Bacterial host interaction of GFP‐labelled Vibrio anguillarum HI‐610 with gnotobiotic sea bass,Dicentrarchus labrax (L.), larvae

The location and cell damage caused by Vibrio anguillarum, the causative agent of classical vibriosis, within the developing gut of the newly hatched sea bass, Dicentrarchus labrax (L.), is unknown. A gnotobiotic sea bass model was used to investigate the early interactions of V. anguillarum with sea bass larvae. In the present study, germ‐free sea bass larvae were orally exposed to a V. anguillarum HI‐610 pathogen labelled with the green fluorescent protein (GFP‐HI‐610) and sampled at regular intervals. Pathogenic colonization of gut enterocytes was observed 2 h post‐exposure (p.e.) and onwards, whereas bacteria within the swim bladder were visualized 48 h p.e and onwards. Ultrastructural findings demonstrated direct bacterial contact with the host cell in the oesophageal mucosa and putative attachment to microvilli of mid‐ and hindgut enterocytes. The present findings form a starting point for studies assessing the impact of potential candidates (probiotics, prebiotics, antimicrobial peptides) to mitigate bacterial virulence.     

<Emphasis Type="Italic">Alternaria alternata</Emphasis> apple pathotype (<Emphasis Type="Italic">A. mali</Emphasis>) causes black spot of European pear

A disease caused by Alternaria alternata occurred on the leaves of European pear cultivar Le Lectier in Niigata Prefecture, Japan, and was named black spot of European pear. In conidial inoculation tests, the causal pathogen induced not only small black lesions on the leaves of European pear cultivar Le Lectier, but severe lesions on the leaves of apple cultivar Red Gold, which is susceptible to the A. alternata apple pathotype (previously called A. mali) causing Alternaria blotch of apple. Interestingly, the apple pathotype isolate showed the same pathogenicity as the European pear pathogen. HPLC analysis of the culture filtrates revealed that A. alternata causing black spot of European pear produced AM-toxin I, known as a host-specific toxin of the A. alternata apple pathotype. AM-toxin I induced veinal necrosis on leaves of Le Lectier and General Leclerc cultivars, both susceptible to the European pear pathogen, at 5?×?10^?7 M and 10^?6 M respectively, but did not affect leaves of resistant cultivars at 10^?4 M. PCR analysis with primers that specifically amplify the AM-toxin synthetase gene detected the product of expected size in the pathogen. These results indicate that A. alternata causing black spot of European pear is identical to that causing Alternaria blotch of apple. This is the first report of European pear disease caused by the A. alternata apple pathotype. This study provides a multiplex PCR protocol, which could serve as a useful tool, for the epidemiological survey of these two diseases in European pear and apple orchards.     

Fatty acid profile of hair lambs and their crossbreds slaughtered at different weights

This study aims to investigate the effect of breed and slaughter weight on fatty the acid profile of the Longissimus dorsi muscle in lambs. Twenty-four Santa Inês (SI), 24.5 Ile de France × 0.5 Santa Inês (Ile × SI) and 12.5 Texel × 0.5 Santa Inês (Te × SI), slaughtered at different weights (30, 35, 40, and 45 kg), were evaluated. The animals were reared with creep feeding to weaning and were feedlot finished, receiving a diet composed of 30% hay and 70% concentrate. The analyses of fatty acids were carried out on the Longissimus muscle of the 13th rib. The experiment was in a three by four factorial design. The total mean saturated fatty acid level was 44.88%, with 43.30% monounsaturated fats and 1.72% polyunsaturated. The major fatty acids found included oleic (43%), palmitic (22%), and stearic (18%). The concentration of desirable fatty acids varied from 61.56% to 66.78%, with Te × SI (66.78%) having the highest levels (P < 0.05). The slaughter weight affected (P < 0.05) the saturated and unsaturated (both mono and poly) fatty acid profiles. The 35-kg slaughter weight showed the most desirable fatty acid profile. The saturated myristic and stearic fatty acids decreased with an increase in the slaughter weight, but oleic acid increased. In the conditions of the present study, the meat from Santa Ines and Texel × Santa Ines lambs had fatty acid profiles more beneficial for human health due to the higher levels of unsaturated fatty acids.     

Stem cell factor supports migration in canine mesenchymal stem cells

Adult Mesenchymal Stem Cells (MSC) are cells that can be defined as multipotent cells able to differentiate into diverse lineages, under appropriate conditions. These cells have been widely used in regenerative medicine, both in preclinical and clinical settings. Initially discovered in bone marrow, MSC can now be isolated from a wide spectrum of adult and foetal tissues. Studies to evaluate the therapeutic potential of these cells are based on their ability to arrive to damaged tissues. In this paper we have done a comparative study analyzing proliferation, surface markers and OCT4, SOX9, RUNX2, PPARG genes expression in MSC cells from Bone marrow (BMMSC) and Adipose tissue (ASC). We also analyzed the role of Stem Cell Factor (SCF) on MSC proliferation and on ASCs metalloproteinases MMP-2, MMP-9 secretion. Healthy dogs were used as BMMSC donors, and ASC were collected from omentum during elective ovariohysterectomy surgery. Both cell types were cultured in IMDM medium with or without SCF, 10% Dog Serum (DS), and incubated at 38 °C with 5% CO2. Growth of BMMSCs and ASCs was exponential until 25–30 days. Flow citometry of MSCs revealed positive results for CD90 and negative for CD34, CD45 and MCH-II. Genes were evaluated by RT-PCR and metalloproteinases by zymografy. Our findings indicate morphological and immunological similarities as well as expression of genes from both origins on analyzed cells. Furthermore, SCF did not affect proliferation of MSCs, however it up-regulated MMP-2 and MMP-9 secretion in ASCs. These results suggest that metalloproteinases are possibly essential molecules pivoting migration.     

