Genetic engineering of plant volatile terpenoids: effects on a herbivore,a predator and a parasitoid

BACKGROUND: Most insect‐resistant transgenic crops employ toxins to control pests. A novel approach is to enhance the effectiveness of natural enemies by genetic engineering of the biosynthesis of volatile organic compounds (VOCs). Before the commercialisation of such transgenic plants can be pursued, detailed fundamental studies of their effects on herbivores and their natural enemies are necessary. The linalool/nerolidol synthase gene FaNES1 was constitutively expressed from strawberry in three Arabidopsis thaliana accessions, and the behaviour of the aphid Brevicoryne brassicae L., the parasitoid Diaeretiella rapae McIntosh and the predator Episyrphus balteatus de Geer was studied. RESULTS: Transgenic FaNES1‐expressing plants emitted (E)‐nerolidol and larger amounts of (E)‐DMNT and linalool. Brevicoryne brassicae was repelled by the transgenic lines of two of the accessions, whereas its performance was not affected. Diaeretiella rapae preferred aphid‐infested transgenic plants over aphid‐infested wild‐type plants for two of the accessions. In contrast, female E. balteatus predators did not differentiate between aphid‐infested transgenic or wild‐type plants. CONCLUSION: The results indicate that the genetic engineering of plants to modify their emission of VOCs holds considerable promise for facilitating biological control of herbivores. Validation for crop plants is a necessary next step to assess the usefulness of modified volatile emission in integrated pest management. Copyright © 2012 Society of Chemical Industry     

Distinct Effects of Boar Seminal Plasma Fractions Exhibiting Different Protein Profiles on the Functionality of Highly Diluted Boar Spermatozoa

The aim of this study was to evaluate how different protein profiles of seminal plasma (SP) fractions affect sperm functionality in vitro. Ejaculates from three boars were separated into six fractions. The fractions differed from each other in their sperm content, in their total SP protein content, and their spermadhesin PSP-I/PSP-II and heparin-binding protein (HBP) concentrations. Spermatozoa were mainly recovered in fraction 2 (sperm-rich fraction, >1800 × 10⁶ spermatozoa/ml), whereas the pre-sperm fraction 1 and the post-sperm fractions 4–6 contained low numbers of spermatozoa (<500 × 10⁶/ml). Except in fraction 2, the total SP protein concentration and the concentration of both, spermadhesin PSP-I/PSP-II and the HBPs increased with fraction order. Distinct time-dependent effects were observed on motility characteristics and membrane integrity of highly diluted boar spermatozoa upon incubation with a 10% dilution of the SP from each fraction. The highest sperm viability was recorded after exposure for 5 h to fraction 2, followed by fractions 1 and 3. The percentages of motile spermatozoa also differed significantly among fractions after 5 h of incubation. Spermatozoa incubated with SP of fractions 1–3 showed the highest percentage motility. We conclude that different SP fractions exert distinct effects on the functionality of highly diluted boar spermatozoa. Fractions 1–3 appear to promote sperm survival, whereas fractions 4–6 seem to be harmful for preserving the physiological functions of highly diluted boar spermatozoa.     

Effects of Dextrose Plus Lactose in the Sows Diet on Subsequent Reproductive Performance and within Litter Birth Weight Variation

Effects of dextrose plus lactose in sow’s feed were tested on subsequent reproductive performance and within litter birth weight variation. During the last week of gestation and lactation, sows were either fed a commercial lactation diet (Control: C), or an isocaloric diet containing 25 g/kg dextrose plus 25 g/kg lactose (Treatment: T). In the subsequent weaning‐to‐oestrus interval (WEI), all sows received the same amount of a commercial feed, but T sows were supplemented with 150 g dextrose plus 150 g lactose per day. Weight and backfat changes were recorded as well as litter characteristics during the treatment period and the subsequent parity. No significant effect of treatment was found on the subsequent reproductive performance, including the number of piglets born, although the number of live born piglets was 0.51 larger (p = 0.31) and weight of the live born piglets was 84 g higher in the T sows (p = 0.07) than in the C sows. When sows were categorized in sows with 12 or less and more than 12 total born piglets in the previous litter, treatment of sows with dextrose plus lactose resulted for the group with 12 or less piglets in a strong increase in subsequent total born piglets (13.97), whereas in the untreated sows the subsequent litter size was 11.89. In the group with more than 12 total born piglets, no effect of treatment was found (interaction between previous litter size and treatment p = 0.03). The within litter variation in birth weight in the subsequent litter was numerically lower in the T sows. We concluded that the use of dextrose and lactose during lactation and WEI seems to enhance litter size in sows with low previous litter size and seems to have the potential to reduce the within litter variation in birth weight.     

