IHHN Virus as an Etiological Factor in Runt-Deformity Syndrome (RDS) of Juvenile Penaeus vannamei Cultured in Hawaii

Runtdeformity syndrome (RDS) is an economically significant, frequent disease problem of cultured Penaeus vannamei . RDS is characterized by variable, often greatly reduced, growth rate of up to 30% of a cultured population and many shrimp with cuticle deformities of the rostrum, anterior appendages or other parts. The cause of RDS is undetermined.
Nursery trials comparing histologically IHHN-positive and histologically IHHN-negative Penaeus vannamei cultured under identical conditions were conducted at The Oceanic Institute. The IHHN-positive populations developed RDS symptoms including increased individual size variation, reduced population growth, and high prevalance of rostrum, antennae or cuticle deformity, while the IHHN-negative groups displayed none of these symptoms. No other diseases or parasites were identified in the IHHN-positive populations that would account for the RDS symptoms.
A single commercialele nursery was stocked with histologidly IHHN-positive P. vannamei and a high prevalence of RDS was observed. Shrimp growth was bimodal with a subpopulation growing normally and a subpopulation growing slowly. Significant relationships between shrimp harvest size and IHHN prevalance, selected individual organ IHHN severity grades and IHHN severity index (six organs evaluated) were found. Large, apparently normal shrimp were less severely infected with virus and did not display any cuticle deformities.
In sum, these data provide epidemiological and histopathological evidence for the hypothesis that infection by IHHN virus is the cause of RDS in cultured P. vannamei .     

Habitat correlates of distribution and local population decline of the endemic Sardinian newt Euproctus platycephalus

Habitat-based statistical models relating patterns of presence and absence of species to habitat variables could be useful to resolve conservation-related problems and highlight the causes of population declines. In this paper, we apply such a modelling approach to an endemic amphibian, the Sardinian mountain newt Euproctus platycephalus, considered by IUCN a critically endangered species. Sardinian newts inhabit freshwater habitat in streams, small lakes and pools on the island of Sardinia (Italy). Reported declines of newt populations are not yet supported by quantitative data, however, they are perceived or suspected across the species' historical range. This study represents a first attempt trying to statistically relate habitat characteristics to Sardinian newt occurrence and persistence. Linear regression analysis revealed that newts are more likely to be found in sites with colder water temperature, less riparian vegetation and, marginally, absence of fish. The implications of the results for the conservation of the species are discussed, and suggestions for the short-term management of newt inhabited sites suggested.     

Simple assay for analyzing five microcystins and nodularin in fish muscle tissue: hot water extraction followed by liquid chromatography-tandem mass spectrometry

A simple, specific, and sensitive procedure for determining six cyanotoxins, that is, microcystins RR, LR, YR, LA, and LW and nodularin, in fish muscle tissue is presented. This method is based on the matrix solid-phase dispersion technique with heated water as extractant followed by liquid chromatography (LC)-tandem mass spectrometry (MS) equipped with an electrospray ion source. Target compounds were extracted from tissue by 4 mL of water acidified to pH 2 and heated at 80 degrees C. After acidification and filtration, 0.2 mL of the aqueous extract was injected in the LC column. MS data acquisition was performed in the multireaction monitoring mode, with at least two precursor ion > product ion transitions selected for each target compound. Analyte recovery ranged between 61 and 82% and was not substantially affected by either the analyte concentrations or the type of fish. The nonexcellent recovery of some of the microcystins was traced to binding of these compounds to protein phosphatases in fish tissue occurring during sample treatment. The existence of covalently bound microcystins in fish has been evidenced by several studies. Compared to an older sample preparation procedure, this one extracted larger amounts of the analytes in a simpler and much more rapid way. On the basis of a signal-to-noise ratio of 10, limits of quantification were estimated to range between 1.6 and 4.0 ng/g. The effects of temperature and volume of the extractant on the analyte recovery were studied.     

Comparison between auricular and standard rectal thermometers for the measurement of body temperature in dogs

Although the rectal mucosa remains the traditional site for measuring body temperature in dogs, an increasing number of clinicians have been using auricular temperature to estimate core body temperature. In this study, 88 mature healthy dogs had body temperatures measured with auricular and rectal thermometers. The mean temperature and confidence intervals were similar for each method, but Bland-Altman plots showed high biases and limits of agreement unacceptable for clinical purposes. The results indicate that auricular and rectal temperatures should not be interpreted interchangeably.     

