Pseudorabies virus infection in mink: a host-specific pathogenesis

Pseudorabies virus (PRV) is an alphaherpesvirus that causes a neurological disease in many wild and domestic animals. The neuropathology elicited by PRV is quite consistent regardless of the host with the only exception of mink, in which it is characterized by a vasculopathy rather than by an encephalitis. In this study, we aimed to investigate the underlying pathogenic mechanism(s) of PRV infection in mink by using immunohistochemistry and laser capture microdissection (LCM) on material from naturally and experimentally infected animals. The inflammatory reaction induced by PRV was minimal or absent not only in the nervous system, where we identified a low number of macrophages and a few T lymphocytes, but also in the primary replication site, the oropharyngeal mucosa; however, the number of PRV-infected cells detected by immunohistochemistry was extremely high both in the peripheral mucosa and in the nervous tissue. On the other hand, the vascular pathology included parenchymal hemorrhages of various degrees and, in specific cortical areas of the brain, fibrinoid degeneration of the capillary walls. Detection of viral antigens by immunohistochemistry revealed infection of endothelial cells of capillaries situated both in the oropharyngeal mucosa and in the brain stem; the presence of PRV DNA in vessels was further demonstrated by PCR performed on LCM samples of brain capillaries. These results can be interpreted as supporting the idea that the different pathology of the disease in mink may be the consequence of an increased endotheliotropism of PRV in this species. Infection of the vessel wall may then lead to vascular pathology and impairment in endothelial cell function, resulting in a weak immune response to infection.     

Validation of an ELISA method for the serological diagnosis of canine brucellosis due to Brucella canis

In the present study, the validation of an enzyme-linked immunosorbent assay (ELISA) for serodiagnosis of canine brucellosis is described. Two different antigenic extracts, obtained by heat or ultrasonic homogenization of microbial antigens from a wild isolate of Brucella canis bacteria, were compared by ELISA and Western blot (WB). A total of 145 canine sera were used to define sensitivity, specificity and accuracy of the ELISA as follows: (1) sera from 34 animals with natural B. canis infection, confirmed by blood culture and PCR, as well as 51 sera samples from healthy dogs with negative results by the agar–gel immunodiffusion (AGID) test for canine brucellosis, were used as the control panel for B. canis infection; and (2) to scrutinize the possibility of cross reactions with other common dog infections in the same geographical area in Brazil, 60 sera samples from dogs harboring known infections by Leptospira sp., Ehrlichia canis, canine distemper virus (CDV), Neospora caninum, Babesia canis and Leishmania chagasi (10 in each group) were included in the study. The ELISA using heat soluble bacterial extract (HE-antigen) as antigen showed the best values of sensitivity (91.18%), specificity (100%) and accuracy (96.47%). In the WB analyses, the HE-antigen showed no cross-reactivity with sera from dogs with different infections, while the B. canis sonicate had various protein bands identified by those sera. The performance of the ELISA standardized with the heat soluble B. canis antigen indicates that this assay can be used as a reliable and practical method to confirm infection by this microorganism, as well as a tool for seroepidemiological studies.     

Monitoring neonicotinoid resistance in biotype B of Bemisia tabaci in Florida

BACKGROUND: Biotype B of the sweetpotato whitefly, Bemisia tabaci (Genn.), is a worldwide pest that has developed resistance to many insecticides, including the neonicotinoid class. Florida field populations were monitored for susceptibility to the neonicotinoids imidacloprid and thiamethoxam using a cut leaf petiole bioassay method. RESULTS: Average RR₅₀ values for imidacloprid increased from 3.7 in 2000 to 12.0 in 2003; decreased to 5.0 and 2.5 in 2004 and 2005, respectively; and then increased to 26.3 and 23.9 in 2006 and 2007, respectively. Populations with RR₅₀ values of about 50 to 60 during generation one reverted to RR₅₀ values of ?4 in six generations, when reared without further exposure to imidacloprid. Average RR₅₀ values for thiamethoxam increased from 2.0 in 2003 to 24.7 in 2006 and decreased to 10.4 in 2007. Populations with RR₅₀ values of about 22, 32 and 53 during generation one declined to 8, 5 and 6, respectively, after being reared for five generations without exposure to thiamethoxam. The correlation coefficient from the 26 populations that were bioassayed both with imidacloprid and thiamethoxam showed a significant positive correlation (R² = 0.58) between these populations. CONCLUSION: The high level of RR₅₀ values to imidacloprid and thiamethoxam suggest an unstable decline in the susceptibility of B. tabaci to imidacloprid and thiamethoxam, with possible cross‐resistance or predisposition for dual resistance selection. Copyright © 2009 Society of Chemical Industry     