Nutritional evaluation of waste date fruit as partial substitute for soybean meal in practical diets of juvenile Nile tilapia, Oreochromis niloticus L.

The potential of waste date meal (WDM; low-quality date palm, Phoenix dactylifera L.) as a carbohydrate source in formulated diets for Nile tilapia was evaluated. Four isocaloric-practical diets (15.7 kJ g⁻¹) were formulated incorporating WDM at 0, 100, 200 and 300 g kg⁻¹ levels as partial substitutes for soybean meal (SBM). These were designated D₀ [284 g crude protein (CP) and 383 g carbohydrate (CHO) kg⁻¹ diet], D₁ (279 g CP and 446 g CHO kg⁻¹ diet), D₂ (207 g CP and 495 g CHO kg⁻¹ diet) and D₃ (175 g CP and 578 g CHO kg⁻¹ diet). Each diet was fed to three replicate groups of 30 fish [20.20 ± 0.09 g (±SE)] for 75 days. No feed-related mortality was observed during the entire experimental period. Final body weight (FBW) and specific growth rate (SGR) in the different treatments were statistically not significantly different ( P  > 0.05). Protein efficiency rate (PER) was lowest in diet D₀ and increased with decrease of SBM content (D₁–D₃). A significant increase in whole body lipid content was recorded in fish fed diets D₂ and D₃. Results showed that WDM could be a substitute for SBM up to 300 g kg⁻¹ in practical Nile tilapia diets without compromising growth.     

Identification of Polymorphisms in the Osteopontin Gene and Their Associations with Certain Semen Production Traits of Water Buffaloes in the Brazilian Amazon

The osteopontin gene may influence the fertility of water buffaloes because it is a protein present in sperm. The aim of this work was to identify polymorphisms in this gene and associate them with fertility parameters of animals kept under extensive grazing. A total of 306 male buffaloes older than 18 months, from two farms, one in the state of Amapá and the other in the state of Pará, Brazil were used in the study. Seven SNPs were identified in the regions studied. The polymorphisms were in gene positions 1478, 1513 and 1611 in the region 5′upstrem and positions 6690, 6737, 6925 and 6952 in the region amplified in intron 5. The SNPs were associated with the traits, namely scrotal circumference, scrotal volume, sperm motility, sperm concentration and sperm pathology. There were significant SNPs (p < 0.05) for all the traits. The SNP 6690 was significant for scrotal circumference, sperm concentration, sperm motility and sperm pathology and the SNP 6737 for scrotal volume. The genotype AA of SNP 6690 presented the highest averages for scrotal circumference, sperm concentration and motility and the lowest total number of sperm pathologies. For the scrotal volume trait, the animals with the largest volume were correlated with the presence of the genotype GG of SNP 6737. These results indicate a significance of the osteopontin gene as it seems to exert a substantial influence on the semen production traits of male buffaloes.     

Production of Oocytes of Nile Tilapia (Oreochromis niloticus) for In Vitro Fertilization via Hormonal Treatments

Only a few studies have described hormonal treatments for induction of synchronicity and gamete collection in Nile tilapia (Oreochromis niloticus), both important for assortative matings in breeding programmes and essential for polyploidy technologies. In this study, we compared the effectiveness of carp pituitary extract (CPE), Nile tilapia pituitary extract (TPE), human chorionic gonadotropin (hCG) and gonadotropin‐releasing hormone (GnRH) protocols on the induction of spawning and egg production in Nile tilapia. Among the hormonal treatments analysed, only hCG was effective for producing viable gametes for in vitro fertilization. To verify the viability of this hormonal treatment, hCG was tested using different doses (1000, 2000, 3000, 4000 and 5000 IU/kg) in a large number of females (208 animals) from two Nile tilapia lines. The results indicated that hCG doses between 1000 and 5000 IU/kg could be used to induce final oocyte maturation in Nile tilapia with collection of stripped oocytes. This is the first study to report differential reproductive responses to hormonal treatment between tilapia lines: line 1 was more efficient at producing eggs and post‐hatching larvae after hCG induction than line 2. In conclusion, we demonstrated that the hCG protocol may be applied on a large scale to induce final oocyte maturation in Nile tilapia. The development of a protocol for in vitro fertilization in Nile tilapia may aid in breeding programmes and biotechnological assays for the development of genetically modified lines of Nile tilapia.     