Mild Drought Stress‐Induced Changes in Yield,Physiological Processes and Chemical Composition in Festuca,Lolium and Festulolium

The impact of mild drought stress (3 weeks at 40 % field water capacity) on yield, physiological processes, accumulation of proline and phenolic compounds and forage quality parameters in forage grasses was evaluated in pot experiments. During four different growing periods, the effects of water deficit were assessed in nine varieties from five species (Lolium perenne, Lolium multiflorum, Festuca pratensis, Festuca arundinacea and Festulolium braunii). All measured parameters were affected by drought stress in the different cuts. Photosynthesis, transpiration rate, stomatal conductance and dry matter yield were significantly lower in drought stress than under well‐watered conditions in all varieties. Higher water‐use efficiency was only observed during the first and fourth drought period, while this was not the case in the second and third. Mild drought stress significantly increased the content of proline, phenolic acid, flavonoids, water‐soluble carbohydrates and protein. All tested grasses showed also an increase of organic matter digestibility and cell wall digestibility under drought stress conditions.     

Deadwood volume in strictly protected,natural, and primeval forests in Poland

Standing and downed deadwood at different stages of decay provides a crucial habitat for a wide range of organisms. It is particularly abundant in unmanaged forests, such as strictly protected areas of national parks and nature reserves. The present work used the available data for such sites in Poland, analyzing a total of 113 studies concerning 79 sites to determine the causes contributing to variation in deadwood volume based on the duration of conservation, changes in deadwood volume over time (for those sites which were examined multiple times), elevation above sea level, forest type, stage of forest development, input of dead trees from the years preceding deadwood measurements, live tree volume, and the proportion of downed to standing deadwood). Depending on species composition and site altitude, most tree stands fell into one of four categories: subalpine spruce forests, montane beech-fir forests, low altitude beech-fir forests, or oak-hornbeam and riparian forests. The mean deadwood volume for all forest types amounted to 172.0 m³/ha. The mean volume of deadwood in montane beech-fir forests (223.9 m³/ha) was statistically significantly greater than in the other three forest types, for which it ranged from 103.5 to 142.5 m³/ha. A direct effect of the duration of conservation on deadwood volume was not identified. Nevertheless, analysis of repeated measurements on the same sample plots at 10-year intervals showed a consistent rise in mean deadwood volume. A linear regression model for all the analyzed factors reported from montane beech-fir forests and subalpine spruce forests showed that in addition to site altitude, another statistically significant variable was the input of dead trees (R²?=?63.54%).     

Virulence factors in Escherichia coli isolated from the blood of bacteremic neonatal calves

Twenty-five Escherichia coli isolates from the blood of critically ill bacteremic calves sampled in two separate studies on a calf-rearing farm housing over 15,000 calves, in the San Joaquin Valley, California were studied.Isolates were characterized for O serogroups and for pathotypes as determined by the presence of specific virulence factors including heat-labile enterotoxin (LT), heat-stable enterotoxins a and b (STa, STb), verotoxins 1 and 2 (VT1, VT2), cytotoxic necrotizing factor (CNF), aerobactin, intimin Eae and P, F17 and CS31A fimbrial adhesins, and resistance to bactericidal effects of serum.These isolates constituted a heterogeneous group. However, isolates were mostly aerobactin positive and often resistant to the bactericidal effects of serum. Isolates of pathotypes O78 (n=6), O119:CS31a (n=3), and P positive but O non-typeable (n=3) were associated with a high mortality rate. The remaining isolates belonged to diverse pathotypes, often possessing the adhesins P, F17, CS31A and Eae but belonging to O serogroups other than O78 and O119, and were less frequently associated with mortality.Although no virulence factor common to all isolates was identified, the capacity to use iron by the presence of aerobactin which is important to the capture of iron was a predominant factor. Moreover, certain pathotypes appear to be associated with primary colisepticemia whereas other pathotypes may cause a bacteremia without necessarily leading to septicemia.     

PEG bulking of wood cell walls as affected by moisture content and nature of solvent

Movement of polyethylene glycol (PEG) 1000 from wood cell lumens into adjacent cell walls was studied for different cell wall moisture contents (MCs) and PEG solvents; relative amounts of PEG that penetrated into the cell walls were estimated by Raman microscopy. It was shown that samples treated with PEG from chloroform solution adsorb less PEG than samples treated with PEG-water solution even in the case of samples with open cell wall structure near fibre saturation point MC. Reconditioning of PEG-chloroform treated samples at high relative humidities enhanced PEG bulking of cell walls, but not to the maximum amounts observed with PEG-water treatments. Raman microscopy was a useful tool for estimating relative PEG amounts in the cell walls.     

Two spawning stocks of icefish <Emphasis Type="Italic">Neosalanx taihuensis</Emphasis> revealed from annual reproductive cycle analyses

Neosalanx taihuensis were sampled from the Tian-e-zhou Oxbow from March 2006 through November 2007. Two separate spawning seasons were identified based on the annual reproductive cycles of the females, designated as the autumn-spawning season and the spring-spawning season. Lifespan of the offspring of the spring-spawning fish was 1 year, with them dying after the subsequent spring spawning. Autumn-spawned females seem to be the offspring of the spring-spawning fish, based on monthly changes in the length–frequency distributions. Spring-mature females had higher absolute fecundity, gonadosomatic index, and relative condition factor in 2007 than in 2006. Relative condition factor of the immature female offspring of spring-spawning fish was higher in 2007 than in 2006, portending a further increase in reproductive investment during the spring spawning of 2008. The increasing reproductive investment suggests that the population of N. taihuensis in the Tian-e-zhou Oxbow may be recovering from its recent decline.