Bioactive cinchona alkaloids from Remijia peruviana

Three known Cinchona alkaloids of the quinine type, quinine (1), cupreine (2), cinchonine (3), and the possible artifact cinchonine-HCl (3-HCl), along with two new ones, acetylcupreine (4) and N-ethylquinine (5), have been isolated from the bark of Remijia peruviana (Rubiaceae). Their stereochemical structures were established by high resolution NMR spectroscopy. Alkaloids 2-4 had antifeedant effects on Leptinotarsa decemlineata with varying potencies. Compound 4 was cytotoxic to both insect Sf9 and mammalian CHO cells after 48 h of incubation, while 3-HCl had stronger and selective cytotoxicity to Sf9. Quinine 1 had a moderate to low effect on Trypanosoma cruzi. Tumoral cells were also affected by these alkaloids, with 4 and 3-HCl being the most cytotoxic to all the cell lines tested. Overall, the 8R, 9S configurations, as in 3 and 3-HCl, as well as the C-6'acetylated alkaloid 4, with an 8S, 9R configuration, showed stronger biological effects.     

Historical and contemporary habitat use of sperm whales around the Galápagos Archipelago: Implications for conservation

1. Sperm whales have occupied the waters off the Galápagos Islands, Ecuador, for at least the past 200 years. During the 19^th century, they were the target of intensive whaling that severely depleted the population. In recent times, after commercial whaling ended, sperm whales in the region remain vulnerable to multiple threats, especially potential entanglement in fishing gear, which may hinder their ability to recover from the whaling era.
2. As a highly mobile, long-lived species, long-term analysis of the habitat use of sperm whales is necessary to establish effective conservation and management strategies. Here, contemporary (1985–2014) and historical (1830–1850) sperm whale habitat use off the Galápagos Islands was analysed and contrasted to the extent of the Galápagos Marine Reserve (GMR). Contemporary habitat use and its variability over time were modelled as a function of geographic, oceanographic, and topographic variables using generalized additive models.
3. The fine-scale habitat (<50 km) used by sperm whales was associated with topographic (i.e. depth and slope) and oceanographic characteristics (i.e. relative sea surface temperature and standard deviation of sea surface temperature), but these preferences varied over time.
4. While historical and contemporary data indicate that sperm whale habitat primarily occurred within the boundaries of the GMR, in recent years, whales were found up to 30.1% of the time outside the GMR, potentially overlapping with commercial fisheries operating in the area.
5. The dynamic nature of the relationship of this nomadic species with its habitat highlights the need of large-scale conservation efforts across the Eastern Tropical Pacific region, including the wide-scale enforcement of regulations requiring the use of Automatic Identification System in fishing vessels, the promotion of on-board fisheries observer programmes, the development of adaptive management strategies, and international collaboration to identify and mitigate threats.

     

Genomic characterization of H1N2 swine influenza viruses in Italy

Three subtypes (H1N1, H1N2, and H3N2) are currently diffused worldwide in pigs. The H1N2 subtype was detected for the first time in Italian pigs in 1998. To investigate the genetic characteristics and the molecular evolution of this subtype in Italy, we conducted a phylogenetic analysis of whole genome sequences of 26 strains isolated from 1998 to 2010. Phylogenetic analysis of HA and NA genes showed differences between the older (1998-2003) and the more recent strains (2003-2010). The older isolates were closely related to the established European H1N2 lineage, whereas the more recent isolates possessed a different NA deriving from recent human H3N2 viruses. Two other reassortant H1N2 strains have been detected: A/sw/It/22530/02 has the HA gene that is closely related to H1N1 viruses; A/sw/It/58769/10 is an uncommon strain with an HA that is closely related to H1N1 and an NA similar to H3N2 SIVs. Amino acid analysis revealed interesting features: a deletion of two amino acids (146-147) in the HA gene of the recent isolates and two strains isolated in 1998; the presence of the uncommon aa change (N66S), in the PB1-F2 protein in strains isolated from 2009 to 2010, which is said to have contributed to the increased virulence. These results demonstrate the importance of pigs as mixing vessels for animal and human influenza and show the presence and establishment of reassortant strains involving human viruses in pigs in Italy. These findings also highlighted different genomic characteristics of the NA gene the recent Italian strains compared to circulating European viruses.     

