Pronuclear Embryo Yield in Canindé and Saanen Goats for DNA Microinjection

The objective of this study was to examine the effect of donor breed on pronuclear‐stage embryo yield to be used for DNA microinjection in a transgenesis goat program. Twelve Canindé and twelve Saanen goats were heat synchronized using a progestagen‐cloprostenol treatment. Forty‐eight hours before the sponge removal, superovulation was induced with a total administration of 4.4 mg/kg bodyweight NIH‐FSH‐P1, given twice daily in decreasing doses over 3 days. In addition, goats received 100 μg of GnRH and they were hand‐mated at 36 and 48 h after progestagen removal. Embryo recovery was performed by oviduct flushing at 72 h after sponge removal. Embryos were microinjected with a DNA construct and noticeable swelling of the nuclei was the criterion for successful microinjection. The total diameter, cytoplasm diameter, zona pellucida thickness and pronuclei diameter were measured for each microinjected embryo. A higher (p < 0.05) percentage of fertilized ova was observed in Canindé (89.9%) than Saanen (36.2%) goats. In addition, Canindé donors produced a higher percentage of pronuclear embryos when compared with Saanen: 72.5% vs 20.6% (p < 0.05), respectively. Successful microinjection was verified in 96.7% and 73.3% of times in Canindé and Saanen embryos, respectively (p < 0.05). Significant differences were observed for all morphometric parameters except for cytoplasm diameter. In conclusion, under our study experimental conditions, Canindé were more efficient than Saanen goats concerning the pronuclear embryo yield and manipulation. The use of Canindé goats in transgenesis programs could be increase the interest in their breeding and could be contribute to saving them from extinction.     

Hepatozoon spp.: report of some cases in dogs in Brasília, Brazil

Canine hepatozoonosis is a disease caused by the tick-borne protozoan Hepatozoon spp. It has been reported in the United States, southern Europe, the Middle East, Africa and the Far East. In Brazil, canine hepatozoonosis is an emerging protozoal tick-borne disease, and is characterized by distinct clinical signs. The objective of this study was to analyze the laboratory findings of some hepatozoonosis cases in dogs in Brasília, Brazil, and their clinical signs. The animals of this experiment showed low parasitemia, similar to H. americanum, but the clinical signs presented were similar to H. canis. According to our observations and in agreement with O'Dwyer et al. [Vet. Parasitol. 94 (2001) 143], the Brazilian Hepatozoon appears more to resemble the species found in the eastern Hemisphere than with H. americanum of North America, or could be caused by a new species. Our data revealed that hepatozoonosis could be considered endemic in Brasília.     

Seed dormancy and germination in the summer annual Galeopsis speciosa

This study examined germination and dormancy in Galeopsis speciosa (Lamiaceae), a common summer annual weed in cold‐temperate areas. Seeds collected in southern Sweden were subjected to several experiments. The seeds were dormant at maturity. Seeds sown outdoors after collection produced a small number of seedlings that emerged early in the spring. After long cold stratification or stratification outdoors over two winters, the maximum germination was 40–50%; germination occurring over a wide range of temperatures. Warm stratification preceding cold stratification had no effect on germination, but repeated warm and cold periods seemed to promote germination. Gibberellic acid (GA) stimulated germination, but full germination was only achieved after more than 2 months of incubation at the most suitable temperature regime tested. Excised embryos grew and developed into normal seedlings. With these results, the species does not fit into the currently used system for seed dormancy classifications. The response to GA and the growth of excised embryos indicate non‐deep or intermediate physiological dormancy, but dormancy alleviation by stratification was not in line with the guiding principles for these classifications. Galeopsis speciosa has a strong dormancy that is sufficiently alleviated during the winter to allow germination of only part of a seed batch each year; hence a stepwise germination pattern occurs over a period of several years.     

Nitrogen mineralization in sandy loam soils under an intensive double-cropping forage system with dairy-cattle slurry applications

Nitrogen (N) mineralization and soil mineral N contents were measured at 2-week intervals over a 2-year period (June 1994–May 1996) on two different sites in the North West region of Portugal. The experiment was established in fields, which had for many years been under a double-cropping forage system with maize from May to September and a winter crop (mixture of cereals and Italian ryegrass) during the rest of the year. In addition to N fertilizers, dairy-cattle slurry was applied regularly at the sowing of each crop. On this intensive forage system, quantification of N released from slurry, crop residues and soil organic matter becomes important when better N use efficiency and reduced environmental impact from agricultural practices are required. Net N mineralization rates of the 0–10 cm soil layer fluctuated considerably between consecutive incubation periods and ranged from −0.88 to 1.87 mg N kg⁻¹ day⁻¹ with annual average rates of between 0.41 and 0.65 mg N kg⁻¹ day⁻¹. The total N mineralized in the 10 cm depth soil layer reached values between 122 and 224 kg N ha⁻¹ year⁻¹, showing that mineralization was a very important N source for the crops. The amounts of N released during the cold season (November–February) were equivalent to 27–48% of the yearly total. Regression analysis indicated that seasonal variation in N mineralization was only poorly explained by soil moisture and temperature. The changing balance during the year between soil moisture and temperature will contribute to the relatively constant N mineralization rates. Soil mineral N contents during the maize crop were high and exceeded the nutrient requirements for the optimum yield of this crop. Under the climatic conditions of the region and due to the poor development of the winter crop plants at the time, the mineral N left in the soil after the maize crop and released by mineralization during the cold season is particularly vulnerable to nitrate leaching losses.     

