Cells infected with Jaagsiekte sheep retrovirus are detected in the bone marrow of asymptomatic sheep

Ovine pulmonary adenocarcinoma (OPA) is a transmissible lung cancer caused by Jaggsiekte sheep retrovirus (JSRV). It is difficult to identify animals infected with JSRV but are clinically healthy. The virus does not induce a specific antibody response and, although proviral DNA sequences of JSRV can be found in mononuclear blood cells, the detection is inconsistent. The aim of this study was to investigate the presence of JSRV in the bone marrow of infected sheep and develop a more consistent screening method. Immunohistochemical examination of bone marrow samples from 8 asymptomatic JSRV-infected sheep revealed the presence of positively labelled cells. However, JSRV could not be detected by a highly sensitive polymerase chain reaction (PCR) in bone marrow aspirates periodically collected from these animals. Results suggest that JSRV-infected cells may be present in the bone marrow of symptomless animals, but the number is below the detectable level for PCR. Therefore, this technique does not seem to be helpful for preclinical diagnosis of OPA.     

Performance of outdoor seawater treatment systems for recirculation in an intensive turbot (Scophthalmus maximus) farm

Water treatment systems are mandatory in recirculating aquaculture facilities facing existing regulations, but data on system efficiency, especially for marine species, are scarce. The present work aimed at contributing to the evaluation of the effluent characteristics and the performance of a combined outdoor biological and non-biological treatment system in an intensive turbot (Scophthalmus maximus) farm, operating under different hydraulic regimes. A preliminary study on the biofilter bacterial populations was also undertaken. Changes in effluent characteristics with pumping, season of the year and fish biomass were observed. The treatment system showed performance instability under the conditions assayed (outdoors, changeable recycle rates). Maximum removal of solids was observed in winter, with microscreen or biological filtration (up to 60%) and nitrite removal (40–98%) was achieved with ozonation. Reduction in ammonium levels was higher in summer, either mechanically (74%) or biologically (33%). Phosphate removal was higher in winter with both systems (37 and 60%, respectively). Compliance with Portuguese discharge standards was achieved. For improvements in the treatment loop, further studies on biofilter bacteria under outdoor conditions are needed, and biological denitrification is encouraged. This revised version was published online in August 2006 with corrections to the Cover Date.     

The effect of dietary vitamin K (phylloquinone and menadione) levels on the vertebral formation in mummichog Fundulus heteroclitus

SUMMARY: The effect of different dosages of vitamin K, either as phylloquinone or menadione sodium bisulfite (MSB), on vertebrae formation during growth of mummichog Fundulus heteroclitus was investigated. Diets without vitamin K caused a significantly higher incidence of bone deformity in fish than diets supplemented with MSB or phylloquinone ( P < 0.01). These results indicate that vitamin K is essential for mummichog. A massive dose of MSB caused a high incidence of bone deformity compared with a lower dose, while an increased dose of phylloquinone brought about no significant difference in deformity incidence. The vertebral deformity was significantly higher ( P < 0.01) in fish fed a vitamin K-free diet (Control Group) and MSB-supplemented diet (Group MSB25) compared to fish fed a phylloquinone-supplemented diet (Group PK1). These results indicate that phylloquinone is more suitable than MSB as a vitamin K source in fish feed.     

Reproductive cycle of yellow sea bream <Emphasis Type="Italic">Dentex tumifrons</Emphasis> in Wakasa Bay,the Sea of Japan off central Honshu

ABSTRACT:   To determine the spawning season and reproductive cycle of the yellow sea bream, Dentex tumifrons , in Wakasa Bay, in the Sea of Japan off central Honshu, seasonal change in the gonadosomatic index ( GSI ) was examined and histological observations of both ovaries and testes were conducted. The female GSI gradually increased from May, maintained a high level between August and October and then rapidly decreased. The histological observations of the ovary showed that vitellogenesis began from May and developed until September. Spent fish began to be observed in October and all the fish collected in December were spent. There was only one peak in the seasonal change in GSI of males and females in Wakasa Bay. Thus, yellow sea bream spawn only in autumn in Wakasa Bay. Differences in the reproductive cycle and number of spawning seasons were found between the two different latitudinal areas, Wakasa Bay and the East China Sea (low latitudinal area) where the spawning seasons were spring and autumn. An increase in water temperature is an important factor to initiate gonadal maturation (recrudescence) and is terminated by the short day-length in Wakasa Bay.     