Induction of resistance to Sclerotium rolfsii in different varieties of onion by inoculation with Trichoderma asperellum

We evaluated the ability of Trichoderma asperellum Samuels, Lieckfeldt & Nirenberg to induce resistance to the fungal plant pathogen, Sclerotium rolfsii Saccardo, in three onion (Allium cepa L.) varieties. Both the severity of disease and the activities of glucanase, chitinase and peroxidase (enzymes involved in plant resistance) were evaluated in onions inoculated with T. asperellum alone, S. rolfsii alone, or both T. asperellum and S. rolfsii (dual-inoculation) and compared to uninoculated (control) plants. In dual inoculations, the presence of T. asperellum reduced the severity of disease symptoms caused by S. rolfsii. Inoculation with T. asperellum alone increased glucanase, chitinase and peroxidase activity in bulbs, roots and leaves of all three onion varieties compared to uninoculated controls; bulbs of the variety Red Satan (RS) had the highest enzyme activity. In plants inoculated with S. rolfsii alone, enzyme activity was only increased in bulbs and roots compared to uninoculated controls. The highest levels of enzyme activity also occurred only in bulbs and roots of plants that had been dual-inoculated with T. asperellum and S. rolfsii. Plants of the RS variety showed the highest enzyme activities (both constitutive and induced) and showed the lowest severity of disease. Therefore, application of T. asperellum has potential as a biological control alternative to synthetic fungicides for protection of onion crops against infection by S. rolfsii. This protection depends on both constitutive and induced defence responses and varies amongst onion varieties.     

Biological control ofBotrytis cinerea in residues and flowers of Rose (Rosa hybrida)

Microbial isolates from living petals, petal residues and leaf residues of rose, and from laboratory collections, were evaluated for control ofBotrytis cinerea in rose. In leaf residues artificially infested withB. cinerea, isolates of the filamentous fungiGliocladium roseum, FR136 (unidentified) andTrichoderma inhamatum reduced sporulation of the pathogen by >90%, other filamentous fungi were 25–90% effective, and those of yeasts and bacteria were <50% effective. In artificially inoculated petal residues, no microbe reduced sporulation ofB. cinerea by >75%, but isolates ofCladosporium oxysporum and four yeasts were 51–75% effective, and three filamentous fungi, eight yeasts andBacillus subtilis isolates were 26–50% effective. Isolates ofT. inhamatum, C. oxysporum andG. roseum performed best againstB. cinerea among isolates evaluated in leaf residues naturally infested with the pathogen and indigenous microorganisms. Totals of ten isolates of filamentous fungi (includingC. oxysporum andC. cladosporioides), two of yeasts and five ofBacillus subtilis completely prevented lesion production byB. cinerea in detached petals, and a further six isolates of filamentous fungi (includingG. roseum) and six yeasts were 90–99% effective. Isolates ofC. oxysporum, C. cladosporioides andB. subtilis, the most effective microorganisms againstB. cinerea in flower buds, reduced number of lesions in the range of 42–65% compared with 59–89% for à standard fungicide (vinclozolin). It is suggested that application of leading antagonists Jo living rose leaves and flowers should optimize control of inoculum production byB. cinerea when the tissues die. Optimal biocontrol of lesion production in flower buds requires a better understanding of the microenvironment of petals.     

