Detection and phylogenetic characterization of astroviruses in insectivorous bats from Central‐Southern Italy

In recent years, bats have been found to harbour many viruses, raising several questions about their role as reservoirs and potential disseminators of zoonotic viruses. We investigated the presence of six virus families in bats in three regions of Central‐Southern Italy. Astroviruses were identified in seven of 13 bat species. Sequence analysis revealed marked genetic heterogeneity among the astroviruses identified, with nucleotide identity ranging between 60.26% and 87.62%. Astrovirus diversity was not associated with the bat species, the geographic areas or the bat colony, suggesting the circulation of several astrovirus strains in Italian ecosystems. Genetic diversification and interspecies transmission appear common in bat astroviruses and could provide, potentially, the bases for transmission to humans and other mammals. Yet overemphasizing this risk might have detrimental consequences for bat conservation and preservation of the important ecosystem services bats provide.     

Experimental infection of dogs with a novel strain of canine coronavirus causing systemic disease and lymphopenia

A pantropic canine coronavirus (CCoV) strain (CB/05) has been recently associated to a fatal outbreak of systemic disease in young dogs. We report the clinical, virological and serological findings in dogs experimentally infected with strain CB/05. The dogs, three 2.5-month-old and two 6-month-old pups, were successfully infected, shedding viral RNA with their faeces for the entire observation period (21 days) and displaying systemic clinical signs resembling those observed during the course of natural infection. Leucopenia (acute lymphopenia) occurred in all infected dogs, with values dropping below 60% of the initial counts. Considering the severity of the CB/05-induced disease, two of the youngest pups were euthanized for ethical reasons at days 8-9 postinfection, whereas the other pups underwent a slow but progressive improvement of their clinical status with complete recovery. At postmortem examination, remarkable lesions were observed in the internal organs of the euthanized pups, that tested positive for CCoV by real-time RT-PCR and virus isolation on cell cultures. All pups seroconverted for CCoV, as shown by the high optical density values and antibody titres detected by ELISA and virusneutralisation tests, respectively. The present study confirms that strain CB/05 is highly pathogenic for dogs, being able to induce a severe disease (and in some cases the death) even in experimental conditions.     

Screening techniques and sources of resistance to nematodes in cool season food legumes

Identification of sources of resistance in cool season legumes to cyst (Heterodera spp.), root-knot (Meloidogyne spp.), and stem nematode (Ditylenchus dipsaci) is generally based on number of cysts on roots, root-knot nematode induced gall index, and stem nematode reproduction in shoot tissue, respectively. Various levels of resistance to cyst nematodes have been detected in chickpea and pea. Resistance has also been identified in chickpea, faba bean, and pea to the root-knot nematodes. Broad based durable sources of resistance to plant parasitic nematodes are required. Basic research is needed to develop transgenic plants with resistance based on hatch stimulants, inhibitors, toxins, or repellents found in antagonistic rhizosphere microorganisms. Selection of genotypes that favor development of beneficial rhizosphere microorganisms or root endophytes that increase the plant resistance to nematode infection deserves attention.     

Reversing agriculture from intensive to sustainable improves soil quality in a semiarid South Italian soil

Intensive agriculture (IA) is widespread in South Italy, although it requires frequent tillage, large amounts of fertilizers and irrigation water. We have assessed the efficacy of reversing IA to sustainable agriculture (SA) in recovering quality of a typical South Italy soil (Lithic Haploxeralf). This reversion, lasting from 2000 to 2007, replaced 75% of nutrients formerly supplied inorganically by farmyard manuring and reduced the tillage frequency. Several chemical and biochemical properties, functionally related to C and N mineralisation–immobilisation processes and to P and S nutrient cycles, were monitored annually from 2005 to 2007 in the spring. Reversing IA to SA decreased soil bulk density, almost doubled the soil organic matter (SOM) as favoured the immobilisation of C and N, increased most soil microbial indicators but decreased contents of nitrate, mineral N and K₂SO₄-extractable C. The K₂SO₄-extractable C/K₂SO₄-extractable organic N ratio suggested that substrate quality rather than the mass of readily available C and N affected biomass and activity of soil microflora. Also, the largely higher 10-day-evolved CO₂–C-to-inorganic N ratio under SA than IA indicated that higher C mineralisation, associated with higher microbial biomass N immobilisation, occurred under SA than IA. Decreases in most soil enzyme activities under IA, compared to SA, were much higher than concomitant decreases in SOM content. Soil salinity and sodicity were always higher in IA than SA soil, although not critically high, likely due to the intensive inorganic fertilisation as irrigation waters were qualitatively and quantitatively the same between the two soils. Thus, we suggest that the cumulative small but long-term saline (osmotic) and sodic (dispersing) effects in IA soil decreased the microbial variables more than total organic C and increased soil bulk density.     

