Chitosan Stimulates Defense Reactions in Grapevine Leaves and Inhibits Development of Botrytis Cinerea

Chitosan (β-1,4-linked glucosamine oligomer) derived from crab shells conferred a high protection of grapevine leaves against grey mould caused by Botrytis cinerea. Under controlled conditions, it was shown to be an efficient elicitor of some defense reactions in grapevine leaves and to inhibit directly the in vitro development of B. cinerea. Treatment of grapevine leaves by chitosan led to marked induction of lipoxygenase (LOX), phenylalanine ammonia-lyase (PAL) and chitinase activities, three markers of plant defense responses. Dose-response curves show that maximum defense reactions (PAL and chitinase activities) and strong reduction of B. cinerea infection were achieved with 75–150 mg l⁻¹ chitosan. However, greater concentrations of chitosan did not protect grapevine leaves with the same efficiency, but inhibited mycelial growth in vitro. Present results underlined the potency of chitosan in inducing some defense responses in grapevine leaves which in turn might improve resistance to grey mould.     

Experimental udder infection with Escherichia coli. 2. Morphological changes of the epithelium in the area of the glands of the milk cistern and the large milk ducts

Optical light, transmission, and scanning electron microscopy were used in investigations of epithelia in the glandular region of the milk cistern and greater lactiferous ducts and yielded the following findings, four and six hours from infection: degeneration and necrosis of epithelial cells, intraepithelial foreign cell infiltration (neutrophilic granulocytes, lymphocytes, macrophages), intra-epithelial oedema and locally delimited epithelial loss. The lesions differed by intensity in various regions, so that a distinction could be made among five froms of epithelial alterations. The epithelial cells revealed their capability of absorbing pathogenic material and storing it in cytoplasmic vacuoles. This was considered to reflect active involvement of epithelium in antibacterial defence, and consequently, to reflect the role played by epithelium as a defence barrier in the milk cistern and greater lactiferous ducts.     

New gene-based approaches for an AIDS vaccine

Vaccine approaches against AIDS have focused on inducing cellular immune responses, since many studies revealed the role of T cell responses in the control of human immunodeficiency virus or simian immunodeficiency virus (SIV) infections. The experimental infection of rhesus macaques with SIV or chimeric SHIV is routinely used as a model for AIDS. In such models, DNA immunization is a tool to elicit specific T cell responses and to study their protective efficacy. DNA immunogenicity in primates depends on parameters such as level of antigen expression, choice of the antigen among SIV proteins, use of fusion proteins, route of immunization, and addition of adjuvants. Recent results suggest that priming with DNA and boosting with attenuated recombinant viral vectors, each expressing corresponding SIV antigens, leads to improved specific immunity and, in some cases, affords protection against pathogenic challenge. After preclinical evaluations, DNA has entered clinical trials for a therapeutic or prophylactic gene-based AIDS vaccine.     

Chitosan oligomers and copper sulfate induce grapevine defense reactions and resistance to gray mold and downy mildew

ABSTRACT Chitosan (CHN), a deacetylated derivative of chitin, was shown to be efficient in promoting plant defense reactions. CHN oligomers of different molecular weight (MW) and degree of acetylation (DA) triggered an accumulation of phytoalexins, trans- and cis-resveratrol and their derivatives epsilon-viniferin and piceid, in grapevine leaves. Highest phytoalexin production was achieved within 48 h of incubation with CHN at 200 mug/ml with an MW of 1,500 and a DA of 20% (CHN1.5/20), while oligomers with greater MW were less efficient, indicating that a specific MW threshold could be required for phytoalexin response. Treatment of grapevine leaves by highly active CHN1.5/20 also led to marked induction of chitinase and beta-1,3-glucanase activities. CHN1.5/20 applied together with copper sulfate (CuSO(4)) strongly induced phytoalexin accumulation. CuSO(4) alone, especially at low concentrations also elicited a substantial production of phytoalexins in grapevine leaves. Evidence is also provided that CHN1.5/20 significantly reduced the infection of grapevine leaves by Botrytis cinerea and Plasmopara viticola, and in combination with CuSO(4) conferred protection against both pathogens.     

