Accuracy assessment and screening of a dairy herd with paratuberculosis by three different ELISAs

Although the culture of Mycobacterium avium subsp. paratuberculosisis is the gold standard for the diagnosis of paratuberculosis, this bacterium is difficult to grow. In contrast, serological tests like ELISAs are inexpensive, rapid, and easy to perform. The aims of this study were to evaluate the accuracy of three different ELISAs: one with the commercial antigen PPA-3, another one with L5P (a recently described lipopentapeptide), and a third one with an in-house antigen whole cell lysates (WCL) of M. avium (MAA) strain D4ER (Study 1), and to compare them with other tests for paratuberculosis (PTB) diagnosis (Study 2). In Study 1, the sensitivities of the three ELISAs tested were 74.1%, 37% and 74.1%, respectively, whereas their specificities were 98.9%, 100% and 100%, respectively. In Study 2, we compared the three above-mentioned ELISAs with the intradermal reaction test using Avian PPD (PPDa) and fecal culture associated with Ziehl-Neelsen stain and PCR tests, in a dairy herd with 4.6% of cows with clinical signs of PTB. The results showed that fecal samples from 14 cows (16%) were culture-positive and that fecal samples from nine cows (10%) were PPDa-positive. Most of these animals (culture-positive and PPDa-positive) were detected as positive with any of the three ELISAs tested. Serological results showed that 31% of the animals were positive to ELISA-PPA-3, 17% to ELISA-L5P and 42.5% to ELISA-WCL. The combination of these three ELISAs identified 50.6% of the animals as positive in the infected herd. In particular, the results show that the locally developed ELISA seems to be useful for identifying many infected animals in a herd.     

Partially defatted black soldier fly larva meal inclusion in piglet diets: effects on the growth performance,nutrient digestibility,blood profile,gut morphology and histological features

Background: The aim of this trial was to investigate the effects of different inclusion levels of a partial y defatted black soldier fly(BSF, Hermetia il ucens L.) larva meal on the growth performance, nutrient digestibility, blood profile, gut morphology and histological features of piglets. A total of 48 newly weaned piglets were individual y weighed(initial body weight(IBW): 6.1 ± 0.16 kg) and randomly al ocated to 3 dietary treatments(4 boxes as replicates/treatment and 4 animals/box). BSF larva meal was included at increasing levels(0% [BSF0], 5% [BSF5] and 10% [BSF10]) in isonitrogenous and isoenergetic diets formulated for two feeding phases: I(from d 1 to d 23) and I(from d 24 to d 61). The weight gain(WG), average daily gain(ADG), average daily feed intake(ADFI) and feed conversion ratio(FCR) were calculated for each feeding phase and for the whole trial. The haematochemical parameters and nutrient digestibility of the piglets were also evaluated. A total of 3 piglets per box were slaughtered on d 61 and the slaughtered piglets were submitted to morphometric investigations and histopathological examinations.Results: No overal significant differences were observed for growth performance(P 0.05), except for the ADFI of phase I, which showed a linear response to increasing BSF meal levels(P 0.05, maximum for the BSF10 group). Dietary BSF meal inclusion did not significantly influence the blood profile, except as far as monocytes and neutrophils are concerned,and these showed a linear and quadratic response, respectively, to increasing BSF meal levels(P 0.05, maximum for the BSF10 and BSF5 groups, respectively). On the other hand, the nutrient digestibility, gut morphology and histological features were not affected by dietary BSF meal inclusion(P 0.05).Conclusions: The obtained results show that a partial y defatted BSF larva meal can be used as a feed ingredient in diets for weaned piglets without negatively affecting their growth performance, nutrient digestibility, blood profile, gut morphology or histological features.     

Laparoscopic ovariectomy in dogs: comparison between laparoendoscopic single-site and three-portal access

This study was conducted to evaluate the feasibility and therapeutic safety of laparoendoscopic single-site ovariectomy (LESS-OVE) and 3-portal laparoscopic ovariectomy (Lap-OVE) in dogs. Ten female mixed breed dogs were included in the study. Dogs were divided into group 1 (LESS-OVE; n = 5) and group 2 (Lap-OVE; n = 5). All procedures were performed by laparoscopic-skilled surgeons, and the anesthetic protocol was the same for all patients. In both groups, the ovarian vascular pedicle and ligaments were transected using a bipolar vessel sealer/divider device. The mean total surgical time was slightly longer in LESS-OVE (36.6 ± 3.5 min) than Lap-OVE (32.0 ± 3.0 min); however, the differences were not significant. Perioperative complications were not reported in any group. Both laparoscopic techniques were shown to be equally feasible and safe for patients. However, surgeons found LESS-OVE to require more skill than Lap-OVE. Therefore, additional studies should be conducted to evaluate this novel approach in clinical veterinary practice, and a proper laparoscopic training program for veterinary surgeons should be developed.     

