Npro of Bungowannah virus exhibits the same antagonistic function in the IFN induction pathway than that of other classical pestiviruses

Bungowannah virus is the most divergent atypical pestivirus that had been detected up to now, and does not fit into any of the four approved species: Bovine viral diarrhea virus type 1 (BVDV-1) and type 2 (BVDV-2), Classical swine fever virus (CSFV) and Border disease virus (BDV). However, the presence of N^pro and E^rns coding regions, which are unique to pestiviruses, provides clear evidence of a pestivirus. Nevertheless, the amino acid identity of Bungowannah virus N^pro and BVDV-1 N^pro (strain CP7) is only 51.5%. By using a BVDV-1 backbone, a novel chimeric construct was generated, in which the genomic region encoding the non-structural protein N^pro was replaced by that of Bungowannah virus (CP7_N^pro-Bungo). In vitro studies of CP7_N^pro-Bungo revealed autonomous replication with the same efficacy as the BVDV backbone CP7 and infectious high-titer virus could be collected. In order to compare the ability of interferon (IFN) suppression, two reporter gene assays, specific for type-I IFN, were carried out. In virus-infected cells, no significant difference in blocking of IFN expression between the parental virus CP7, Bungowannah virus and the chimeric construct CP7_N^pro-Bungo could be detected. In contrast, an N^pro deletion mutant showed an impaired replication in bovine cells and a marked type-I IFN response.Taken together, our findings reveal the compatibility of non-structural protein N^pro of atypical Bungowannah virus with a BVDV type 1 backbone and its characteristic feature as an inhibitor of type-I IFN induction with an inhibitor-activity comparable to other pestiviruses.     

Comparison of levels and duration of detection of antibodies to bovine viral diarrhea virus 1, bovine viral diarrhea virus 2, bovine respiratory syncytial virus,bovine herpesvirus 1, and bovine parainfluenza virus 3 in calves fed maternal colostrum or a colostrum-replacement product

Colostrum-replacement products are an alternative to provide passive immunity to neonatal calves; however, their ability to provide adequate levels of antibodies recognizing respiratory viruses has not been described. The objective of this study was to compare the serum levels of IgG at 2 d of age and the duration of detection of antibodies to bovine viral diarrhea virus 1 (BVDV-1), bovine viral diarrhea virus 2 (BVDV-2), bovine respiratory syncytial virus (BRSV), bovine herpesvirus 1 (BHV-1), and bovine parainfluenza virus 3 (BPIV-3) in calves fed maternal colostrum (MC) or a colostrum replacement (CR) at birth. Forty newborn male Holstein calves were assigned to the CR or the MC group. Group CR (n = 20) received 2 packets of colostrum replacement (100 g of IgG per 470-g packet), while group MC (n = 20) received 3.8 L of maternal colostrum. Blood samples for detection of IgG and virus antibodies were collected from each calf at birth, at 2 and 7 d, and monthly until the calves became seronegative. Calves in the MC group had greater IgG concentrations at 2 d of age. The apparent efficiency of absorption of IgG was greater in the MC group than in the CR group, although the difference was not significant. Calves in the CR group had greater concentrations of BVDV neutralizing antibodies during the first 4 mo of life. The levels of antibodies to BRSV, BHV-1, and BPIV-3 were similar in the 2 groups. The mean time to seronegativity was similar for each virus in the 2 groups; however, greater variation was observed in the antibody levels and in the duration of detection of immunity in the MC group than in the CR group. Thus, the CR product provided calves with more uniform levels and duration of antibodies to common bovine respiratory viruses.     

Prion protein genotypes of sheep as determined from 3343 samples submitted from Ontario and other provinces of Canada from 2005 to 2012

This study analyzed sheep prion protein (PrP) genotypes of samples submitted from Ontario and other provinces of Canada to the Animal Health Laboratory at the University of Guelph, Guelph, Ontario, between 2005 and 2012. In Ontario, the proportion of scrapie-resistant sheep increased from 2005 to 2012 as evidenced by an increase in the ARR haplotype. When Canadian provinces (Alberta, Ontario, Quebec, and Nova Scotia) were compared from 2008 to 2012, a high proportion of scrapie-resistant sheep was found in all the provinces. The proportions of resistant sheep were lower in Alberta and Quebec than in Ontario and Nova Scotia. Alberta had higher proportions of susceptible sheep and a higher frequency of VRQ alleles, and Quebec had a higher frequency of the ARQ allele.     

