Humoral immune response in pregnant heifers inoculated with Neospora caninum tachyzoites by conjunctival route

The aim of this study was to compare systemic humoral immune responses in pregnant heifers inoculated with Neospora caninum tachyzoites by conjunctival and intravenous routes. Twenty nine heifers separated in three experimental groups were studied: Group 1 (n=10 animals) and Group 2 (n=9 animals) were inoculated with 10(8) of N. caninum tachyzoites by conjunctival and intravenous routes at 5th month of gestation, respectively; Group 3 (n=10 animals) were non-inoculated control animals. An indirect fluorescent antibody test (IFAT) and western immunoblotting (IB) were used to analyze the humoral immune response. All animals from Group 1 developed N. caninum specific antibody responses after conjunctival inoculation recording the highest antibody titer (mean+/-SE: 160+/-49.9) at 6th month of gestation. There were statistical differences between humoral immune responses found in Group 1 and 2 being higher in the second one at 6.5th, 8.5th and 9th months of gestation (P<0.05). Interestingly, all heifers from Group 1 reverted to seronegative status at the end of gestation. No increase in antibody was detected in the uninfected control group. Same pattern of N. caninum antigens was recognized by sera from heifers inoculated by conjunctival route and heifers inoculated by intravenous route. Recognized antigens were 116, 92, 84, 77, 45, 40, 25-26 and 17-18 kDa. The conjunctival instillation of N. caninum tachyzoites in pregnant heifers induces specific systemic antibodies. Further work is needed in order to clarify the consequences of this novel experimental route of infection not only on the fetus but also on the dam.     

Serological screening for Coxiella burnetii infection and related reproductive performance in high producing dairy cows

The possible relationship between Coxiella-seropositivity and the reproductive performance of cows during the previous year to the serological screening was examined in three high producing dairy herds. The herds had a history of subfertility (<25% of pregnancies for the total number of AI), abortion (>18% abortions) and a positive polymerase chain reaction test for Coxiella burnetii in the bulk tank milk with an excretion higher than 10(4)Coxiella /ml for all three herds. Antibodies against C. burnetii were detected in 50.2% of the 781 parous cows analyzed. Coxiella seropositivity was linked to placenta retention, to changes in the interval from parturition to conception (with the lowest interval parturition-conception for cows with low level of seropositivity), early pregnancy (cows becoming pregnant before Day 90 postpartum) and maintenance of gestation during the early fetal period, while it failed to affect rates of abortion after Day 90 of gestation or stillbirth.     

Naphthalophos combinations with benzimidazoles or levamisole as effective anthelmintics for sheep

Objective: To investigate the relative efficacy and safety of the anthelmintic naphthalophos in sheep, either given alone or in combination with benzimidazole (fenbendazole and albendazole) or levamisole anthelmintics.
Design: A parasitological study using faecal egg count reduction tests, a validating slaughter trial and field safety trials.
Procedure: Faecal egg count reduction tests were carried out on 13 farms. Naphthalophos and combinations of naphthalophos with levamisole and fenbendazole were included in the drench tests. On one property a controlled efficacy study was carried out to validate faecal egg count reduction test findings. In this trial, sheep were slaughtered 10 days after treatment and the remaining parasites recovered from the gastro-intestinal tract. Safety trials were carried out on eight farms where approximately 50 000 sheep were treated with naphthalophos and albendazole that were tank mixed in the backpack.
Results: The efficacy of naphthalophos alone in faecal egg count reduction tests ranged from 59 to 98% with one test showing 95% reduction. The efficacy of naphthalophos and levamisole ranged from 74 to 100%, with 5 farms showing 95% reduction. The efficacy of naphthalophos and fenbendazole ranged between 88 and 100% with 95% reduction achieved on 10 farms. The controlled efficacy study showed a good correlation between the faecal egg count reduction tests and numbers of parasites recovered, except for Nematodirus where the faecal egg count reduction tests overestimated efficacy. The mortality rate in the safety trials was 0.05%, with most fatalities occurring on one farm.
Conclusion: The combination of naphthalophos and fenbendazole was more effective than a combination of naphthalophos and levamisole, and will provide a sufficiently safe drench rotation option.     

Genetic diversity and relationships between wild and cultivated olives (<Emphasis Type="Italic">Olea europaea</Emphasis> L.) in Sardinia as assessed by SSR markers

The genetic relationships within and between wild and cultivated olives were examined and clarified in an isolated and restricted area, such as the Mediterranean island of Sardinia. Wild (21 individuals) and cultivated olive trees (22 local cultivars from a germplasm collection and 35 ancient trees) were genotyped by means of 13 SSR loci. Five cases of synonymy were observed and nine distinct genotypes were identified in the collection. Five novel genotypes were also detected among the ancient trees. Differences on the allelic composition and heterozygosity levels were found between wild and cultivated trees. Model-based clustering method classified the olive trees into two major gene pools: (a) wild genotypes and (b) local cultivars from the collection and from heritage olives. Regarding the cultivated plant material, we observed that: (a) most of the Sardinian cultivars shared the same allelic profiles with the ancient cultivated trees and (b) the majority of these cultivars and all the novel genotypes were not related to any other cultivars included in this study. These findings as well as the detection of unique alleles and a certain wild genetic background at some cultivars revealed by the Bayesian analysis may indicate their autochthonous origin. The synonymy cases found between local cultivars and Italian mainland cultivars indicate interchange of genetic material among these growing areas, suggesting thus a possible allochthonous origin. The information obtained can assist in the management of an olive collection and sheds some light on the survival of true oleasters and the origin of Sardinian cultivars.     

