Aujeszky's disease virus infection patterns in European wild boar

Evidence of exposure (i.e. seroprevalence) to Aujeszky's disease virus (ADV) is high among wild boars from south-central Spain. This research aims to determine the presence of ADV by molecular detection, and to describe the patterns of ADV infection in wild boars. Tonsils (TN) and trigeminal ganglia (TG) for ADV molecular detection, and sera were collected from wild boars (n = 192) in 39 hunting estates from south-central Spain (2004/2005). A nested polymerase chain reaction (PCR) for a fragment of the ADV surface glycoprotein B was performed on collected tissues. Individual status of presence of viral DNA was tested against explanatory variables by means of a Generalized Linear Mixed Model (GLIMMIX) analysis. Viral detection prevalence was 30.6 ± 6.7%. Although there was an increasing pattern with age and females presented higher prevalences, no statistically significant influence of sex and age was found for viral presence. Molecular testing in TN and TG allowed classifying infection status into (i) ADV negative (in both TN and TG), (ii) only positive in TN, (iii) only positive in TG and (iv) positive in both TN and TG. ADV DNA was statistically more frequently evidenced in TN in females than in males. With the exception of one individual, all wild boars with presence of ADV DNA in TN and TG or only in TG reacted positive in the ELISA. In contrast, animals with only ADV DNA in TN serorreacted positively and negatively. Interestingly, 45% of the PCR positive wild boars (n = 59) were seronegative in the serological test, all of them with viral DNA only in TN. Our results provide evidence for latency of ADV in wild boars and stress the fact that antibody detection based tests may fail to detect a proportion of recently infected animals. This is of great concern since current management schemes in our study promote animal translocation for hunting purposes, with the associated risk of under-detecting ADV infected individuals when using serology to screen for ADV infection.     

Poor Prediction Value of Sperm Head Morphometry for Fertility and Litter Size in Rabbit

This study was conducted to investigate the predictive capacity of fertility and litter size of sperm head morphometric measurements when the ejaculates fulfilled the minimum requirements commonly used in artificial insemination (AI). Semen samples from 11 rabbits (77 ejaculates) were evaluated for sperm motility, abnormal spermatozoa and sperm head morphometry using computer automated sperm analysis system. Morphometric dimensions for length, width, area and perimeter were analysed. Only ejaculates with more than 70% of motility rate and <15% of abnormal sperm were used for AI. A total of 1031 individual AI were performed in commercial rabbitries. Our results showed significant differences among animals for all sperm head measurements. The mean values for fertility and litter size obtained were 68.4 ± 0.01% and 9.3 ± 0.1% respectively. To assess the predictive value of morphometric dimensions in fertility, a logistic regression analysis was applied. Moreover, multiple linear regression analyses were used to examine the relationship between litter size and sperm head morphometric parameters. Logistic regression analysis rendered a significant model between fertility and area and perimeter, explaining the 0.65% variation. Multiple linear regression analysis rendered a significant model between litter size and width, area and perimeter that explained the 1.3% variation. By conclusion, the sperm head morphometric parameters assay showed low potential to predict fertility and litter size when the ejaculates fulfilled the minimum requirements commonly used in AI (motility and abnormal spermatozoa) in rabbit.     

Effect of a selected Lactobacillus spp.-based probiotic on Salmonella enterica serovar enteritidis-infected broiler chicks

The effect of a Lactobacillus spp.-based probiotic (FM-B11) on Salmonella enterica serovar Enteritidis (SE) recovery was evaluated in liquid (Expt. 1) and lyophilized (Expt. 2) forms in two separate experiments with two trials each. For each trial, 80 broiler chicks were randomly allocated into two treatments: control and probiotic culture. All chicks were challenged with SE (approximately 10(4) colony-forming units [cfu]) upon arrival at our laboratory. In both experiments, probiotic culture was administered in the drinking water for 3 consecutive days at a final concentration of approximately 10(6) cfu/ml, beginning 1 hr after SE challenge. Cecal tonsils were aseptically removed at 24 and 72 hr postchallenge, followed by enrichment and plating on xylose lactose deoxycholate (XLD) agar for the presence or absence of Salmonella-typical colonies. In Expt. 1, a significant reduction (P < 0.05) in SE-positive samples was observed in both trials at 24 and 72 hr postchallenge. Additionally, in Expt. 2, the lyophilized probiotic decreased (P < 0.05) SE recovery at both 24 and 72 hr postchallenge compared with the control group in trial 1. In trial 2, SE evaluation was performed only at 72 hr after challenge and fewer (P < 0.001) treated samples were positive for SE. Results showed that application of either liquid or lyophilized probiotic culture in the drinking water for 3 consecutive days can help to reduce SE recovery from young birds, although further research is needed to elucidate the mechanism of this response.     