Rate of apoptosis in feline mammary tumors is not predictive of postsurgical survival.

Invasion, cell proliferation and apoptosis are important biological features of neoplasia, bearing prognostic importance. Histological stage, mitotic index, and apoptotic index have been assessed in 33 feline malignant mammary tumors. Histological stage (P < 0.01) and mitotic index (P < 0.001) had a significant association with prognosis in univariate analysis. Apoptotic index did not correlate with survival (P = 0.44), and histological stage (P = 0.48) did not correlate with mitotic index (P = 0.39). In feline malignant mammary tumors the apoptotic index seems unable to predict survival and lacks any correlation with proliferation assessed as mitotic index. A possible explanation for the lack of correlation between apoptotic index and survival may be due to the rapid acquisition of pathways of apoptosis resistance in feline mammary tumors or to rapid hormone receptors loss.     

Endoscopic bronchial anatomy in the cat

The aim of this paper is to define the endoscopic bronchial anatomy of the cat and to adapt these findings to the bronchoscopic nomenclature previously described for the dog by Amis and McKiernan (Amis TC, McKiernan BC (1986) Systematic identification of endobronchial anatomy during bronchoscopy in the dog. American Journal of Veterinary Research 47(12), 2649-2657). In this study, the normal endoscopic bronchial anatomy was identified based on analysis of 10 air-dried feline lungs, two euthanased and eight healthy anesthetized cats using either a rigid or a flexible endoscope. Endoscopic photographs were taken during the examinations in the anesthetized cats for a photographic atlas. Corrosion casts were made from the 10 air-dried lungs to confirm the anatomy identified by bronchoscopy. With the results of our findings a bronchoscopic map of the feline bronchial tree and a photographic bronchoscopic atlas of the healthy cat were made.     

Population genetic diversity of the endemic Sardinian newt Euproctus platycephalus: implications for conservation

The Sardinian mountain newt Euproctus platycephalus, endemic to the island of Sardinia, (Italy), is considered a rare and threatened species and is classed as critically endangered by IUCN. It inhabits streams, small lakes and pools on the main mountain systems of the island. Threats from climatic and anthropogenic factors have raised concerns for the long-term survival of newt populations on the island. MtDNA sequencing was used to investigate the genetic population structure and phylogeography of this endemic species. Patterns of genetic variation were assessed by sequencing the complete Dloop region and part of the 12SrRNA, from 74 individuals representing four different populations. Analyses of molecular variance suggest that populations are significantly differentiated, and the distribution of haplotypes across the island shows strong geographical structuring. However, phylogenetic analyses also suggest that the Sardinian population consists of two distinct mtDNA groups, which may reflect ancient isolation and expansion events. Population structure, evolutionary history of the species and implications for the conservation of newt populations are discussed.     

Columns and Detectors Recommended in Gas Chromatography to Measure Greenhouse Emission and O2 Uptake in Soil: A Review

ABSTRACT

Gas chromatography (GC) is a technique used to analyze substances/molecules (as chemical species) with a system composed of chromatograph, column, and detector. This study has the objective of reviewing the use of GC in monitoring greenhouse gases (GHG; carbon dioxide: CO₂; methane: CH₄; nitrous oxide: N₂O) emission and O₂ (oxygen) uptake in soil, demonstrating results from experiments around the world and alternative use of sensors to monitor these gases in soil. Our study shows that the correct column and detector depend on analyzing gas and the advantages and disadvantages of the column and the detector. The columns, packed and capillary, have been more used and are considered better options to analyze GHG emission and O₂ uptake in soil science. Thermal conductivity detector (TCD), electron capture detector (ECD) and mass selective (MS) are great choice to monitor CO₂ emission; flame ionization detector (FID) equipped with methanizer allows the detection of CO₂ and CH₄ emission; and ECD detects the amount of N₂O emitted. Moreover, both, TCD and ECD, also can be used to detect O₂ uptake. GC system is complex, and to identify of GHG emission and O₂ uptake is necessary the use of column (packed or capillary) and may contain multiple detectors, i.e. three (TCD, ECD, and FID) or two detectors (FID and ECD). Field and laboratory study should be run to verify the efficacy of sensors to monitor GHG emission and O₂ uptake as an alternative of GC system.     

