Localization of a stable neural correlate of associative memory

Do learning and retrieval of a memory activate the same neurons? Does the number of reactivated neurons correlate with memory strength? We developed a transgenic mouse that enables the long-lasting genetic tagging of c-fos-active neurons. We found neurons in the basolateral amygdala that are activated during Pavlovian fear conditioning and are reactivated during memory retrieval. The number of reactivated neurons correlated positively with the behavioral expression of the fear memory, indicating a stable neural correlate of associative memory. The ability to manipulate these neurons genetically should allow a more precise dissection of the molecular mechanisms of memory encoding within a distributed neuronal network.     

Induced sero-conversion in heifers with a field strain of bovine pestivirus–a comparison of methods and doses

Losses from pestivirus infection in a closed herd of cattle occurred over several years. In order to prevent further losses, controlled exposure of non-pregnant heifers to pestivirus from viraemic carrier animals was undertaken. Two initial experiments were conducted using either intra-nasal EDTA blood or field contact. Subsequently, other yearling heifers were inoculated with various dilutions of serum using subcutaneous, conjunctival and intra-nasal routes. Effective doses were determined. Neither inoculation nor contact infection produced any clinical illness. The highest dilutions of serum at which sero-conversion occurred were conjunctival, undiluted; intranasal, 10(-1) and subcutaneous 10(-5). With the subcutaneous route all heifers sero-converted at 10(-3). The results for the subcutaneous inoculations showed that the 50% infectious dose for cattle was not distinguishable from that determined in cell culture. Inoculation with a field strain of pestivirus in freeze-thawed serum has effectively and safely induced sero-conversion in heifers. Inoculation of all cattle at risk is considered necessary because no secondary transmission from inoculated heifers was observed.     

Spine-type-specific recruitment of newly synthesized AMPA receptors with learning

The stabilization of long-term memories requires de novo protein synthesis. How can proteins, synthesized in the soma, act on specific synapses that participate in a given memory? We studied the dynamics of newly synthesized AMPA-type glutamate receptors (AMPARs) induced with learning using transgenic mice expressing the GluR1 subunit fused to green fluorescent protein (GFP-GluR1) under control of the c-fos promoter. We found learning-associated recruitment of newly synthesized GFP-GluR1 selectively to mushroom-type spines in adult hippocampal CA1 neurons 24 hours after fear conditioning. Our results are consistent with a "synaptic tagging" model to allow activated synapses to subsequently capture newly synthesized receptor and also demonstrate a critical functional distinction in the mushroom spines with learning.     

Rainfall and breed effects on the efficacy of ivermectin jetting fluid for the prevention of fly strike and treatment of infestations of lice in long-wooled sheep

SUMMARY An ivermectin-based jetting fluid was assessed for its efficacy in the prevention of blowfly strike and treatment of lice in long-wooled sheep. The ivermectin concentrate was diluted to 0.03 mg/mL and applied via a standard hand jetting technique. In insectary studies, using laboratory-reared Lucilia cuprina, sheep jetted with ivermectin were protected from induced breech and body strikes for 18 weeks after treatment. Merino and Corriedale sheep were equally protected, as were sheep subjected to 25 mm simulated rainfall at 1, 3 or 6 hours after jetting. Naturally occurring infestations of lice, Bovicola (Damalinia) ovis, were successfully treated with ivermectin jetting fluid in Merino and Corriedale sheep using a standard hand jetting technique. In addition, the efficacy of treatment was not affected by simulated rainfall at 1, 3 or 6 hours after treatment.     