Pasteurella multocida contains multiple immunogenic haemin- and haemoglobin-binding proteins

Iron-dependent outer membrane proteins (IROMPs) play an important role in bacterial pathogenesis and present several attributes of potential vaccine candidates. TBLASTN analysis of the Pasteurella multocida Pm70 genome using the same molecules of other bacterial pathogens as a query identified eight putative haemin and haemoglobin receptors for this organism. Quantitative binding assays have demonstrated that the proteins PM0040, PM0236, PM0741, PM1081, PM1428, PM0592 and HgbA bind both haemin and haemoglobin, whereas PM0576 and PM1282 ORFs only bind either haemoglobin or haemin, respectively. Furthermore, Western blot analysis showed that P. multocida-infected mice generate specific antibodies against PM0040, PM0236, PM0741, PM1081, PM1428, PM0592 and HgbA proteins. Nevertheless, inoculation of mice with any single one of these receptors alone did not protect against P. multocida infection.     

Combined effects of temperature and diet on growth and biochemical composition of the Pacific oyster Crassostrea gigas (Thunberg) spat

Temperature and quality of the available food are important factors that influence the physiology of oysters; however, the combined effects have not been well studied. We evaluated the impacts of the temperature and diet on the growth, survival and biochemical composition in the Pacific oyster Crassostrea gigas spat, cultured in the laboratory for 8 weeks at 23, 26, 29 and 32°C and fed Isochrysis sp.‐Pavlova lutheri (IP) and Dunaliella tertiolecta (Dt). The growth and biochemical composition showed a pattern, which changed in response to rising temperature. The shell length was significantly longer, in spat fed the IP diet, except at 32°C, where both diets produced poor growth results. The survival was <50% after 5 weeks at 32°C, whereas at all other temperatures it was >88%. High temperatures directly increased lipids and saturated fatty acids, while the proteins, carbohydrates and unsaturated fatty acids decreased. High temperatures achieved in the environment, as those reached on clear summer days during low tides, are an important stressor in oyster spat, especially when the quality of the available food is poor.     

Factorial analysis of the effect of sugarbeet vinasses on agroecological indices in horticultural field

The impact on soil of applying sugarbeet vinasses （V） was analyzed through a field experiment in horticulture greenhouse, arranged in a 23 factorial design. Two levels of three independent variables--application of V, use of polyethylene cover （PC） on the soil. and soil depth （D）---on various dependent variables were studied. Vinasses favoured crop yield and reduced the number of Meloidogyne incognita juveniles in soil. The concentrations ofN. P and K increased with the interaction VxD, with PC also increasing N concentration. The amounts of humic acids and humin decreased with D; fulvic acid concentration increased with V, but decreased with the interaction VxPC. Soil physical factors were improved mainly with D and V. Aromaticity of humic acid-like fractions increased ~＇itb V. In general, V showed significant effects mainly on the topsoil, suggesting low leaching risks. The results indicate that the levels of the independent factors improving a group of variables were not the same that those contributing to another group. Therefore, their best combination should be determined for each scenario to achieve optimum agroecological performance.     

