Effect of an immunomodulator on the efficacy of an attenuated vaccine against avian pneumovirus in turkeys

Since 1997, avian pneumovirus (APV) has caused estimated annual losses of $15 million to the Minnesota turkey industry. In order to develop an attenuated live vaccine against APV, we serially passaged a Minnesota isolate of APV (APV/MN/turkey/1-a/97) in vitro in cell cultures for 41 passages. Laboratory experiments with this high-passage virus (P41) indicated that the attenuated virus provided immunogenic protection to turkeys against challenge with virulent APV, although some birds showed mild to moderate dinical signs after inoculation. To reduce the residual pathogenicity of P41, while maintaining its immunogenicity, we decided to vaccinate turkeys with P41 in the presence of an immunomodulator, S-28828 (1-n-butyl-2-ethoxymethyl-1H-imidazo[4,5-c]quinolin-4-amine-hydrochloride), which is a potent cytokine inducer. The combined inoculation of S-28828 (5 mg/kg body weight) and P41 resulted in a significant reduction in the incidence of virus-induced clinical signs in comparison with birds that received P41 without immunomodulator (P < 0.05). Only 17% of birds inoculated with S-28828 + APV P41 showed mild respiratory symptoms at 5 days postinoculation as compared with 46% of the vaccinated turkeys that did not receive S-28828. Vaccination with either P41 or with P41 + S-28828 protected turkeys against dinical signs and viral replication after challenge with virulent APV. These results indicate that immunomodulators, such as S-28828, may act as good vaccine adjuvants that can reduce the pathogenicity but maintain the immunogenicity of partially attenuated vaccines.     

Assessment of clonal fidelity of micropropagated gerbera plants by ISSR markers

True-to-type clonal fidelity is one of the most important pre-requisites in micropropagation of crop species. Genetic fidelity of in vitro raised 45 plants of gerbera (Gerbera jamesonii Bolus) derived from three different explants, viz., capitulum, leaf and shoot tips, was assessed by 32 ISSR markers, for their genetic stability. Out of 32 ISSR markers, 15 markers produced clear, distinct and scorable bands with an average of 5.47 bands per marker. The markers designed from AG motif amplified more number of bands. The markers anchored at 3′ ends produced high number of consistent bands than unanchored markers. Fifteen ISSR markers generated a total of 3773 bands, out of which 3770 were monomorphic among all the clones. The Jaccard's similarity coefficient revealed that out of 45 clones derived from different explants, 44 were grouped into a single large cluster alongwith the mother plant with a similarity coefficient value of 1.00, whereas one clone (C38) remained ungrouped. The clones derived from capitulum and shoot tip explants did not show any genetic variation, whereas, one of the leaf-derived clones exhibited some degree of variation.     

Race-specific genetics of resistance to black rot disease [Xanthomonas campestris pv. campestris (Xcc) (Pammel) Dowson] and the development of three random amplified polymorphic DNA markers in cauliflower

Summary

In order to understand the genetics of resistance to black rot disease caused by Xanthomonas campestris pv. campestris (Xcc) (Pammel) Dowson in cauliflower (Brassica oleracea var. botrytis L.) and to identify random amplified polymorphic DNA (RAPD) markers that segregate with the resistance genes, susceptible (‘Pusa Himjyoti’, female parent) and resistant (‘BR-161’, pollen parent) plants were crossed. Six generations of plants (30 P₁, 30 P₂,30 F₁, 120 F₂, 90 B₁, and 90 B₂) were evaluated for the presence or absence of black rot disease in a randomised block design with three replications. The pattern of segregation of resistance was tested by the χ² test at the 5% level of significance. All F₁ progeny plants were resistant, and the segregation of resistant and susceptible plants in the F₂ and two backcross generations (B₁ and B₂) showed that a single dominant gene caused resistance to the black rot pathogen in ‘BR-161’. Three polymorphic RAPD markers (OPO-04₈₃₃, OPAW-20₂₅₃₈, and OPG-25₆₂₅) were found by bulk segregant analysis, which produced unique amplicons 833 bp, 2,538 bp, and 625 bp in length, respectively. These markers were associated in coupling phase to the resistance allele. Best fit ratios of 3:1 (resistant:susceptible) in the F₂ plants with the three RAPD markers, suggested that the markers were linked to the single gene controlling black rot resistance. These markers will be useful to identify more closely-linked markers and to develop black rot-resistant hybrid cauliflower varieties.     