Environmental factors that affect the ability of amylose to contribute to retrogradation in gels made from rice flour

Retrogradation in rice is a trait that describes the hardening of cooked rice after storage or cooling, and it has significant implications for many consumers of rice, since many people cook rice in the morning and consume it several hours later or the next day. Tools to select against retrogradation in breeding programs are yet to be described. Here, we aim to determine the effect on retrogradation of storage time and temperature and the role of starch, protein, and lipids using gels made from Koshihikari grown in either Australia or Japan. Immediately after cooking, cooling from 60 to 40 degrees C had a minimal effect on firmness, but cooling to 20 degrees C led to significantly firmer gels. Storing the gels at low temperatures did not have an additional effect on the firmness as compared with storing the gels at 20, 40, or 60 degrees C. The removal of proteins led to significantly softer gels at all storage treatments but did not affect the change in firmness on cooling. The removal of lipids increased the rate of retrogradation and the firmness of gels significantly for all treatments. Koshihikari grown in Japan retrograded much less than Koshihikari grown in Australia. The amount of amylose that could be washed from gels made from Australian flour was much greater than for gels made from Japanese flour. After storage, only low molecular weight amylose chains were released from the gel and only after rewarming them to 60 degrees C. Despite the fact that flours from both origins were 18% amylose, the amount of long amylose chains that were complexed with lipids was much greater for the Japanese rice, and the unavailability of the complexed long amylose chains explained the lack of retrogradation in the Japanese rice. Once the long chains were released from amylose-lipid complexes, the Japanese rice retrograded. Thus, the environmental factor affecting retrogradation in this variety is type or amount of lipids synthesized, and the degree of retrogradation was determined by the availability of long chains of amylose.     

Differential cell counts in canine cytocentrifuged bronchoalveolar lavage fluid: a study on reliable enumeration of each cell type

Background: Bronchoalveolar lavage (BAL) allows cell recovery from the lower respiratory tract; differential cell counts of BAL fluid gives important information in the assessment of various bronchial and pulmonary diseases. To the best of our knowledge no study has investigated the relation between the number of cells counted and the reproducibility of BAL fluid differential cell counts. Objective: The purpose of this study was to investigate using statistical methods how many cells should be counted in cytocentrifuged BAL fluid preparations in order to obtain a reliable enumeration of each cell type. Methods: BAL fluid samples from dogs with suspected bronchopulmonary disease were obtained during fiberoptic bronchoscopy with a standardized protocol. Differential cell counts were performed on May–Grünwald–Giemsa‐stained cytocentrifuged preparations by 2 independent observers. Reproducibility for the enumeration of each cell type was expressed as the intraclass correlation coefficient. We considered a threshold level of ≥0.90 to be high and a threshold level of ≥0.85 to be adequate. Results: Forty BAL fluid samples were included in the study. For neutrophils, alveolar macrophages, and eosinophils high reproducibility was reached by counting 200 cells; adequate reproducibility was reached for lymphocytes and bronchial epithelial cells by counting 500 cells. Conclusions: A 500‐cell differential count is required for all types of cells to be quantified with adequate reproducibility in canine cytocentrifuged BAL fluid samples.     

Low temperature storage of pistachio pollen

Summary Pollen from four male pistachio (Pistacia vera L.) clones was stored at –196°C and –20°C for up to 12 months and tested for ability to germinate in vitro following a period of hydration at high humidity. Germination of fresh pollen was high (>80%) for each clone. At –196°C, pollen of cv. Peters survived freezing, storage and thawing with no loss of germinability; pollen of the other three clones had sharp declines in germination possibly attributable to cellular lesions incurred during freezing or thawing. When the relative humidity of the –20°C storage environment was maintained at or near 33%, Peters pollen had high rate of germination through 12 months storage. Without control of relative humidity, Peters pollen germination was high at 4 months, but declined at 12 months. Germination requirements became more exacting for pollen stored at –20°C for 12 months at suboptimal humidity conditions. Pollen of the other three clones did not tolerate storage at –20°C as well as Peters pollen regardless of the storage humidity environment.     

Welfare assessment in intensive and semi-intensive dairy cattle management system in Sicily

The present study aimed to compare the welfare of dairy cows kept in two traditional husbandry systems (semi-intensive and intensive farming) in south-eastern Sicily. A total of 18 dairy farms (nine semi-intensive and nine intensive) were evaluated with a multicriteria system adapted for Sicilian conditions and obtained simplifying the model of the European Food Safety Authority (EFSA). Values of welfare measures, collected by inspections of the farms (general well-being indicators, ventilation system, resting areas [cubicles or bedding], flooring, milking parlours and waiting area, manger and watering equipment), and those of health categories (cases of abortions, hypocalcemia, displacement of abomasum, acidosis/ketosis, enteritis, hoof problems, and mastitis) obtained through the farm records, were compared using Mann–Whitney and Chi-squared tests, respectively. Data showed significant differences (p ≤ .05) about the variables related to welfare categories such as housing ventilation system, resting area, manger, and water equipment that were better in the semi-intensive system than the intensive system. No significant differences were observed about the variables related to health indicators. The results demonstrated that in Sicily the semi-intensive farm is better than the intensive to satisfy the conditions of animal welfare.     

Full-length genome analysis of G2, G9 and G11 porcine group A rotaviruses

Group A rotaviruses with G2 and G9 VP7 specificity are common in humans, while G11 strains have been detected only sporadically. G2, G9 and G11 rotaviruses also circulate in pigs and swine rotaviruses have been suspected of interspecies and zoonotic transmissions in numerous studies. However, the complete gene constellation of G2 and G9 porcine rotaviruses has not yet been determined. In order to start filling this gap, the genomic make up of two G2, one G9 and one G11 porcine rotavirus strains, detected in Canada in 2005–2007, was determined. With the exception of a G2P[34] strain, with E9 NSP4 type and mixed I5 + I14 VP6 type, the constellation of genomic segments was rather conserved and were closely related to prototype porcine strains in the four viruses characterized (I5-R1-C1-M1-A8-N1-T7-E1-H1). Most notably, all the viruses displayed a rare NSP3 genotype, T7, which has also been identified in rare human reassortant strains and in the reference strain RVA/Cow-tc/GBR/UK/1973/G6P[5]. This study provides crucial genetic data on these complex viruses and will help understand the origin and ecological niche of gene segments and the role played by pigs in their evolution.