Fish larvae: zooplankton relationships in microcosm simulations of earthen nursery ponds. II. brackish water system

A simulation of the effects of predation intensity on zooplankton composition in brackish water nursery ponds was carried out in order to address the problem that commercial fish nurseries encounter in obtaining enough zooplankton of adequate species composition and size when fish larvae start to feed. The experimental system consisted of twelve 130 l containers with treatments of four densities (0, 1, 2, or 4 larvae l⁻¹) of common carp (Cyprinus carpio L.), stocked on the 6th day after filling the containers. Zooplankton-environment relationships were explored using factor analysis. Factor analysis allowed identifying several groups of zooplankters that responded in different ways to fish larvae predation pressure. The first factor represented a general measurement of rotifer abundance, and the second identified the direct effect of size-selective fish predation. Since no rotifers were present in the filling water, all these species were autochthonous populations that hatched from resting forms in the sediment and reproduced. In the absence of fish predation, this led to a steep rotifer increase. Fish predation started when the rotifer concentration was just starting to increase and their direct predation reduced and delayed the rotifer abundance peak. This effect increased with the increase in fish larvae density. Estimations of rotifer consumption by fish larvae in this experiment were higher than similar calculations from data of the literature, which led us to test the hypothesis that factors other than direct predation were affecting rotifer population dynamics. The mechanisms involved in rotifer population regulation are discussed. It was concluded that in commercial nurseries, increased larvae production can be achieved by keeping the larvae density at an intermediate level and stocking fish to match the increasing phase of the rotifer peak. Under reasonable larvae density (up to 2 l⁻¹) it seems that the direct predation effect of fish larvae on rotifer dynamics is minor, compared to fish induced self regulation.     

The use of infrared spectroscopy and machine learning tools for detection of Meloidogyne infestations

Plant parasitic nematodes are generally soilborne pathogens that attack plants and cause economic losses in many crops. The infested plants show nonspecific symptoms or, often, are symptomless; therefore, diagnosis is performed by taking soil and root tissue samples. Here, we show that a combination of different infrared spectra analysis and machine learning algorithms can be used to detect plant parasitic nematode infestations before symptoms become visible, using leaves instead of roots and soil as samples. We found that tomato and guava plants infested with Meloidogyne enterorlobii produced different spectral patterns compared to uninfested plants. Using partial spectra from 1,450 to 900/cm as the "fingerprint region", principal component analyses indicated that after 5 (tomatoes) or 8 weeks (guava), plants with no visible symptoms of infestations were positively diagnosed. To improve the early detection response, we used machine learning modelling. A support vector machine (SVM) was used to obtain more robust, accurate models. The SVM model contained 34 support vectors, 17 for each level. The overall performance of the model was >97% and the total accuracy was significantly higher, demonstrating the absence of chance prediction. The best prediction of infestation was obtained at the second and fourth weeks for tomatoes and guavas, respectively, reducing the diagnostic time by half. The combined application of these techniques reduces the processing time from field to laboratory and shows enormous advantages by avoiding root and soil sampling.     

Dynamics of porcine circovirus type 2 infection in a herd of pigs with postweaning multisystemic wasting syndrome

OBJECTIVE: To determine the pattern of infection for porcine circovirus type 2 (PCV2) in a herd of pigs with postweaning multisystemic wasting syndrome (PMWS). ANIMALS: 29 sows and 250 pigs. PROCEDURE: Blood samples were collected from all 3-, 7-, and 12-week old pigs and 59 pigs at 28 weeks of age. Pigs that died during the study were necropsied. Porcine parvovirus and PCV2 antibodies were assayed. A polymerase chain reaction (PCR) was used to detect PCV2 genome in serum of selected pigs. RESULTS: The PMWS started when pigs were 8 weeks old, with a prevalence of 30% in 8- to 10-week-old pigs. Eighty-three pigs died during the period between 3 and 12 weeks of age. Microscopic lesions consistent with PMWS were observed, and PCV2 nucleic acid was detected (50 of 68 pigs). Antibodies to PCV2 decreased from 3 to 7 weeks of age, increased at 12 weeks of age, and were maintained until 28 weeks of age. One sow had a positive result for PCR of serum. Nine, 37 and 8 pigs had PCV2 genome in serum obtained at 7, 12, and 28 weeks of age, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Infection with PCV2 coincided with severe clinical signs; however, infected 28-week-old pigs did not have evidence of disease. Immunity declined over time in young pigs. A long duration of PCV2 viremia was apparent in a high percentage of infected pigs, which may affect transmission and persistence of the virus in a herd.