Demarcating specific ion (NaCl, Cl, Na) and osmotic effects in the response of two citrus rootstocks to salinity

The present work differentiates the effects of NaCl, Cl⁻, Na⁺ and concentrated macronutrients on two citrus species, sour orange (Citrus aurantium L) and Macrophylla (Citrus macrophylla Wester). Plants were grown in a base nutrient solution (0.07 MPa osmotic pressure) for 4 months before applying the treatments that consisted of isotonic solutions of 0.23 MPa osmotic pressure of Na⁺ (40 mM, without Cl⁻), Cl⁻ (40 mM, without Na⁺), NaCl (40 mM) and 3.5 times the concentration of macronutrients of the base solution. Plants were grown in the different treatment for 2 months before being examined for symptoms of toxicity. The two genotypes showed major differences in the extent of Cl⁻ and Na⁺ accumulation in leaves and in their ability to maintain the internal concentrations of essential nutrients in response to the different ionic compositions of the medium. Differences in mineral nutrient accumulation were observed among treatments in both rootstocks. It was concluded that growth response to the different treatments was primarily affected by an osmotic effect, although in Macrophylla, the ionic effects also seem to be present.     

In vitro establishment of shoot meristems of Ilex paraguariensis and identification of endophytic bacteria

Yerba mate(Ilex paraguariensis A.St.-Hil.)is a species of great economic,social and environmental importance for the southern regions of Brazil,Uruguay and Argentina.Currently the most diverse products are obtained from mate leaves,including mate tea.The objective of this study was to establish shoot meristem cultures(meristematic dome and a few primordia)of elite clones and identify the endophytic bacteria present in the explants.We tested the effect of clones(F1,F2,A03 and A07),culture media(MS,1/2MS,1/4MS and WPM),cytokinins(kinetin,BA and 2iP),activated charcoal(1,2 and 3 g L^-1),and disinfecting agent(sodium hypochlorite and mercuric chloride)on in vitro establishment.F1 and F2 clones were the most responsive for shoot meristem in vitro culture.WPM medium supplemented with 8.8 lM 2iP,0.2 lM NAA and 3 g L^-1 activated charcoal was the most suitable for the in vitro establishment of the F1 clone.No phytotoxic effect of the disinfectant was observed and some meristems sprouted.The isolated endophytic bacterium was identified for the first time in yerba mate as Agrobacterium larrymoorei.To conclude,we were able to establish in vitro culture of yerba mate using meristems as explants but the tissues were not free of endophytic microorganisms which could interfere with explant response.     

Mediterranean fruit fly suppression using chemosterilants for area‐wide integrated pest management

BACKGROUND: The chemosterilisation technique has been demonstrated to reduce the population and fruit damage of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann), in citrus orchards. Field trials showed efficacy by reducing the fruit fly population, which was progressively achieved by continuous application of lufenuron to several generations. Different authors have suggested that field trials should be carried out in isolated or wide areas in order to reduce fruit fly intrusion and obtain best results. To this end, a wide‐area trial over 3600 hectares has been under investigation in Valencia (Spain) since 2002 to validate the chemosterilisation technique against the fruit fly. The whole area was treated with 24 traps ha^?1, using more than 86 000 traps in the field trial. RESULTS: A continuous decrease in fruit fly population was observed over the 4 years under trial. Moreover, results showed a significant reduction in persimmon damage in the chemosterilant treatment area compared with a malathion aerial treatment area. In the case of citrus damage, no significant differences were obtained between malathion and chemosterilant treatments. CONCLUSION: The chemosterilant method reduces Mediterranean fruit fly populations, and therefore it is a candidate treatment to replace aerial treatments with insecticides in order to suppress this pest. In addition, the efficacy of chemosterilant treatment is increasing year after year. The possibility of using this technique combined with other control methods is discussed. Copyright © 2010 Society of Chemical Industry     

Epidemiological study on porcine circovirus type 2 (PCV2) infection in the European wild boar (Sus scrofa)

