Influence of variety and year of wheat cultivation on the chemical composition of starch and properties of glucose hydrolysates

The effect of chemical composition of starch samples derived from selected wheat cultivars harvested in different years (lipid content, amylose–lipid complex (AML) content, amylose concentration and the degree of starch damage) on physicochemical properties of glucose syrups produced from these starches was determined. It was found that glucose syrups obtained from the same wheat cultivar but harvested in different years displayed different physicochemical attributes such as concentration of reducing compounds, rate of filtration and coefficients of colour and transparency. Besides, the analyses showed that AML changed their form from amorphous to crystalline during enzymatic starch liquefaction by α-amylase. This in turn rendered starch less susceptible to enzymatic digestion and affected physicochemical features of glucose syrups. Presented results provide evidence that not only the total concentration of lipids and AML but also the AML polymorph present in starch hydrolysates (on completion of starch liquefaction) have an impact on quality of glucose syrups.     

Molecular cloning and expression analysis of the main gliadin-degrading cysteine endopeptidase EP8 from triticale

During the development and maturation of cereal grains, storage proteins are accumulated in the starchy endosperm. The enzymes responsible for the mobilisation of stored proteins during seed germination are cysteine endopeptidases and serine carboxypeptidases. In our previous study, we purified the major gliadin-degrading cysteine, endopeptidase EP8, from germinating triticale kernels. We confirmed in an experiment with exogenous gibberellic acid (GA₃) that this enzyme is synthesised in aleurone during germination. In this study, the nucleotide sequence of EP8, a barley EP-A (HvEPA) homologue, and the expression pattern during grain development and germination have been analysed. Additionally, by comparing the activity of purified EP8 with the activity pattern of crude enzymatic extracts, the lack of enzyme activity in some tissues and during certain developmental stages has been demonstrated. The correlation of the results of the EP8 expression analysis with the activity pattern of this endopeptidase allowed us to formulate a hypothesis regarding the mechanism of EP8 activity regulation. We believe that the absence of active EP8 until the third day of germination may be associated with high levels of endogenous cystatin TrcC-4, which has the ability to inhibit EP8 in vitro.     

Inhibition of Armillaria and Heterobasidion growth by Penicillium adametzii isolated from Pinus sylvestris forest soil

Penicillium adametzii significantly inhibited the growth of Heterobasidium annosum sensu stricto on 2% potato dextrose agar after 10 days at 20–25°C. Chloroform extracts from P. adametzii culture filtrate and from culture grown on rice often decreased the dry weight of Armillaria colony and diameter of Heterobasidion colony in vitro. The effect depended on species and isolate of Armillaria, Heterobasidion and P. adametzii and concentration of the extract. Higher concentration of chloroform extract increased the dry weight of stems and roots of Pinus sylvestris plants inoculated with either of two isolates of Armillaria ostoyae and one isolate of Armillaria gallica. Lower concentrations of chloroform extract increased the dry weight of stems and roots of P. sylvestris inoculated with another isolate of A. gallica. Culture filtrate of P. adametzii decreased the length of necrosis on P. sylvestris stem inoculated with either of two of four isolates of H. annosum s.s., but if applied 1 month before inoculation, it increased the length of necrosis caused by another H. annosum s.s. isolate. Twenty fractions from the P. adametzii chloroform extract were separated by thin layer chromatography. Carboxylic acids, flavonoids and glycosides were detected in chloroform extract of P. adametzii by high performance liquid chromatography/mass spectrometer analysis. This is the first report of interactions between P. adametzii and either Armillaria or Heterobasidion with P. sylvestris.     

Epidemiological features of Morel's disease in goats

Morel's disease caused by Staphylococcus aureus subsp. anaerobius was diagnosed for the first time in Poland in October 2006 in a goat flock. A second infected flock was found two months later. The course of the disease in both flocks was observed for 15-17 months. Clinical manifestation was confined to abscesses located near major superficial lymph nodes, mostly: superficial cervical, subiliac, parotid and mandibular. At necropsy no other lesions were found. The incubation period was estimated at 3 weeks. Clinical signs were seen both in young and adult goats and up to 7 abscesses in one animal were noted. Abscesses tended to persist for 1 to 5 months, then rupture and heal completely. The initial high in-flock point prevalence in both flocks (93.6% and 84.4%) dropped to approximately 10-30% during next 3-4 months. Until the end of the observation period the in-flock point prevalence remained at this level and only single abscesses were observed, mainly in young animals. No influence of the concurrent caprine arthritis encephalitis virus (CAEV) infection on the clinical course of Morel's disease was noticed. It is to be concluded that the clinical course of Morel's disease in a goat flock resembles caseous lymphadenitis (CLA). However, in Morel's disease abscesses occur more frequently in young goats and are located near, not inside, the lymph nodes, as in the case with CLA. Also, the incubation period of Morel's disease seems to be shorter (3 weeks versus 2-6 months in CLA).     