Patterns of effects of arbuscular mycorrhizal fungi on plants grown in contaminated soil

Arbuscular mycorrhizal fungi (AMF) are integral functioning parts of plant root systems and are widely recognized for enhancing plant growth on severely disturbed sites, including those contaminated with heavy metals. However, the generality of detailed patterns observed for their influence on various metals and oxidative‐stress parameters in multiple plant species is not clarified. The goal of this study was to investigate the patterns of metal‐stress alleviation by AMF in four plant species. For this purpose, clover, sunflower, mustard, and phacelia were inoculated with Glomus intraradices and compared to noninoculated plants grown under heavy metal–stressed conditions. The study focused on the effect of AMF inoculation on plant biomass, assimilating pigments, total protein, superoxide dismutase and peroxidase activity, lipid peroxidation and As, Cd, Co, Cu, Fe, Mn, P, Pb, U, and Zn contents. As a result of inoculation very different patterns of variation were obtained for concentrations of elements and for biochemical parameters in plants. The particular effect of AMF inoculation on plants was species‐ and metal‐specific, although there was a general enhancement of plant growth.     

Nanopore sequencing for the detection and identification of Xylella fastidiosa subspecies and sequence types from naturally infected plant material

Xylella fastidiosa (Xf) is a gram-negative bacterial plant pathogen that can infect over 500 plant species. While it is endemic in America, X. fastidiosa subsp. pauca was reported for the first time in Europe in 2013 on olive trees in southern Italy. The availability of fast, sensitive, and reliable diagnostic tools is indispensable for managing current and future outbreaks of Xf. In this paper, we use the OXford Nanopore Technologies (ONT) MinION platform for detecting and identifying Xf at species, subspecies, and sequence type (ST) level. Two workflows were developed: the first one provided a “shotgun” strategy, that is, exploring the possibility of detecting Xf within DNA extracted from plant samples. This allowed detection of Xf by direct DNA sequencing and identifying the subspecies only in samples with high bacterial levels. Nanopore amplicon sequencing was pursued as a second workflow. This consists of PCR amplification of a set of seven multilocus sequence typing (MLST) fragments, officially adopted for identifying Xf at type strain level, followed by Nanopore-sequencing of the amplicons and an ad hoc pipeline to generate MLST consensus calls. This combined approach, which takes only a few hours, allowed the detection and identification of Xf at ST level in plant material with low bacterial infection.     

Morphological and immunohistochemical features of Cryptosporidium andersoni in cattle

Light and electron microscopic features and immunohistochemical features of Cryptosporidium andersoni (C. andersoni) and host reaction in the mucosa were studied. Although the affected cattle demonstrated no apparent clinical signs, a severe infection of C. andersoni was observed in the abomasum. C. andersoni were round in shape, measured 6-8 microm in size and were mainly observed to be freely located in the gastric pits, being attached in occasional cases to the surface of the abomasum epithelium. Frequent inflammatory cells had infiltrated the lamina propria of the affected mucosa, and frequent mitotic figures were observed in epithelial cells at the dilated isthmus. To access the cell kinetics, the number of epithelial cells infected with C. andersoni were counted and compared with noninfected cattle. The number of gastric pit cells in infected cattle was significantly higher than that in the controls. The number of proliferative cells determined by the Ki-67 antigen in C. andersoni infected cattle was also significantly higher than that in the controls. Transmission electron microscopy and scanning electron microscopy revealed that the morphology of the C. andersoni organism was common to those of other Cryptosporidium spp. Immunohistochemically, several commercial antibodies against Cryptosporidium spp. showed positive reactions at the wall of these oocysts or parasitophorous vacuoles. This report is possibly the first to discuss the prominent hyperplasia of the abomasum mucosa, as well as morphologic features of C. andersoni in cattle.