Activity of spinosad on stored-tobacco insects and persistence on cured tobacco stripst

Every year raw tobacco and manufactured tobacco products are lost to two major storage pests, the cigarette beetle, Lasioderma serricorne (F) and the tobacco moth, Ephestia elutella (Hiibner). Post-harvest management of both insects is achieved through sanitation, insect monitoring and fumigation with phosphine. However, insect resistance to phosphine and control failures have been reported, and fumigants are under constant regulatory pressure. Here we report the evaluation of spinosad, a bioinsecticide derived from the fermentation of the soil micro-organism Saccharopolyspora spinosa Mertz & Yao. Spinosad was first registered in 1997 and is now widely used as a field pest control agent on many crops, including tobacco. The insecticidal activity of the fermentation product (technical spinosad, TS) was measured by diet incorporation assays against L serricorne and E elutella larvae. Mortality levels were determined on newly hatched larvae and over the whole insect life cycle. For both species, no emergence of adult insects was observed in cured tobacco sprayed with 50mg TS kg(-1) and inoculated with eggs or newly hatched larvae. These results indicated that spinosad has potential for the control of both species in stored tobacco, since 100% control of both pests could be achieved at 50 mg TS kg(-1), and with almost full control (90-95%) at 10 mg kg(-1). We also monitored the stability of the product on cured tobacco. The original concentration of the main active component of TS, spinosyn A, did not change significantly over 18 months, indicating no loss of spinosad during a typical leaf storage period of time. Bioassays against larvae confirmed that the bioinsecticidal activity of spinosad was retained.     

Mutants in the lipopolysaccharide of Brucella ovis are attenuated and protect against B. ovis infection in mice

Brucella spp. are Gram-negative bacteria that behave as facultative intracellular parasites of a variety of mammals. This genus includes smooth (S) and rough (R) species that carry S and R lipopolysaccharides (LPS), respectively. S-LPS is a virulence factor, and mutants affected in the S-LPS O-polysaccharide (R mutants), core oligosaccharide or both show attenuation. However, B. ovis is naturally R and is virulent in sheep. We studied the role of B. ovis LPS in virulence by mutating the orthologues of wadA, wadB and wadC, three genes known to encode LPS core glycosyltransferases in S brucellae. When mapped with antibodies to outer membrane proteins (Omps) and R-LPS, wadB and wadC mutants displayed defects in LPS structure and outer membrane topology but inactivation of wadA had little or no effect. Consistent with these observations, the wadB and wadC but not the wadA mutants were attenuated in mice. When tested as vaccines, the wadB and wadC mutants protected mice against B. ovis challenge. The results demonstrate that the LPS core is a structure essential for survival in vivo not only of S brucellae but also of a naturally R Brucella pathogenic species, and they confirm our previous hypothesis that the Brucella LPS core is a target for vaccine development. Since vaccine B. melitensis Rev 1 is S and thus interferes in serological testing for S brucellae, wadB mutant represents a candidate vaccine to be evaluated against B. ovis infection of sheep suitable for areas free of B. melitensis.     

Esophageal obstruction caused by a left aortic arch and an anomalous right patent ductus arteriosus in two German Shepherd littermates

OBJECTIVE: To present details of surgical management of an unusual vascular ring anomaly in two German Shepherd littermates. STUDY DESIGN: Clinical case report. STUDY POPULATION: Three-month-old intact male and female German Shepherd littermates. RESULTS: In each dog, the esophagus was obstructed by a vascular ring comprised of the left aortic arch, an anomalous patent right ductus arteriosus, and the pulmonary artery ventrally. Surgical treatment consisted of dividing and oversewing the patent right ductus arteriosus. Neither dog has clinical signs of esophageal disease one year after treatment. CONCLUSION AND CLINICAL RELEVANCE: This vascular ring anomaly should be considered a possibility in any young dog with esophageal obstruction and a machinery murmur.     

Kinematics of whip spider pedipalps: a 3D comparative morpho-functional approach

Amblypygi are tropical and subtropical ambush predators that use elongated, raptorial pedipalps for different activities. Although pedipalp use in predation and courtship has been explored in videography in vivo analyses, kinematic ex vivo examination of these appendages has not been conducted. Here, we rectify this lack of data by using micro-CT scans to 3D-kinematically model the appendage morphology and the range of motion (ROM) of the joints for Damon medius and Heterophrynus elaphus. We illustrate the successful application of this technique to terrestrial euarthropods in determining the maximum ROM values for each pedipalp joint. We also note that, in life, these values would be lower due to motion restricting structures like tendons, arthrodial membranes, and muscles. We further compare our maximum values obtained here with data from video-based motion analyses. The ROM of each joint shows the greatest flexibility in the femur-tibia joint (140–150°), the lowest in the basitarsus-claw joint (35–40°). ROM in the tibia-basitarsus joint is markedly distinct (D. medius: 44°; H. elaphus: 105°). This disparity reflects how H. elaphus uses the joint in the capture basket, while D. medius uses the femur-tibia joint to form the capture basket. We further illustrate notable vertical motion of the H. elaphus pedipalp compared to D. medius. This difference reflects the retro-ventral trochanter apophysis of H. elaphus. Our study opens the possibility to further whip spider kinematic understanding. Examination of other taxa using this approach will result in a more comprehensive understanding of the ecological significance and ethological implications of this unique arachnid group.