Prevalence of Francisella noatunensis subsp. orientalis in Cultured Tilapia on the Island of Oahu,Hawaii

Abstract

Francisellosis is an emergent disease in cultured and wild aquatic animals. The causative agent, Francisella noatunensis subsp. orientalis (Fno), is a gram-negative bacterium recognized as one of the most virulent pathogens of warmwater fish. The main objective of this project was to investigate the prevalence of Fno in cultured tilapia (specifically, Mozambique Tilapia Oreochromis mossambicus, Koilapia [also known as Wami Tilapia] O. hornorum, Blue Tilapia O. aureus, and Nile Tilapia O. niloticus hybrids) on the island of Oahu, Hawaii, using conventional and real-time PCR assays followed by statistical modeling to compare the different diagnostic methods and identify potential risk factors. During 2010 and 2012, 827 fish were collected from different geographical locations throughout the island of Oahu. Upon collection of fish, the water temperature in the rearing system and the length of individual fish were measured. Extraction of DNA from different tissues collected aseptically during necropsy served as a template for molecular diagnosis. High correlation between both molecular methods was observed. Moreover, the bacterium was isolated from infected tilapia on selective media and confirmed to be Fno utilizing a species-specific Taqman-based real-time PCR assay. Although a direct comparison of the prevalence of Fno between the different geographical areas was not possible, the results indicate a high prevalence of Fno DNA in cultured tilapia throughout the farm sites located on Oahu. Of the different tilapia species and hybrids currently cultured in Hawaii, Mozambique Tilapia were more susceptible to infection than Koilapia. Water temperature in the rearing systems and fish size also had a strong effect on the predicted level of infection, with fish held at lower temperatures and smaller fish being more susceptible to piscine francisellosis.

Received November 30, 2012; accepted February 22, 2013     

Effect of vaccination of cattle with the low virulence Nc-Spain 1H isolate of Neospora caninum against a heterologous challenge in early and mid-gestation

Live vaccines have emerged as one of the most potentially cost-effective measures for the control of bovine neosporosis. Previous studies have shown that Nc-Spain 1H is a naturally attenuated isolate of Neospora caninum and that immunisation with live Nc-Spain 1H tachyzoites generated a protective immune response in mice. The aim of this study was to evaluate the safety and efficacy of immunisation in cattle. N. caninum-seronegative heifers were immunised subcutaneously twice with 10⁷ live Nc-Spain 1H tachyzoites prior to artificial insemination. No adverse reactions or negative effects on reproductive parameters were recorded following immunisation. In immunised and non-challenged heifers, no foetal deaths were observed, and none of the calves was congenitally infected. The efficacy against N. caninum-associated foetal death and vertical transmission was determined after challenge with high doses of the Nc-1 isolate at 70 and 135 days of gestation, respectively. After the challenge in early gestation, the immunisation induced a protection of 50% against foetal death. In addition, the microsatellite analysis performed in PCR-positive tissue samples from foetuses that died after challenge infection showed that the profiles corresponded to the challenge isolate Nc-1. A degree of protection against vertical transmission was observed after challenge at mid-gestation; calves from immunised heifers showed significantly lower pre-colostral Neospora-specific antibody titres than calves from the non-immunised/challenge group (P < 0.05). Strong antibody and interferon gamma responses were induced in the immunised heifers. This study indicates that the immunisation before pregnancy with the Nc-Spain 1H vaccine isolate appeared to be safe and reduced the occurrence of N. caninum-associated abortion and vertical transmission in experimentally infected cattle. In light of these encouraging results, the next step for testing this live attenuated candidate should be the assessment of its efficacy and safety in naturally infected cattle.     

Genetic characterization of a novel bovine papillomavirus member of the Deltapapillomavirus genus

This report describes the complete genomic sequence and taxonomic position of BPV type 13. The BPV13 genome was amplified using the multiply primed rolling-circle amplification technique and long-template PCR employing two specific primers. The two long PCR fragments obtained were cloned and sequenced via primer walking. The complete genomic sequence of the BPV13 contains 7961 bp encoding eight proteins, E1, E2, E4, E5, E6, E7, L1, and L2. Similarly to the E5 gene in BPVs 1 and 2, the putative BPV13 E5 ORF encodes a small transforming protein that contains a hydrophobic transmembrane domain. Meanwhile, the retinoblastoma tumor suppressor-binding domain is absent in the putative BPV13 E7 protein. The presence of these two specific molecular features has been recognized as a distinct marker for the development of fibropapilloma in artiodactyl PV-induced lesions. The phylogenetic analysis demonstrated that BPV13 is a new member of the Deltapapillomavirus genus, to be classified as the third representative of the Delta 4 species. The characterization of the genomic sequence of this novel PV will aid in the interpretation of the pathologies described to be related to this virus and provide support for the development of diagnostic tools for epidemiological surveillance of BPV13 in its potential natural hosts.     