Jasmonic and salicylic acid-induced resistance in sorghum against the stem borer Chilo partellus

Induced resistance was studied in three sorghum genotypes (IS2205, ICSV1 and ICSV700) against Chilo partellus (Swinhoe) (Lepidoptera; Pyralidae) infestation and jasmonic acid (JA) and salicylic acid (SA) application. The activity of plant defensive enzymes [peroxidase (POD), polyphenol oxidase (PPO), superoxide dismutase (SOD), and catalase (CAT)], and the amounts of total phenols, hydrogen peroxide (H₂O₂), malondialdehyde (MDA), and proteins were recorded at 6 days after infestation. The induction of enzyme activities and the amounts of secondary metabolites varied among the genotypes and treatments. The genotype IS2205 showed a stronger effect than that of ICSV1 or ICSV 700. Treatment with JA followed by insect infestation induced greater levels of enzymes and secondary metabolites. The results suggest that JA induces greater levels of resistance components in sorghum plants against insect pests. Thus, pretreatment of plants with elicitors including JA and SA could provide a greater opportunity for plant defense against herbivores.     

Suppression of ovarian progesterone production in dairy cows using an implant of GnRH-agonist (deslorelin) for the purpose of evaluating progesterone metabolism

OBJECTIVE: To evaluate the potential of an implant of a GnRH-agonist (deslorelin) to create a progesterone free animal suitable for studying progesterone (P4) metabolism in intact cows by measuring blood P4 and faecal P4 metabolites. METHODS: Experiment 1: Eighteen non-lactating cycling Holstein-Friesian cows, 4 to 7 years old, were allocated to one of three groups to study plasma P4 concentrations preceding an intravaginal insert. These groups comprised: i) a deslorelin group (GnRH-agonist implanted); ii) a PGF group receiving two injections of prostaglandin (PGF2alpha) 12 days apart; and, iii) an ovariectomised (OVX) group. An intravaginal device (CIDR) was inserted into the vagina of each animal and left in place for 11 days. Plasma P4 concentrations were measured during the study period. Experiment 2: Twelve non-lactating cycling Holstein-Friesian cows, 4 to 7 years old, were allocated to two groups: i) a deslorelin group (GnRH-agonist implanted); and ii) an ovariectomised group. Plasma P4 and faecal P4 metabolites (20-oxo-pregnanes, 20alpha-OH and 20beta-OH) were monitored for a period of 5 weeks. RESULTS: Experiment 1: Average plasma P4 concentration did not differ between the three groups (1.28, 1.43 and 1.55 ng/mL for deslorelin, OVX and PGF cows, respectively, P = 0.8) during the period of supplementation. Experiment 2: There was no difference in plasma P4 (mean plasma P4 < 0.02 ng/mL, P = 0.9) and faecal P4 metabolites between deslorelin and OVX cows 2 weeks after the implantation (P = 0.7). CONCLUSIONS: These data showed that a GnRH-agonist (deslorelin) implant may be used as an alternative to ovariectomy to create a progesterone free animal suitable for studying the metabolism of administered P4.     