The effect of land use change and ecotourism on biodiversity: a case study of Manuel Antonio,Costa Rica,from 1985 to 2008

Development in biodiversity rich areas is of global concern. While development may lead to socioeconomic benefits, this often comes concomitant with biodiversity loss and deforestation. Biodiversity rich areas present the opportunity for both improvements in socioeconomic conditions and conservation; however numerous challenges exist. Costa Rica’s Manuel Antonio National Park presents an ideal case study to investigate the balance between alternative forms of development which have contrasting environmental impacts. The Manuel Antonio region is a highly dynamic landscape experiencing deforestation, from agriculture, cattle ranching and oil palm plantations; and also reforestation from abandonment of land holdings and nature oriented tourism. Landscape dynamics are closely intertwined with the livelihoods and perspectives on biodiversity conservation of local communities, determining ecological sustainability. We use an analysis combining multi-temporal remote sensing of land cover dynamics from 1985 to 2008 with questionnaire data from local families on their socioeconomic status, perspectives on conservation, and perceived changes in local wildlife populations. Our results show that, while regeneration occurred and forest fragmentation in the area decreased from 1985 to 2008, Manuel Antonio National Park is rapidly becoming isolated. Decreasing ecological connectivity is related to the rapid expansion of oil palm plantations adjacent to the park and throughout the lowland areas. Perceived decreases in wildlife abundance and compositional change are evident throughout the area, with local communities attributing this primarily to illegal hunting activities. Nature based tourism in the area presents an effective strategy for conservation, including reductions in hunting, through increased valuation of biodiversity and protected areas, and socioeconomic advantages. However, without urgent efforts to limit deforestation and preserve the remaining forested corridor connecting the park to core primary forest, the ability to maintain biodiversity in the park will be reduced.     

Current status of commercial Calibrachoa cultivars as assessed by morphology and other traits

To study the morphological diversity of the popular ornamental crop Calibrachoa, traded since the 1990s, floral and vegetative characters of 91 commercial cultivars were compared to those of natural species. Floral colours were classified according to the ISCC-NBS system and the CIE LAB system. The numerical data obtained by measuring 19 floral and vegetative characters was tested with principal component (PC) analysis. Floral colour classes of commercial cultivars were divided into 37 groups according the ISCC-NBS system, whereas those of natural species were divided into 9 groups. These tendencies were readily observable in the scatter diagrams of the a* vs. b* values and the C* vs. L* values. As the result of PC analysis, six PCs were obtained. For PC4, which negatively correlated with the degree of nocturnal corolla limb closure, the majority of natural species showed negative scores whereas the majority of cultivars had positive PC4 scores. In terms of PC2 (flower size), PC3 (growth form), PC5 (floral shape) and PC6 (leaf shape), natural species were dispersed in a range wider than the cultivars. These results suggest that Calibrachoa cultivars possess more highly variable floral colour than natural species, and breeding programmes to diversify the floral colour seem to have progressed rapidly. The tendency of natural species to close the corolla limb toward evening has been removed almost completely from the cultivars. The floral size, growth form, floral shape and leaf shape of the cultivars were rather uniform compared to those of natural species.     

High temperatures and time to budbreak in low chill apricot ‘Palsteyn’. Towards a better understanding of chill and heat requirements fulfilment

The efficiency of different temperature cycles in inducing budburst of one-year-old shoots of the apricot cultivar ‘Palsteyn’ from dormancy was evaluated. Three replications of shoots were collected during two consecutive years from adult trees, following the accumulation of different amounts of chilling in the field. Thereafter, shoots were exposed to different temperature cycles in growth chambers, for 60 days. The temperature treatments included a continuous temperature of 5 °C; daily temperature cycles of 19/5 h at 5/15 °C, at 5/20 °C, and at 5/25 °C; and the same temperature cycles for the remainder of the 60-day period, after pretreatment at 5 °C for 30 or 45 days. After the temperature treatments, shoots were forced at 25 °C until budburst. The mean time to budburst (MTB) (in days) of lateral vegetative, terminal vegetative and reproductive buds was evaluated. The efficiency of the different treatments was greatly influenced by the date on which shoots were cut. High temperatures had a more positive effect on the reduction of MTB when chilling accumulation had occurred in the field instead of the growth chamber. After partial chilling accumulation in the field, high temperatures (25 °C) combined with low temperatures are more efficient than cycles of moderate temperatures (15 or 20 °C) to induce an earlier budburst. In view of these results, a parallel accumulation of both chilling and heat requirements after partial chilling accumulation is suggested. The application of these results could assist in the development of more accurate models for the prediction of the overcoming of dormancy and blooming.     

A novel system for gene silencing using siRNAs in rice leaf and stem-derived protoplasts

Background  

Transient assays using protoplasts are ideal for processing large quantities of genetic data coming out of hi-throughput assays. Previously, protoplasts have routinely been prepared from dicot tissue or cell suspension cultures and yet a good system for rice protoplast isolation and manipulation is lacking.