We report the development and isolation of a cell line, termed HepAD38, that replicates human hepatitis B virus (HBV) under conditions that can be regulated with tetracycline. In the presence of the antibiotic, this cell line is free of virus due to the repression of pregenomic (pg) RNA synthesis. Upon removal of tetracycline from the culture medium, the cells express viral pg RNA, accumulate subviral particles in the cytoplasm that contain DNA intermediates characteristic of viral replication, and secrete virus-like particles into the supernatant. Since the HepAD38 cell line can produce high levels of HBV DNA, it should be useful for analyses of the viral replication cycle that depend upon viral DNA synthesis in a synchronized fashion. In addition, this cell line has been formatted into a high-throughput, cell-based assay that permits the large-scale screening of diverse compound libraries for new classes of inhibitors of HBV replication.

The aims of this study were to: (i) determine the chemical composition of 11 samples of crude glycerol collected from seven Australian biodiesel manufacturers; and (ii) examine the effects of increasing levels of crude glycerol fed to growing-finishing pigs on performance, plasma metabolites and meat quality at slaughter. Chemical composition of crude glycerol samples varied considerably; glycerol content ranged between 38 and 96%, with some samples containing up to 29% ash and 14% methanol. One of these samples (76.1% glycerol, 1.83% methanol) was then fed to 64 female pigs (50.9 ± 5.55 kg; mean ± s.d.) allocated to one of five dietary treatments (0, 4, 8, 12 and 16% crude glycerol) until they reached 105 kg liveweight. There were no statistical differences in performance indices with increasing levels of added glycerol, although there was an unexpectedly high variation between treatments. Blood glycerol levels were unaffected by diet in week two of the experiment, but increased linearly (P < 0.001) with increasing levels of dietary glycerol before slaughter. The inclusion of crude glycerol did not influence any meat quality parameters at slaughter (P > 0.05). Diets containing added crude glycerol were less dusty after mixing, but diets that contained 8, 12 and 16% glycerol all formed a firm aggregate within 24 h of mixing that presented some feeding difficulties. This might restrict inclusion of glycerol in mash diets to dietary levels less than 8%. Furthermore, levels of residues such as methanol and ash should be monitored to prevent excessive amounts of these compounds in pig diets.

Thirty-six sows were used to study responses of milk production, milk composition, and N balance to six concentrations of dietary CP ranging from 63 to 238 g/kg (4.4 to 15.1 g of lysine/kg) during the first lactation. Sows, on average, were 149.3 kg live weight and had 26.1 mm of back fat at P1 (which is 45 mm from the midline at the level of the last rib) immediately after parturition. During lactation, all sows suckled nine pigs each and were offered up to 4,000 g of feed daily; diets contained similar balances of amino acids and similar amounts of DE (3.56 to 3.63 Mcal/kg). Nitrogen balance trials were conducted during early and late lactation and 5-d collection periods commenced on d 10 and 24 of lactation, respectively. During both periods of lactation, there were significant positive linear relationships between the level of dietary protein and milk yield and contents of fat and total solids in milk. Milk yield increased from 7.79 to 9.91 kg/d and from 7.02 to 8.90 kg/d, whereas total solids in milk increased from 199 to 225 g/kg and from 202 to 228 g/kg during early and late lactation, respectively, in response to increasing level of dietary protein from 63 to 238 g of CP/kg. A two-phase linear regression model used to describe the relationship between N balance and dietary CP level established that sows required a diet containing > or = 202 g of CP/kg or 12.8 g of lysine/kg to maximize N balances during both stages of lactation.(ABSTRACT TRUNCATED AT 250 WORDS)

Sixty pigs were used to investigate the effects of two levels of dietary ractopamine (RAC; 0 and 20 mg/kg) and three sex types (SEX; boars, gilts, and barrows) on performance over the live weight range 60 to 90 kg. Pigs were housed in individual pens and allowed ad libitum access to a diet containing 3.466 Mcal of DE and 10.7 g of lysine/kg. Control boars exhibited faster and more efficient growth and deposited more protein and less fat than gilts or barrows. The RAC increased ADG by 17 and 21% in gilts and barrows but not in boars. Feed intake was not altered by dietary RAC. Dietary RAC increased the rate of protein deposition by 15, 42, and 41% in boars, gilts, and barrows, respectively. Nevertheless, the daily rate of protein deposition was greatest in RAC-treated boars. The RAC tended to reduce the daily rate of fat deposition by 21% in boars but not in gilts or barrows. Carcass protein content increased by 5% and fat content decreased by 8% in response to RAC. These improvements in carcass composition occurred without compromising meat quality. Results show that RAC is a potent stimulator of protein deposition in finishing pigs. However, increased protein deposition is not necessarily at the expense of fat deposition.

AT-61, a member of a novel class of phenylpropenamide derivatives, was found to be a highly selective and potent inhibitor of human hepatitis B virus (HBV) replication in four different human hepatoblastoma cell lines which support the replication of HBV (i.e., HepAD38, HepAD79, 2.2.15, and transiently transfected HepG2 cells). This compound was equally effective at inhibiting both the formation of intracellular immature core particles and the release of extracellular virions, with 50% effective concentrations ranging from 0.6 to 5.7 μM. AT-61 (27 μM) was able to reduce the amount of HBV covalently closed circular DNA found in the nuclei of HepAD38 cells by >99%. AT-61 at concentrations of >27 μM had little effect on the amount of viral RNA found within the cytoplasms of induced HepAD38 cells but reduced the number of immature virions which contained pregenomic RNA by >99%. The potency of AT-61 was not affected by one of the mutations responsible for (−)-β-l-2′,3′-dideoxy-3′ thiacytidine (3TC) resistance in HBV, and AT-61 acted synergistic with 3TC to inhibit HBV replication. AT-61 (81 μM) was not cytotoxic or antiproliferative to several cell lines and had no antiviral effect on woodchuck or duck HBV, human immunodeficiency virus type 1, herpes simplex virus type 1, vesicular stomatitis virus, or Newcastle disease virus. Therefore, we concluded that the antiviral activity of AT-61 is specific for HBV replication and most likely occurs at one of the steps between the synthesis of viral RNA and the packaging of pregenomic RNA into immature core particles.