Porcine circovirus type 2 (PCV2) is considered as the causative agent of postweaning multisystemic wasting syndrome (PMWS) in domestic pigs, where the virus is ubiquitous as evidenced by serological surveys. We present the results of the first nationwide sero-survey on the presence of PCV2 antibodies in European wild boars, and report the first PMWS case in a wild boar from Spain. Sera from 656 hunter harvested wild boars from 45 different geographical sites and 22 additional imported animals were analysed by means of an immunoperoxidase monolayer assay (IPMA). We also examined the tissues from 55 healthy and one diseased wild boars for the presence of PCV2 nucleic acid and PMWS lesions by in situ hybridisation and histopathology, respectively. Additionally, abundance estimates of wild boars and field interviews were carried out on 30 sampling sites. The prevalence of medium to high PCV2 serological titres among the examined wild boars was 47.89 +/- 1.9%. Seropositive wild boars appeared in all but one of the geographical regions analysed. Seroprevalence and titre of PCV2 antibodies were closely related to the management of the wild boar populations. Wild boars from intensively managed, farm-like populations had higher prevalence than wild boars living in more natural situations. The effect of wild boar abundance and management on PCV2 antibody prevalence was further evidenced by the high correlation existing between the relative abundance estimates of animals and the percentage of wild boars with medium to high levels of PCV2 antibodies. PCV2 nucleic acid was detected in the tissues of three wild boars. One of these was diagnosed as PMWS. The results, in addition to information on piglet mortalities, suggest a potential role of PMWS in piglet mortality in intensively managed wild boar populations.     

Contrasting effects of water availability on <Emphasis Type="Italic">Pinus pinaster</Emphasis> radial growth near the transition between the Atlantic and Mediterranean biogeographical regions in NW Spain

Summer water stress is the main limiting factor for Pinus pinaster growth under Mediterranean climate, but no information on limiting physiographic and climatic factors under Atlantic conditions is available for this species. We hypothesize that water availability during the active period limits P. pinaster growth, with stands nearby the Atlantic–Mediterranean boundary being more sensitive to summer drought stress. We analyzed the inter- and intra-annual climatic response of this species at ten monospecific stands in a transitional area between the Atlantic and Mediterranean biogeographical regions in Galicia, NW Spain. Mean sensitivity of radial growth consistently decreased with increasing elevation, and growth variation was quite similar for the earlywood, latewood or total ring widths, suggesting a strong intra-annual coherency of radial growth. Growth patterns were site dependent and geographically structured, with three groups of stands showing different climatic responses. As expected for sites suffering from summer drought stress, water availability enhances growth in western and southern Galicia, showing negative responses to maximum temperatures and positive to water availability. In northern Galicia, away from the Atlantic-Mediterranean boundary, water surplus in the rainy seasons negatively influenced growth. This was probably due to the combined effects of seasonal water-logging stress and the reduction of solar radiation associated with cloudiness, which would limit photosynthetic rates in winter and spring. Local variations of water availability strongly controlled the physiological processes that determine growth dynamics of P. pinaster in NW Spain, contributing to its geographical structure and contrasting sensitivity.     

Location of Prunus Necrotic Ringspot Ilarvirus Within Pollen Grains of Infected Nectarine Trees: Evidence from RT-PCR, Dot-blot and in situ Hybridisation

Prunus necrotic ringspot Ilarvirus (PNRSV) is a pollen and seed-borne ilarvirus affecting most Prunus spp. The location of the virus in infected nectarine pollen grains was investigated by molecular hybridisation, RT-PCR and in situ hybridisation. The first two approaches revealed an internal location of the virus. In situ hybridisation demonstrated the virus in the bicellular pollen grain, where it was present in the cytoplasm of the vegetative cell but not in the generative cell. This result seems to indicate that the sperm cells, formed by the mitosis of the generative cell, are not involved in virus transmission to seed. Other possible mechanisms are discussed.     

Incorporating spatial structure and stochasticity in endangered Bonelli’s eagle’s population models: Implications for conservation and management

Population models have played a chief role informing management decisions for the endangered Bonelli’s eagle (Aquila fasciata) in Spain. In this paper, we incorporate spatial structure and stochasticity in the construction of individual-based metapopulation models, and use these models to explore the effects of possible management actions on the persistence of the species in Spain. To build the models we used data on seven sub-populations that have experienced different trends in the last decades, and we introduced new estimates of pre-adult survival rate. The elasticity analysis of our models showed that when the interchange of individuals among sub-populations is taken into account, pre-adult mortality plays the key role in determining the overall population trend. This is in contrast to what it has been suggested by previous demographic models that modelled local populations as isolated. Specifically, a 20% decrease in pre-adult mortality during the first two years of life was enough for the stabilization of the metapopulation (i.e., λ  1.0). However, neither a similar decrease in the values of adult mortality, nor an increase in the percentage of breeders, modified the declining trend of our model metapopulation. This reinforces the idea that to ensure the long-term persistence of the species in Spain, management actions should aim at minimizing pre-adult mortality. These include locating and protecting the areas used by juvenile Bonelli’s eagles (e.g., temporary settlements), minimizing the risk of electrocution in power lines, and preventing human persecution.