Evaluation of auditory evoked potentials to predict depth of anaesthesia during fentanyl/fluanisone−midazolam anaesthesia in rats

Objectives To assess a method for monitoring depth of anaesthesia using components of middle latency auditory evoked potential (AEP) waveforms during anaesthesia with fentanyl/fluanisone and midazolam. Study design Prospective observational study. Animals Five female Wistar rats weighing between 210 and 250 g. Methods Implanted electrodes were used to record AEPs in animals receiving five doses of anaesthetic. Recordings were made at 5 minutes post‐injection (deep anaesthesia; no pedal withdrawal response, PWR) and then at 25 minutes (light anaesthesia; strong PWR). Responses showed five characteristic peaks occurring at 11, 14, 23, 42 and 68 ms that were measured for latency of occurrence and peak amplitude. Results Auditory evoked potential peaks P14, N23 and P42 were increased significantly in latency with successive anaesthetic injections [avg. F(1,4) = 12.53, p < 0.001; avg. F(1,4) = 10.6, p < 0.001; avg. F(1,4) = 3.9, p = 0.02, respectively]. Peak N23 showed a significant reduction in latency during the 20 minute recovery period following both the first and second anaesthetic injections (t(3) = 7.52, p = 0.005; t(4) = 5.17, p = 0.007, respectively). Peak P42 occurred significantly earlier 20 minutes following the second anaesthetic injection (t(4) = 4.75, p = 0.009). The mean overall depth of anaesthesia assessed using PWR scores was significantly correlated with the mean latency of peak N23, such that as the strength of PWR increased, N23 occurred significantly earlier (r = ?0.99, p = 0.01). The amplitude difference between peaks N23 and P42 increased after the second and third drug administrations [avg. F(1,4) = 10.65, p = 0.031 and avg. F(1,4) = 11.24, p = 0.028, respectively]. Conclusion The characteristics of these peaks, and in particular latency of peak N23, may provide a useful tool for assessing depth of anaesthesia produced by this, and possibly other anaesthetic agents.     

Critical disease components of common bacterial blight to effectively evaluate resistant genotypes of snap bean

Common bacterial blight (CBB) caused by Xanthomonas axonopodis pv. phaseoli (Xap) is the main bacterial disease of snap bean. The present work aimed to select snap bean genotypes that are resistant to CBB based on three components of resistance: area under the disease progress curve, latent period, and lesion diameter on pods (DL). A completely randomized two-factor factorial design with six replications was used to evaluate leaves and pods of 14 snap bean and three common bean genotypes (PI 207262, BAC-6 and A-794) with high resistance to CBB. Two Xap isolates, 1394-98 and 775-90, were used to inoculate leaves and pods. Data were analyzed using nonparametric statistics. Significant differences were observed among the evaluated genotypes for all of the components of resistance, except for DL. The snap bean UENF 1482 demonstrated good performance in two disease resistance components. For this reason, this genotype can be recommended for use in breeding programs that aim to generate snap bean genotypes resistant to Xanthomonas axonopodis pv. phaseoli.     

Sward structure management for a maximum short‐term intake rate in annual ryegrass

This study tested the hypothesis that different sward structures, which were constructed by varying the pre‐ and post‐grazing sward heights of annual ryegrass pasture (Lolium multiflorum Lam.) in southern Brazil, affect the short‐term intake rate (STIR) by dairy cows. Treatments consisted of four sward‐management strategies defined by a combination of two pre‐ (25 and 15 cm) and two post‐grazing sward heights (10 and 5 cm): 15‐05, 15‐10, 25‐05 and 25‐10. A completely randomized block design with four replicates was used for the experimental design. The STIR was determined by the double‐weighing technique. Jaw movements were evaluated using automatic recorders (IGER Behaviour Recorder). The results showed that treatment 25‐10 allowed the animals to collect more herbage with a greater bite mass and thus resulted in a greater STIR. Treatments in which sward height was reduced to 10 cm generally favoured the ingestion process than treatments with a post‐grazing sward height of 5 cm.