Effects of a plastic cover on soil moisture change in a Mediterranean climatic regime

Water deficit is a serious problem for most agricultural crops, especially in arid–semiarid regions, and limits sustainable development. Production can be improved by reducing evapotranspiration and loss of infiltrated water by the use of a plastic cover. We monitored soil moisture fortnightly over 1 yr using a neutron probe near four trees in an olive grove (Olea europaea, var. Arbequina), two of which had a plastic cover buried in the topsoil around them. These trees were monitored using three neutron probe access tubes per tree to compare the behaviour of soil moisture over time with two other identically instrumented and nearby trees with no cover. Analysis was based on the resultant moisture profiles. The plastic cover retained moisture and increased soil water residence time. During the dry season, the amount of water retained in the plots was at most 15–20% greater in the mid‐season and at least 5–6% greater at the end of season than in the central part of the plots near the trunk. The plastic cover was effective to ca. 50 cm with maximum water content near the soil surface. During the wet season, the cover did not affect soil water. Soil moisture was greater near the tree trunk as a result of stemflow and throughfall.     

Flavor release measurement from gum model system

Flavor release from a mint-flavored chewing gum model system was measured by atmospheric pressure chemical ionization mass spectroscopy (APCI-MS) and sensory time-intensity (TI). A data analysis method for handling the individual curves from both methods is presented. The APCI-MS data are ratio-scaled using the signal from acetone in the breath of subjects. Next, APCI-MS and sensory TI curves are smoothed by low-pass filtering. Principal component analysis of the individual curves is used to display graphically the product differentiation by APCI-MS or TI signals. It is shown that differences in gum composition can be measured by both instrumental and sensory techniques, providing comparable information. The peppermint oil level (0.5-2% w/w) in the gum influenced both the retronasal concentration and the perceived peppermint flavor. The sweeteners' (sorbitol or xylitol) effect is less apparent. Sensory adaptation and sensitivity differences of human perception versus APCI-MS detection might explain the divergence between the two dynamic measurement methods.     

Predicting stand damage and tree survival in burned forests in Catalonia (North-East Spain)

The study developed models for predicting the post-fire tree survival in Catalonia. The models are appropriate for forest planning purposes. Two types of models were developed: a stand-level model to predict the degree of damage caused by a forest fire, and tree-level models to predict the probability of a tree to survive a forest fire. The models were based on forest inventory and fire data. The inventory data on forest stands were obtained from the second (1989–1990) and third (2000–2001) Spanish national forest inventories, and the fire data consisted of the perimeters of forest fires larger than 20 ha that occurred in Catalonia between the 2nd and 3rd measurement of the inventory plots. The models were based on easily measurable forest characteristics, and they permit the forest manager to predict the effect of stand structure and species composition on the expected damage. According to the stand level fire damage model, the relative damage decreases when the stand basal area or mean tree diameter increases. Conversely, the relative stand damage increases when there is a large variation in tree size, when the stand is located on a steep slope, and when it is dominated by pine. According to the tree level survival models, trees in stands with a high basal area, a large mean tree size and a small variability in tree diameters have a high survival probability. Large trees in dominant positions have the highest probability of surviving a fire. Another result of the study is the exceptionally good post-fire survival ability of Pinus pinea and Quercus suber.     

Short and long‐term distribution with depth of soil organic carbon and nutrients under traditional and conservation tillage in a Mediterranean environment (southwest Spain)

The impacts of tillage and cropping sequences on soil organic matter and nutrients have been frequently reported to affect the uppermost soil layers, but there is little published information concerning effects at greater depth. This article reports results on the distribution of soil organic carbon (SOC), active carbon (AC), N, Olsen‐P and extractable K within 100 cm in short (4 yr) and long (16 yr) term experiments under different tillage systems. Short (TT₄) and long (TT₁₆) traditional tillage are compared with conservation tillage, reduced (RT₁₆) and non‐tillage (NT₄). The results show more accumulation of SOC in the near‐surface under RT₁₆ and NT₄ in both experiments compared with traditional tillage. Moreover, greater C content occurs to 40 cm depth in the long‐term experiment. The results demonstrate the importance of time on C accumulation, not only in near‐surface layers but also at greater depths. Active C is an indicator of the increase in soil quality in the long‐term experiment. This trend is only apparent for the first 10 cm in the short‐term experiment. Patterns in N, Olsen‐P and extractable K are similar to that of SOC. However, only extractable K is significantly greater in soil under conservation tillage (RT₁₆ and NT₄) after short and long periods. Potassium availability is a good indicator of the changes caused by tillage. Our results indicate that studies of soils at depth could be very useful in long‐term experiments to demonstrate the effect of conservation tillage on C and nutrient distribution.     

Conserved folding in retroviral proteases: crystal structure of a synthetic HIV-1 protease

The rational design of drugs that can inhibit the action of viral proteases depends on obtaining accurate structures of these enzymes. The crystal structure of chemically synthesized HIV-1 protease has been determined at 2.8 angstrom resolution (R factor of 0.184) with the use of a model based on the Rous sarcoma virus protease structure. In this enzymatically active protein, the cysteines were replaced by alpha-amino-n-butyric acid, a nongenetically coded amino acid. This structure, in which all 99 amino acids were located, differs in several important details from that reported previously by others. The interface between the identical subunits forming the active protease dimer is composed of four well-ordered beta strands from both the amino and carboxyl termini and residues 86 to 94 have a helical conformation. The observed arrangement of the dimer interface suggests possible designs for dimerization inhibitors.