Mast seeding in deciduous forests of the northern Apennines (Italy) and its influence on wild boar population dynamics

Context

Pulsed food resources may strongly affect the population dynamics of several consumer species, with consequences on the ecosystem. One of the most common pulsed resources is forest mast seeding.

Aims

We analysed mast seeding in deciduous forests in a mountainous area of northern Apennines and its effect on population dynamics of wild boar (Sus scrofa L.).

Methods

We performed a quantitative, 20-year analysis on annual seed production in Turkey oak (Quercus cerris L.), beech (Fagus sylvatica L.) and chestnut (Castanea sativa Mill.) forest stands using litter traps. The wild boar population density was estimated by means of drive censuses and hunting bag records. The role of other biotic (density of predators) and abiotic (climate) factors potentially affecting wild boar mortality was also investigated.

Results

Turkey oak and chestnut showed high levels of seed production, whereas lower levels were found in beech. The pulsed resources of chestnut and Turkey oak positively affected piglet density. Analyses also highlighted the influence of snow cover and wolves on wild boar population dynamics.

Conclusion

Wild boar can be considered a pulse rate species, the management of which can be improved by annual monitoring of seed production.     

Lymphocyte activation as cytokine gene expression and secretion is related to the porcine reproductive and respiratory syndrome virus (PRRSV) isolate after in vitro homologous and heterologous recall of peripheral blood mononuclear cells (PBMC) from pigs vaccinated and exposed to natural infection

The present study evaluated the lymphocyte activation in PRRSV-vaccinated pigs subsequently exposed to natural infection by in vitro stimulation of peripheral blood mononuclear cells (PBMC) with homologous vaccine and two heterologous PRRSV isolates. The responsiveness was assessed by determining IFN-γ secreting cells by ELISpot assay, lymphocyte CD8 phenotype by intracellular staining/flow cytometry, cytokine gene expression by real-time quantitative PCR and cytokine secretion by ELISA. Conventional pigs were weaned at 28 days of age and inoculated intramuscularly (IM) or needle-less intradermally (ID) with a modified-live PRRSV vaccine suspended in adjuvant, while control pigs were injected with adjuvant alone (ADJ). Blood samples were collected at vaccination, 35 days post-vaccination and after 35 days post-exposure to natural infection by a heterologous field strain. Thirty-five days post-vaccination, PRRSV vaccine induced a low but significant virus-specific IFN-γ secreting cell response upon stimulation with both the vaccine strain and the two isolates in vaccinated pigs. Conversely, after 35 days post-exposure, only the vaccine strain and the BS/114/S isolate triggered this response. Intracellular staining showed that PRRSV-specific immune cells reacting upon vaccine strain and BS/114/S stimulation were mostly CD8⁺ IFN-γ producing cells whereas the stimulation with BS/55 isolate induced an IFN-γ production associated to the CD8^?IFN-γ⁺ phenotype. At 35 days post-vaccination, PBMC from vaccinated pigs showed lower IL-10 expression and release, and higher TNF-α gene expression upon stimulation with both the vaccine and viral isolates. After infection, both cytokines were not differently modulated in different groups. Immune parameters give evidence that IFN-γ secreting cells in the peripheral blood can be elicited upon PRRSV infection although vaccination itself does not stimulate high levels of these reactive cells. Moreover, the cross-reactivity against divergent PRRS viruses can show a different intensity and be differently associated with cytotoxic CD8⁺IFN-γ⁺ as well as CD8^?IFN-γ⁺ cells. Overall, the obtained data confirmed that the immune activation against PRRSV is not dependent on the genetic divergence of the virus. Especially after infection, a different immune reactivity was evident upon stimulation with the different isolates in terms of frequency and CD8 phenotype of PRRSV-specific IFN-γ producing cells. The modulation of cytokines in vaccinated pigs appeared to be more dependent on vaccination or infection conditions than on stimulation by different isolates, and the changes of IL-10 more relevant than those of TNF-α at gene and protein levels. Moreover, under the conditions of this study, the PRRSV vaccine administered via the intradermal route by a needle-less device was confirmed to induce an immune response comparable or in some cases higher than the intramuscular route.