Growth-inhibiting activity of azadirachtin onCorcyra cephalonica

The growth-inhibiting properties of azadirachtin, a bioeffective triterpenoid compound isolated from neem (Azadirachia indica A. Juss) seeds, was evaluated on the rice moth,Corcyra cephalonica Staint., a serious pest of stored products. By topical application of various azadirachtin doses (0.5 to 10 μg/larva) in methanol to last-instar spinning stage larvae, development was inhibited. The effect was dose-dependent; at higher doses, many of the insects remained in the larval stage (55% with 10 μg/larva). Disturbance of both larval-pupal and pupal-adult molting is discussed and interpreted as interference with the morphogenetic hormone pool size.     

The effects of bovine respiratory syncytial on normal ovine lymphocyte responses to mitogens or antigens in vitro

In the present study peripheral blod mononuclear cells (MNC) obtained from normal uninfected lambs were used to study the possible effects of bovine respiratory syncytial virus (BRSV) on lymphocyte responses to the mitogens, phytohaemagglutinin (PHA), concanavalin A (Con A) and pokeweed mitogen (PWM) in vitro. Live BRSV had a depressive effect on the proliferative responses of normal MNC to PHA, Con A and PWM. Inactivated BRSV and a commercial preparation of prostaglandin E2 were also found to depress the proliferative responses of normal ovine MNC to PHA but recombinant tumour necrosis factor-alpha (TNF-alpha) had no such effect. Serum samples obtained from BRSV-infected lambs contained substances inhibitory to PHA-driven lymphocyte blastogenesis. Memory blastogenic responses to border disease virus (BDV) of lymyphocytes obtained from lambs previously primed with BDV were significantly reduced when lymphocytes were exposed to infectious BRSV.     

Comparative pathogenesis of serotype 1 and variant serotype 1 isolates of infectious bursal disease virus and their effect on humoral and cellular immune competence of specific-pathogen-free chickens

Specific-pathogen-free chickens inoculated with isolate VA (variant A) or isolate IM of infectious bursal disease virus (IBDV) were examined for mitogenic response to T-cell mitogens, primary and secondary antibody response to sheep erythrocytes and Brucella abortus, and gross and histologic lesions in thymus and bursa. Both isolates induced comparable depression in the mitogenic and antibody response, and both caused extensive gross and histologic lesions in the bursa of Fabricius. However, bursal necrosis induced by the IM isolate was accompanied by an inflammatory response, whereas the inflammatory component was lacking in the lesion induced by the VA isolate. Furthermore, the IM isolate induced extensive lesions in the thymus, but the VA isolate did not.     

Evaluation of Cytotoxic Responses Caused by Selected Organophosphorus Esters in Chick Sympathetic Ganglia Cultures

Ten day old chick sympathetic ganglia cultured in a microslide assembly were treated with a selected group of organophosphate pesticides to evaluate their cytotoxicity ranges, and the usefulness of such a model for screening pesticides. Examination by phase contrast and light microscopy for chemically-induced morphological alteration of nerve fibers, glial cells and neurons provided the criteria for quantitation and assessment of the toxic effects. Concentrations that produced half-maximal effects ranged from 1 × 10^-6M (severely toxic) for methylparathian, diazinon, paraoxon, mevinphos, diisopropylfluorophosphate, tri-o-tolyl phosphate and its mixed isomers to a 1 × 10^-3M (intermediate) for malathion, leptophos, coumaphos, mono- and dicrotophos. Some or no effects were evident at 1 × 10^2-M for O'ethyl-O-p-nitrophenyl phenyl phosphonothioate, tri-m-tolylphosphate, chlorpyriphos and triphenyl phosphate. In all instances, nerve fibers were more sensitive than neurons or glial cells to insecticides. All cellular growth was inhibited at 1 × 10^-2M (except triphenyl phosphate). Below 1 x 10^-7M, no inhibitory effects were evident. The secondary abnormalities included decreased cellular migration, diffuse cellular growth pattern, increased vacuolization, nerve fiber swelling and cellular degeneration. The cytotoxic effects of these chemicals do not appear to be related to in vivo toxicity or cholinesterase inhibition potential.