Microbial origin of the abdominal swelling affecting farmed larvae of gilt-head seabream, Sparus aurata L.

The aetiological agents of the abdominal swelling affecting farmed larvae of gilt-head seabream, Sparus aurata L., were studied. Four Vibrio strains were isolated from larvae of S. aurata affected by this disease, and all strains reproduced the disease in healthy larvae under controlled infection experiments, producing a significant increase of the mortality rates compared to the control (non-inoculated larvae). Several enzymatic properties, which can act as .virulence factors, were demonstrated both in the extracellular products (ECPs) and In live cells of the strains tested. Histopathological examinations of the infected fish larvae revealed important changes of the anterior intestine and liver characterized by a marked hyperthrophy of the intestinal epithelium and hepatocytes, and by a separation of the mucosal and submucosal layers in the digestive tube. These histological alterations were associated with the constant presence of cocobacillar bacteria in the anterior intestine and in the liver. However, the precise pathogenic mechanisms of the strains tested have not been completely elucidated yet.     

Systemic accumulation of novel proteins in the apoplast of the leaves of potato plants following root invasion by the cyst-nematode Globodera rostochiensis

Systemic changes in gene expression have been analysed in potato plants following root infection with the cyst nematode Globodera rostochiensis. Root invasion by the pathogen induced major changes to occur in the composition of proteins extracted in intercellular fluid from the leaf apoplastic space. The changes involved the disappearance of innate leaf proteins and the accumulation of novel gene products. Analysis by SDS-PAGE indicated that the leaf polypeptides induced on nematode invasion are not cultivar or nematode pathotype-specific. Aspirin treatment of leaves induced a similar pattern to that induced systemically by nematodes, but several gene products were specific for the latter method of induction. In particular, two abundant [¹²⁵I]-Con A binding polypeptides of high M_r accumulated in intercellular fluid only on aspirin treatment. The results are discussed in the context of the role of the extracellular leaf proteins and their modulation by systemic signals.     

A fluoroquinolone resistance protein from Mycobacterium tuberculosis that mimics DNA

Fluoroquinolones are gaining increasing importance in the treatment of tuberculosis. The expression of MfpA, a member of the pentapeptide repeat family of proteins from Mycobacterium tuberculosis, causes resistance to ciprofloxacin and sparfloxacin. This protein binds to DNA gyrase and inhibits its activity. Its three-dimensional structure reveals a fold, which we have named the right-handed quadrilateral beta helix, that exhibits size, shape, and electrostatic similarity to B-form DNA. This represents a form of DNA mimicry and explains both its inhibitory effect on DNA gyrase and fluoroquinolone resistance resulting from the protein's expression in vivo.     

Percentage of Ubiquitinated Spermatozoa does not Correlate with Fertilizing Capacity of Thawed Bovine Semen

In the spermatozoa of some species, the ubiquitin–proteasome system detects altered proteins and tags them for elimination by the proteasome. In some species' ejaculates, a high proportion of ubiquitinated spermatozoa (i.e. those having ubiquitin bound to the altered or damaged membrane proteins) has been related to infertility. The aim of this study was to assess whether the percentage of ubiquitinated spermatozoa relates to fertility of dairy bulls and whether ubiquitination increases during protein remodelling that occurs during in vitro spermatic capacitation. Thirty‐two frozen semen straws from four high‐fertility (ReproMax^®) and four normal‐fertility (Normal) Holstein‐Friesian sires were evaluated. Ubiquitinated and capacitated spermatozoa were quantified by sperm ubiquitin tag immunoassay and chlortetracycline stain, respectively. Fertilizing capacity of sires was assessed by in vitro fertilization. No differences were found between Normal and ReproMax^® sires with regard to the observed percentage of ubiquitinated spermatozoa (42.97 ± 3.69% and 49.68 ± 9.27%, respectively; p > 0.05). Additionally, no differences were found in the percentage of ubiquitinated spermatozoa as a consequence of spermatic capacitation in either Normal (42.97 ± 3.69% before capacitation vs 44.67 ± 7.5% after; p > 0.05) or ReproMax^® sires (49.68 ± 9.27% before vs 45.05 ± 7.51% after; p > 0.05). The percentage of ubiquitinated spermatozoa in a thawed sperm samples did not correlate with its in vitro fertilizing capacity; thus, this assay does not prove useful to detect in vivo fertility differences between sires. Additionally, protein degradation occurring during remodelling of the spermatozoon plasma membrane during the capacitation process does not seem to involve the ubiquitin–proteasome system.     