Evaluation of the influence of meloxicam and flunixin meglumine on the apoptosis of peripheral blood CD4+ and CD8+ T cells in calves

The aim of the study was to determine whether treatment with recommended doses of meloxicam or flunixin had an effect on the apoptosis of peripheral blood T lymphocytes in calves. The study was carried out on 4-5 months old calves (n = 24, 8 per group). Experimental animals were injected subcutaneously with a single dose of 0.5 mg x kg(-1) of meloxicam or intravenously with 3 doses of 2.2 mg x kg(-1) day(-1) of flunixin. The non-treatment animals served as control. Blood samples were taken at day 0 and at days 1, 2, 3, 5, 7 and 14 after the first NSAIDs injection. Apoptosis was determined by flow cytometry using Annexin V-PE/7-AAD staining. The kinetic analysis of apoptosis in the total lymphocyte population, as well as in the CD4+ and CD8+ subsets did not reveal significant differences in the frequency of early apoptotic cells between control and experimental groups throughout the period studied. Although, 24 h after administration of the first dose of NSAIDs, late-stage apoptosis/necrosis was significantly increased in the total lymphocyte population (the meloxicam group), as well as in the CD4+ (the meloxicam group and the flunixin group) and CD8+ (the flunixin group) subsets of T cells. However, this disturbance was transient, relatively poorly expressed and, thus, unlikely to be of clinical significance. Our results indicate that the use of meloxicam or flunixin in accordance with the recommended dosage regimen in cattle do not have a clinically significant influence on apoptosis of peripheral blood T cells.     

Abundance of some skeletal muscle mitochondrial proteins is associated with increased blood serum insulin in bovine fetuses

The aim of this study was to investigate the evolution of the abundance of cytochrome oxidase c subunit IV (NCOIV) and beta subunit of ATP synthase (β-ATP) during the last third of gestation in bovine skeletal muscles. Semitendinosus, longissimus thoracis and rectus abdominis muscles were chosen for the immunoblotting of the respective protein levels. Muscle and blood samples from bovine fetuses of randomly selected breeds were collected at 180, 210, and 260 days post-conception (dpc). The muscle tissue expressions of NCOIV, β-ATP were compared to blood glucose and insulin. At 260 dpc, protein levels of NCOIV raised in skeletal muscles. Additionally, β-ATP in semitendinosus and longissimus thoracis were elevated and paralleled by higher concentrations of blood serum insulin. It corroborates our previous observations indicating that accelerated metabolic differentiation of bovine skeletal muscles is associated with elevated blood insulin and occurs during the last trimester of gestation. Our observations point to the connection between insulin-sensitivity and the molecular mechanisms of mitochondrial contribution to ontogenesis of skeletal muscles.     

Horse infection with intestinal helminths in relation to age, sex, access to grass and farm system

A 11-year survey on helminth intestinal infection of horses was done in seven provinces of southern Poland. 21,641 faecal samples were collected from horses of different age, sex, breed and management system originating from 84 farms. Age was the major factor influencing the cyathostome egg excretion, the maximum being observed in yearlings and second year horses. Parascaris was found mostly in foals and yearlings. The main factor for Anoplocephala was the access to pasture. The level of cyathostome egg shedding (EPG) could be related to sex (geldings had higher values of EPG), breed (Thoroughbred had higher EPG than other breeds), and type of farm (large farms had higher EPG than others). The change during the survey of anthelmintic treatment (from benzimidazole or pyrantel embonate to avermectins) was associated with lower cyathostome EPG, and this was probably due to partial resistance to benzimidazole. The seasonality of egg shedding (lowest from November to January included) was due to management (leaving the pasture to stable, anthelmintic treatment in October). The influence of climate on EPG was significant but small, annual rainfalls and average temperature being associated negatively and positively, respectively, with high cyathostome EPG.     

Sperm quality and selected biochemical markers of seminal plasma at the beginning of the reproductive period of common carp, Cyprinus carpio L.

Changes in semen quality and selected biochemical markers were analyzed during a week of spawning season of common carp Cyprinus carpio L. Semen was obtained twice, on May 30 and on June 7, and each time it was collected 24 h after hormonal stimulation using Ovopel [(d-Ala⁶, Pro⁹ NEt)-mGnRH + metoclopramide] in 1 pellet kg^?1. The total volume of semen (ml), volume of semen per kg of body weight (ml kg^?1 b.w.), sperm concentration (×10⁹ ml^?1), total number of sperm per kg of body weight (×10⁹ kg^?1 b.w.), pH of semen, pH of seminal plasma, seminal plasma osmotic pressure (mOsm kg^?1) and the total protein content in seminal plasma (mg ml^?1) were determined. A 10 mM Tris buffer containing 100 mM NaCl with 0.5 % BSA (pH 9.0, osmolality 200 mOsm kg^?1) was used to activate sperm. The following computer-assisted sperm analysis (CASA) parameters were determined: percentage of motile sperm (MOT, %), progressively motile sperm (PRG, %), curvilinear velocity (VCL, μm s^?1), straight-line velocity (VSL, μm s^?1), movement linearity (LIN, %), wobbling index (WOB, %), amplitude of lateral head displacement (ALH, μm) and beat cross-frequency (BCF, Hz). The volume of semen per kg of BW, total number of sperm per kg of BW and semen pH were significantly lower at the second semen sampling compared to the first semen sampling. Volume of semen at the second sampling correlated positively with CASA parameters. A lack of differences among CASA parameters between both collection periods indicates good quality of carp sperm hormonally stimulated with Ovopel twice at a 1-week interval.