Zinc phosphate protects tomato plants against Pseudomonas syringae pv. tomato

Journal of Plant Diseases and Protection - The purpose of this study was to determine whether zinc phosphate treatments of tomato plants (Solanum lycopersicum L.) can attenuate bacterial speck...     

Evaluation,tissue culture propagation,and dissemination of ‘Saba’ and ‘Pelipita’ plantains in Costa Rica

Fruit characteristics of the disease-resistant bluggoe-type (ABB) cooking bananas (Musa × paradisiaca L.) ‘Saba’ and ‘Pelipita’ were compared with those of the susceptible ‘Currare’ (‘Horn’). Yields of ‘Saba’ and ‘Pelipita’ were similar to those of ‘Currare’; however, ‘Saba’ and ‘Pelipita’ yielded fewer hands with a greater number of small fruit when compared with ‘Currare’. Shoot-tip cultures of both clones were readily initiated on a modified Murashige and Skoog (MS) basal medium supplemented with N⁶-benzyladenine (BA) in combination with indole-3-acetic acid (IAA). Propagation cultures were initiated by splitting shoot tips along their longitudinal axis and re-culturing the individual pieces to basal medium supplemented with 5 mg l^?1 BA. Transfer of axillary shoots to hormone-free medium resulted in rapid and extensive root formation. Plantlet survival after transfer to methyl bromide-treated soil exceeded 90%. Establishment in the field was achieved following procedures normally used for vegetative propagation of this crop. Trial plantings of in vitro propagated ‘Saba’ and ‘Pelipita’ were established in various provinces of Costa Rica for grower evaluation and for future comparison of growth and reproductive development with plants of these and other cultivars propagated from corms.     

Comparison of levels and duration of detection of antibodies to bovine viral diarrhea virus 1, bovine viral diarrhea virus 2, bovine respiratory syncytial virus,bovine herpesvirus 1, and bovine parainfluenza virus 3 in calves fed maternal colostrum or a colostrum-replacement product

Colostrum-replacement products are an alternative to provide passive immunity to neonatal calves; however, their ability to provide adequate levels of antibodies recognizing respiratory viruses has not been described. The objective of this study was to compare the serum levels of IgG at 2 d of age and the duration of detection of antibodies to bovine viral diarrhea virus 1 (BVDV-1), bovine viral diarrhea virus 2 (BVDV-2), bovine respiratory syncytial virus (BRSV), bovine herpesvirus 1 (BHV-1), and bovine parainfluenza virus 3 (BPIV-3) in calves fed maternal colostrum (MC) or a colostrum replacement (CR) at birth. Forty newborn male Holstein calves were assigned to the CR or the MC group. Group CR (n = 20) received 2 packets of colostrum replacement (100 g of IgG per 470-g packet), while group MC (n = 20) received 3.8 L of maternal colostrum. Blood samples for detection of IgG and virus antibodies were collected from each calf at birth, at 2 and 7 d, and monthly until the calves became seronegative. Calves in the MC group had greater IgG concentrations at 2 d of age. The apparent efficiency of absorption of IgG was greater in the MC group than in the CR group, although the difference was not significant. Calves in the CR group had greater concentrations of BVDV neutralizing antibodies during the first 4 mo of life. The levels of antibodies to BRSV, BHV-1, and BPIV-3 were similar in the 2 groups. The mean time to seronegativity was similar for each virus in the 2 groups; however, greater variation was observed in the antibody levels and in the duration of detection of immunity in the MC group than in the CR group. Thus, the CR product provided calves with more uniform levels and duration of antibodies to common bovine respiratory viruses.     

A practical and reliable procedure for in vitro induction of tetraploid tomato

A relatively rapid, practical and reliable procedure for in vitro production of tetraploid tomato using colchicine is described. Using this procedure 11.11% generation of tetraploids was obtained after exposure of seedling shoot meristem explants with 8 mM colchicine for 96 h. Confirmation of tetraploid production (2n = 48), was determined by flow cytometry and verified by cytogenetic analysis of root tip preparations. The results indicate that the procedure is adequate for induction and screening of tetraploid tomato plantlets.