Rooting potential in plum hardwood cuttings: I. Relationship with shoot diameter

The length of the annual shoots produced on hedges of the plum rootstock Prunus insititia ‘Pixy’ was determined more by the duration of their growth than by their growth rate. The most dominant shoots from the distal part of the framework grew for more than 16 weeks while weak shoots from lower down the framework stopped growing in half that time. There was a curvilinear relationship between shoot length and the proximal diameter of the cutting prepared from it, for shoots from various sources comprising different levels of winter pruning and different parts of the framework of the bush. The extent to which rooting potential was expressed was determined by the management of the rooting medium. When this was too wet, cutting bases developed necrosis which was particularly prevalent in the thinner cuttings. This problem was avoided by planting cuttings with their bases on rapidly draining sand rather than in granulated bark. Under these circumstances percentage rooting decreased with increasing cutting thickness. Before necrosis was controlled in this way rooting increased with increasing proximal stem diameter up to 6-8 mm, reflecting the susceptibility of thin cuttings to over-wet conditions.     

Antigen-specific lymphocyte proliferative responses in vaccinated and Hypoderma lineatum-infested calves.

Cattle infested with the common cattle grub, Hypoderma lineatum (Villers) develop specific humoral antibodies and a cellular immune reaction, defined by delayed-type hypersensitivity, to purified H. lineatum proteins. This investigation was designed to study the antigen-specific bovine lymphocyte response to hypodermin A (HyA), a serine protease of larval first-instar H. lineatum. Calves were vaccinated with either native or denatured HyA, and challenge-infested with H. lineatum. The kinetic development of a cellular immune response to HyA was monitored during vaccination and infestation. The HyA-specific responses were highly variable and weak during vaccination and infestation. Although HyA-specific lymphocyte blastogenic responses were observed, no correlation was noted between the magnitude of antigen-specific, peripheral lymphocyte proliferation and larval mortality. In striking contrast to responses observed during infestation, intense HyA-specific lymphocyte responses were observed with 3 calves 6 months after recovery from infestation. In addition, those responses were further heightened by a 250 micrograms booster injection of pure HyA.     

Polymorphism of the CD4 and CD5 differentiation antigens in cattle.

Monoclonal antibodies (mAbs) specific for bovine CD4 and CD5 antigens have been found to identify polymorphic determinants on these molecules. In the case of CD5, mAb IL-A67 recognises one allotypic form of the antigen while four other CD5-specific mAbs in the workshop (CC17, CC29, BLT-1 and 8C11) recognise a second allotype. The CD4-specific mAbs submitted to the workshop reacted with the cells of all animals tested. However, a further two mAbs (CC26 and IL-A18) specific for CD4 were found to react with cells only from about 85% of animals tested. Sequential immuno-precipitation experiments together with family studies showed that the allotypes of CD4 and CD5 are both inherited in a simple Mendelian manner and are co-dominantly expressed. One of the CD5 allotypes was not detected in Bos taurus animals while the gene frequency of the second allotype was only about 10% in the B. indicus animals tested. The gene frequency of the CD4 allotype detected by CC26 and IL-A18 was similar in the two sub-species.     

Assessment of Suitable Porcine Semen for Freezing,According to the Ejaculate Characteristics in the Iberico × Landrace Breed

A total of 35 ejaculates were studied in order to assess the suitability of porcine semen for freezing according to the ejaculate characteristics. The effects of the freezing procedure were identified; a decrease in motility and acrosome quality was found after thawing. The best results on motility were linked to the ejaculates with a volume of less than 100 ml of the sperm‐rich fraction, a concentration lower than 450 × 10⁶ spermatozoa/ml and an agglutination score below 2. However, the best normal apical ridge (NAR) was found when the volume of the sperm‐rich fraction was greater than 150 ml. For this reason, an intermediate volume of the sperm‐rich fraction of the ejaculate for the best motility and the best NAR, a concentration lower than 450 × 10⁶ spermatozoa/ml and a rate of agglutination below 2 should provide the best quality after freezing. This study also attempted to determine whether a positive effect of ejaculate selection on the overall freezing performance might be expected.     

Further sources of resistance toHeterodera rostochiensis woll. in the andigena potatoes

Summary The Andigena potatoes (Solanum tuberosum ssp.andigena) of the Commonwealth Potato Collection (C.P.C.) were screened for resistance to non-pathotype A populations ofHeterodera rostochiensis. Three populations were used: Duddingston (pathotype B), Nocton (pathotype C) and Changed Little Ouse (pathotype C). Resistance was found in C.P.C. 2775, 2802 and 2805. C.P.C. 2775 also had resistance to pathotype A (Feltwell population) and appeared to have two dominant genes,H ₁ giving resistance to pathotype A andH ₃ giving resistance to pathotypes B and C. C.P.C. 2802 was susceptible to pathotype A and appeared to have only anH ₃-type gene. C.P.C. 2805, which has not been investigated thoroughly because the plants lacked vigour, may also have anH ₁ and anH ₃ gene. The value of C.P.C. 2775, 2802 and 2805 for breeding nematode-resistant potatoes will only be known when they have been tested with further non-pathotype A populations and when the genetics of resistance have been investigated thoroughly.