Pathology and Neurotoxicity in Dogs after Repeat Dose Exposure to a Serotonin 5-HT1B Inhibitor

AZD3783, a cationic amphiphilic drug and a potent inhibitor of the 5-hydroxytryptamine (5-HT_1B) receptor, was explored as a potential treatment for depression. To support clinical trials, repeat dose toxicity studies in rats and dogs were conducted. Here we report toxicity findings in dogs after dosing from 1 to 3 months. In the 1-month study, there were minimal neuronal vacuolation in the brain, a marked increase in liver enzymes accompanied by hepatocellular degeneration/necrosis and phospholipidosis (PLD), and PLD/cholecystitis in the gallbladder of animals dosed at 47 mg/kg/day. In the 3-month study, neurotoxicity resulted in euthanasia of one animal dosed at 30 mg/kg/day after 86 days. Extensive pathologic changes were seen in all animals in retina epithelium (inclusion bodies), brain (neuronal vacuolation, degeneration, or necrosis and nerve fiber degeneration), spinal ganglia (vacuolation, degeneration, or necrosis), as well as sciatic and optic nerves (degeneration). Pigment-laden macrophages were observed in the lung, kidney, liver, gallbladder, bone marrow, gastrointestinal tract, and lymphoid tissues. Also seen were vitrel and retinal hemorrhage in the eyes. A brain concentration and pathology study showed that the concentration of AZD3783 in the brain was approximately 4 times higher than in the plasma after 4 weeks of dosing, however, they were similar in all regions examined, and did not correlate with areas with pathologic findings. Our findings with AZD3783 in dogs have not been reported previously with other CNS compounds that effect through serotonergic pharmacology.     

Seasonal changes in proteolytic enzymes of yellowtail Seriola quinqueradiata (Temminck & Schlegel; Carangidae) fed extruded diets containing different protein and energy levels

Seasonal variations in pepsin, trypsin and chymotrypsin activities and protein digestibility were studied in yellowtails (Seriola quinqueradiata) reared for 1 year with extruded diets containing different levels of protein. Trypsin and chymotrypsin storage levels in the digestive tissues of starved fish were affected by seasonal changes in water temperature. Actual digestion activities of trypsin and chymotrypsin were low at lower water temperatures, but pepsin activity in the stomach tissue was not affected by low temperatures. On the other hand, trypsin and chymotrypsin activities in the intestinal contents were higher during lower water temperature months, while pepsin activity in the stomach contents was low at lower water temperatures. Apparent protein digestibility (APD) did not differ among the dietary treatments in the higher water temperature months, while in colder months it was higher in fish fed diet 1 than in fish fed diets 2 and 3. The APD values reflected pepsin activity in the stomach contents in all sampling months. Therefore, lower APD in colder months seems to be attributed to lower protease activity in the gastric digesta, implying that enhanced pepsin secretion from the stomach tissue might improve protein digestibility and growth performance in yellowtails during winter.     

Ammonia distribution and excretion in fish

This paper reviews the literature concerning ammonia production, storage and excretion in fish. Ammonia is the end product of protein catabolism and is stored in the body of fish in high concentrations relative to basal excretion rates. Ammonia, if allowed to accumulate, is toxic and is converted to less toxic compounds or excreted. Like other weak acids and bases, ammonia is distributed between tissue compartments in relation to transmembrane pH gradients. NH₃ is generally equilibrated between compartments but NH₄ ⁺ is distributed according to pH. Ammonia is eliminated from the blood upon passage through the gills. The mechanisms of branchial ammonia excretion vary between different species of fish and different environments, and primarily involves NH₃ passive diffusion and NH₄ ⁺/Na⁺ exchange. Water chemistry near the gill surface may also be important to ammonia excretion, but a more accurate measurement of the NH₃ gradient across the gill epithelium is required before a more detailed analysis of NH₃ and NH₄ ⁺ excretion can be made.     

Manipulations of Shrimp Embryos: A Viable Technique for the Removal of the Hatching Envelope

The hatching envelope elevated around the spawned eggs of penaeoidean eggs presents a formidable barrier for manipulation of the embryo. By spawning eggs into seawater containing 3-amino-1,2,4-triazole and subsequently passing the eggs through Nitex screening approximately 45 min after spawning, the hatching envelope can be effectively removed with little or no damage to the embryos. The time and extent to which the envelope is removed can be varied. Embryos treated in this manner continue to develop and the method provides a new technique with which investigators can gain access to the surface of the egg or embryo.     

Changes in the Solubility of Corn Proteins through Interaction with the Arabinoxylans in Extruded Nixtamalized Corn Flour Treated with Xylanase

The extrusion process allows the production of nixtamalized corn flour rich in arabinoxylans, which help to prevent cardiovascular and intestinal diseases. During extrusion, physiochemical properties of nixtamalized corn flour are negatively modified. The use of enzymes such as xylanase in order to obtain nixtamalized corn flour using extrusion has been studied as an alternative to reduce these changes in corn flour tortilla. The aim of this research was to evaluate changes in protein solubility of extruded nixtamalized corn flour with and without different concentrations of xylanase enzyme (0.05, 0.075, and 0.1 %, w/w). Soluble proteins of each corn flour were extracted and analyzed by SE-HPLC, while insoluble proteins were determined by the combustion method. In addition, each corn flour was analyzed by scanning electron microscopy (SEM). Results showed that the extruded nixtamalized corn flour, with and without xylanase, increased the protein solubility, and this effect was lower in extruded nixtamalized corn flour with xylanase. Insoluble protein diminished in corn flours either with or without xylanase enzyme. The addition of xylanase reduces the effect that the extrusion process has on the solubility proteins of extruded nixtamalized corn flour.