Use of a Piezo Drill for Intracytoplasmic Sperm Injection into Cattle Oocytes Activated with Ionomycin Associated with Roscovitine

Intracytoplasmic sperm injection (ICSI) consists of the introduction, by micromanipulation, of a single sperm into the cytoplasm of a mature egg. This technique is particularly advantageous when only a few sperm are available for fertilization, representing an important tool in preserving genetic material, especially from poorly fertile males. The results from ICSI in cattle are very often unsatisfactory and difficult to reproduce. Thus, the goal of this study was to evaluate the effect of the use of a Piezo drill (PD) and oocyte activation with ionomycin + roscovitine (I + R) during ICSI in cattle oocytes. After in vitro maturation (24 h), cumulus complex oocytes were divided into four groups: G1 – the ICSI was performed without the use of a PD and the oocyte was activated with I + R; G2 – the ICSI was performed with the use of the PD and activation with I + R; G3 – the ICSI was performed with the use of the PD, but without activation and G4 – parthenogenetic control, treated with I + R, but without sperm injection. The presumptive zygotes were cultured for 7 days and evaluated on day 3 for cleavage rate and on day 7 for blastocyst formation. Embryo production by standard in vitro fertilization in the laboratory was 78% for cleavage (117/150) and 35% for blastocyst formation (41/150). The cleavage rates obtained in G1, G2 and G4 were similar (66.7%, 71.6% and 66.3%, respectively), demonstrating the beneficial effect of oocyte activation. However, in G3, despite the presence of the sperm and the electric stimulation of a PD, the cleavage rates were significantly lower (17.5%) compared with the groups that used chemical activation, even in the absence of sperm (G4). Despite the beneficial effects of activation, this stimulus alone, or in the absence of the PD, was not sufficient for adequate morulae formation (13.4%, 37.9%, 0.0% and 13.5% for G1, G2, G3 and G4, respectively). Only in G2, when the PD was used followed by artificial activation, blastocysts were obtained (14.7%). These results indicate that cattle oocytes must be activated after ICSI to produce viable embryos.     

Efficacy of ivermectin jetting fluid against strike by some primary and secondary blowflies of sheep

SUMMARY Merino sheep, which were hand jetted with ivermectin jetting fluid, and untreated sheep were challenged with larval implants of Lucilia cuprina, Lucilia sericata, Calliphora noclva and Chrysomya ruflfacles at intervals of about 2 weeks from 6 to 16 weeks after treatment. Both Lucilia species produced strike rates of about 90% in untreated sheep; the respective rates were lower for Chr rufifacies (55%) and C nociva (60%). Strike rates for the treated group were about 17, 11, and 9% for L cuprina, L sericata, and Chr rufifacies, respectively. Only 1 implant site in the treated group was struck by C nociva. Treated sheep had significantly (P < 0.01) longer time to first strike than did untreated ones for each species of fly. L sericata, Chr rufifacies, and C nociva larvae implanted on treated animals had significantly (P < 0.05) longer time to first strike than did L cuprina larvae.     

Loss of Restriction Site DdeI, used for Avian Molecular Sexing, in Oreophasis derbianus

Molecular sexing is a rapid and safe procedure for bird sex determination. Two universal methods based on the amplification of a chromo‐helicase‐DNA‐Binding 1 (CHD) gene region, located in both sexual chromosomes (Z and W), have been established. We found that molecular sexing of Oreophasis derbianus failed by using these two procedures. One of them is based on a restriction site located in CHD1W gene but absent in CHD1Z. The DdeI restriction site, used successfully to determine gender in several bird species, was found to be lost because of nucleotide change in O. derbianus. This change created a new restriction site, NlaIII, that was successfully applied for sexing this endangered bird.     

Efficacy of ivermectin jetting fluid for the curative treatment of blowfly strikes in sheep

SUMMARY A jetting fluid containing 7.5% w/v ivermectin, which was diluted to 0.03 mg/mL in water and applied using a standard hand jetting technique, was assessed for its efficacy in treating blowfly strikes on Merino sheep. Strikes were induced on sheep held under insectary conditions using laboratory-reared Lucilia cuprina. All struck sheep jetted at 28,48 or 72 hours after exposure to flies were successfully treated. All 1st, 2nd and early 3rd instar larvae were killed within 24 hours of treatment.