Understanding Mediterranean Pasture Dynamics: General Tree Cover vs. Specific Effects of Individual Trees

The study investigated the effect of general and homogeneous tree cover on grassland composition on an extensive Mediterranean rangeland with sparse oak trees in central Spain. We analyzed this effect together with other significant factors identified in this type of rangeland: topography and plowing. Data were collected in the 1984 growing season and they form part of a historical database on the characteristics of vegetation and livestock behavior; these data refer to grasslands below and away from the tree crowns of 91 individual trees, located in different topographical positions and in areas that were last plowed at different times. We used multivariate analyses to identify the main compositional trends of variation in pasture communities. The results indicate that the herbaceous community below tree crowns was more similar to that of the lowland areas than to the nearby areas away from the tree. This result supports the idea of tree cover in semiarid rangelands as a factor attenuating the effects on pastures of environmental conditions typical of high and intermediate topographical positions—generally presenting low soil moisture and fertility. Coupled with this, we also found effects of some individual trees related with the way livestock uses them as shelter and resting places. Our results indicate that the role played by dispersed trees in the management of this type of rangeland should be analyzed at two complementary spatial scales: the overall effect of tree cover as a factor acting at landscape scale and the specific effect of some individual trees acting at a more detailed scale.     

Expansion under hypoxic conditions enhances the chondrogenic potential of equine bone marrow-derived mesenchymal stem cells

Bone marrow-derived mesenchymal stem cells (BM-MSCs) are widely used in regenerative medicine in horses. Most of the molecular characterisations of BM-MSCs have been made at 20% O₂, a higher oxygen level than the one surrounding the cells inside the bone marrow. The present work compares the lifespan and the tri-lineage potential of equine BM-MSCs expanded in normoxia (20% O₂) and hypoxia (5% O₂). No significant differences were found in long-term cultures for osteogenesis and adipogenesis between normoxic and hypoxic expanded BM-MSCs. An up-regulation of the chondrogenesis-related genes (COL2A1, ACAN, LUM, BGL, and COMP) and an increase of the extracellular sulphated glycosaminoglycan content were found in cells that were expanded under hypoxia. These results suggest that the expansion of BM-MSCs in hypoxic conditions enhances chondrogenesis in equine BM-MSCs.     

Expression of connexin 26 and bovine papillomavirus E5 in cutaneous fibropapillomas of cattle

Bovine papillomaviruses (BPVs) can infect epithelial cells and fibroblasts, inducing fibropapillomas in cattle. Gap junctions are communication channels between cells composed of connexins (Cxs). This study evaluated expression of Cx26 and the major BPV oncoprotein E5 in bovine cutaneous fibropapillomas. BPV DNA was amplified from 20/20 fibropapillomas and 3/3 samples of normal skin. All fibropapillomas (20/20) were positive by immunostaining for E5, whereas the three normal skin samples were negative. Cx26 was expressed faintly in the normal skin epithelium. Positive cytoplasmic and juxtanuclear immunoreactivity for Cx26 was evident in 18/20 (90%) fibropapillomas. Western blot analysis demonstrated higher expression of Cx26 in 6/6 fibropapillomas compared to normal skin samples.     

Effects of a weak DC electric field on root growth in Arundo donax (Poaceae)

Abstract

Electric fields can determine changes at the morphological and the physiological level in plants. Plants of Arundo donax L. (giant reed), obtained by set, grown on organic substrate were exposed to a DC electric field. A significant increase in growth rate was observed in the shoots and roots of treated plants. Treated roots also showed a modification in their morphology as compared to untreated ones. Our results point to a possibility of applying electric fields in plant propagation and reproduction.     

Variation of Soil and Water Volumes in Riparian Jungle Soils of Brazil

The objective of this work is to study the relation between humidity, density, porosity and shrinkage of the floodplain soil and riparian vegetation and their ability to store water. For this purpose, two locations for every type of soils were evaluated. Both were placed at the Agronomy University （Faculdade de Cidncias Agron6micas） in SAo Manuel, State of SAo Paulo, Brazil. The floodplain soil was vegetated with Southern Cattail （Typha domingensis）. In both places, soil samples were collected from several depths： 0, 30, 60 and 100 cm. Results show that lower soil density values （0.15 g/cm3） with organic texture and high porosities values （up to 86.2%） were found in samples with the highest organic material content in the floodplain soil. For this field experiment, flood plains soils （characterised as basin gley soils） presented high volumetric instability with a retratibility of 67.49% and higher water storage capacities compared to riparian stands soils （characterised as fluvic neosoils）.