An Investigation of Uterine Nitric Oxide Production in Mares Susceptible and Resistant to Persistent Breeding‐Induced Endometritis and the Effects of Immunomodulation

The first objective of this study was to evaluate intrauterine nitric oxide (NO) and endometrial inducible NO synthase (iNOS) in mares susceptible or resistant to persistent breeding‐induced endometritis (PBIE) within 24 h after breeding. Mares susceptible (n = 6) or resistant (n = 6) to PBIE were inseminated over five cycles, and uterine secretions and endometrial biopsies were collected before and 2, 6, 12 and 24 h after insemination. Uterine secretions were analysed for NO and biopsies were analyzed for iNOS expression. A second experiment evaluated the effect of treatment with dexamethasone or mycobacterial cell wall extract (MCWE) on uterine NO production and endometrial iNOS mRNA expression. Six susceptible mares were inseminated over three cycles with (i) killed spermatozoa without treatment (control), (ii) killed spermatozoa with 50 mg of dexamethasone IV or (iii) MCWE IV 24 h prior to insemination with killed spermatozoa. Six resistant mares were inseminated with killed spermatozoa as a control. Six hours after breeding, uterine biopsies and secretions were collected and evaluated for NO and iNOS mRNA. In Experiment 1, resistant mares had an increase in iNOS mRNA expression 2 h post‐breeding compared to baseline (p = 0.045), 12 h (p = 0.014) and 24 h (p = 0.001). Susceptible mares had higher expression 2 h compared to 6 h (p = 0.046). No differences were observed in mRNA or protein expression of iNOS between resistant and susceptible mares. Resistant mares had a relatively steady amount of total intrauterine NO over 24 h, while susceptible mares had an increase over time, with a significantly higher increase in total NO than resistant mares at 6 (p = 0.04) and 12 h (p = 0.032). In Experiment 2, no differences were observed for iNOS mRNA expression. Susceptible mares had increased NO when compared to resistant mares (p = 0.008) and MCWE decreased NO (p = 0.047).     

Some features of the antibody response of sheep

Six nine-month-old red deer were injected intramuscularly with a long-acting injectable solution of oxytetracycline (“Terramycin-LA”, Pfizer Ltd) at a dose rate of 20 mg/kg. Four similar control deer were injected with saline. There was no significant pain response to injection, and only minor palpable swellings at the injection site were observed in three oxytetracycline-treated and one control animal. No statistically significant changes in white blood cell numbers, blood fibrinogen, creatine Phosphokinase or glutamic oxaloacetic transaminase concentrations occurred as a result of Oxytetracycline administration up to seven days after injection. Mean plasma Oxytetracycline concentration reached a peak (4.68 mg/1) two hours after injection and declined to levels below assay sensitivity (0.3 mg/1) in five deer 72 hours after injection, and in all deer by 96 hours after injection. No gross lesions at the injection site were observed at slaughter 30 days after injection. There were traces of Oxytetracycline at the injection site muscle of two deer after 30 days, but residues were not detected in injection site muscle from the other four deer, or in any of the liver, kidney or other muscle specimens.     

Failure to Induce Contact Hypersensitivity in the Opossum,Ittrichosurus Vulpecula

Abstract

Sir:- Contact hyypersensitivity (CHS; syn: contact allergy, contact dermatitis, contact eczema) is induced when chemicals (haptens) capable of binding covalently to autologous proteins are applied to the skin.(1) Claman,, H. N.,, Miller,, S. D.,, Conlon,, P. J. and Moorehead,, J. W. 1980. Advances in Immunology, Edited by: Dixon and, F. J. and Kunkel, H. G. 121–157. New York: Academic Press.  [Google Scholar]The hapten-protein complexes formed in this way are immunogenic and elicit the immune response underlying CHS. CHS is a form of delayed hypersensitivity. Exposure of a primed animal to the sensitising hapten results in erythema and induration at the site of skin contact, the response being maxima1 after 24 hours.