correlation with Quality Indices During Chilled Storage

Argentine hake (Merluccius hubbsi) was stored chilled for 8 days. The evolution of fluorescence of the aqueous and organic extracts of muscle during storage at 4°C was studied and correlated with common damage indices; pH, total volatile basic nitrogen, free fatty acids, and the fluorescence intensity of the aqueous phase showed a good correlation with storage time. The excitation-emission matrices of the aqueous extracts were also obtained. A fluorescence landscape that could be attributed to tryptophan decreased with storage time, meanwhile a peak (around 340 and 400 nm for excitation and emission wavelengths, respectively) is developed during chilled storage.     

Effects of dietary supplementation with probiotic live yeast Debaryomyces hansenii on the immune and antioxidant systems of leopard grouper Mycteroperca rosacea infected with Aeromonas hydrophila

Two trials were conducted to determine the efficacy of fish fed live yeast Debaryomyces hansenii strain CBS 8339 on immune and antioxidant systems in leopard grouper Mycteroperca rosacea infected with Aeromonas hydrophila. Juveniles (12±0.5 g) were fed with a control diet or a D. hansenii‐supplemented diet (10⁶ colony‐forming units per gram) for 5 weeks. The live weight of fish was registered on a weekly basis. After 4 weeks, fish from each treatment were immunocompromised with pathogenic A. hydrophila and further fed for 1 week in order to evaluate the effect on immunological and antioxidant parameters. Generally, the results showed enhanced growth performance in fish fed the diet containing yeast compared with the control. Addition of live yeast had no significant effect on the immunological parameters after 4 weeks of feeding. However, post infection with A. hydrophila fish fed the yeast‐supplemented diet resulted in a significant increase in the levels of plasmatic immunoglobulin M. Superoxide dismutase and catalase (CAT) activities were significantly higher in the yeast group. In this fish, CAT and heat shock protein 70 genes were up‐regulated before and after infection of A. hydrophila. The present study is the first one reporting that yeast (D. hansenii) can enhance immunity and resistance against A. hydrophila.     

Development of digestive biochemistry in the initial stages of three cultured Atherinopsids

The early development of the digestive biochemistry of three cultured Atherinopsids (two Odontesthes species and Chirostoma estor) was investigated. Relatively high lipolytic and high alkaline proteolytic activities were found in all species. In addition, gene expression of lipase and trypsin during the early development of C. estor not only indicates an early onset of the lipase activity but also demonstrates a major importance of protein digestion in this species. However, despite all of them share similar habitats, differences in their digestive activities were evident. The rise in the activities of brush border membrane enzymes correlated with the decrease in cytosolic activities, used as indicator of maturation of the digestive tract, was detected in both species of Odontesthes at 9 weeks after hatching. In contrast, no similar signs were measured in C. estor, this being in agreement to the long weaning period (nearly 5 months) reported for this species.     

Comparative intestinal absorption of amino acids in rainbow trout (Oncorhynchus mykiss), totoaba (Totoaba macdonaldi) and Pacific bluefin tuna (Thunnus orientalis)

The kinetics of amino acid absorption in the proximal section of three fish species were studied using an everted intestine technique and a pancreatic digest of casein (Tryptone), as the model amino acid mixture. Fresh intestine of rainbow trout (Oncorhynchus mykiss) was used to set up the experimental system, and results were compared with those obtained using intestines from totoaba (Totoaba macdonaldi) and bluefin tuna fish (Thunnus orientalis). The kinetics of amino acid absorption was sigmoidal for all amino acids and for each species of fish tested. In general, specific absorptions of essential amino acids were higher than those of non‐essential ones. No correlation between the concentration of amino acids in the Tryptone solution and the corresponding absorptions was found. The maximum specific absorption rates of all amino acids for trout were 10 times higher than those determined for totoaba and bluefin tuna. The relative amounts of the different amino acids preferentially absorbed in all three species were different. Results obtained from the everted technique may be applicable to the design of formulated diets for large fish species with commercial value.     

In silico and gene expression analysis of the acute inflammatory response of gilthead seabream (Sparus aurata) after subcutaneous administration of carrageenin

Fish Physiology and Biochemistry - Inflammation is one of the main causes of loss of homeostasis at both the systemic and molecular levels. The aim of this study was to investigate in silico the...