Pig effluent-P application can increase risk of P transport: two case studies

Land application of piggery effluent (containing urine, faeces, water, and wasted feed) is under close scrutiny as a potential source of water resource contamination with phosphorus (P). This paper investigates two case studies of the impact of long-term piggery effluent-P application to soil. A Natrustalf (Sodosol) at P1 has received a net load of 3700 kg effluent P/ha over 19 years. The Haplustalf (Dermosol) selected (P2) has received a net load of 310 000 kg P/ha over 30 years. Total, bicarbonate extractable, and soluble P forms were determined throughout the soil profiles for paired (irrigated and unirrigated) sites at P1 and P2, as well as P sorption and desorption characteristics. Surface bicarbonate (PB, 0 - 0.05 m depth) and dilute CaCl2 extractable molybdate-reactive P (PC) have been significantly elevated by effluent irrigation (P1: PB unirrigated 23±1, irrigated 290±6; PC unirrigated 0.03±0.00, irrigated 23.9±0.2. P2: PB unirrigated 72±48, irrigated 3950±1960; PC unirrigated 0.7±0.0, irrigated 443±287 mg P/kg; mean±s.d.). Phosphorus enrichment to 1.5 m, detected as PB, was observed at P2. Elevated concentrations of CaCl2 extractable organic P forms (POC; estimated by non-molybdate reactive P in centrifuged supernatants) were observed from the soil surface of P1 to a depth of 0.4 m. Despite the extent of effluent application at both of these sites, only P1 displayed evidence of significant accumulation of POC. The increase in surface soil total P (0 - 0.05 m depth) due to effluent irrigation was much greater than laboratory P sorption (>25 times for P1; >57 times for P2) for a comparable range of final solution concentrations (desorption extracts ranged from 1-5 mg P/L for P1 and 50-80 mg P/L for P2). Precipitation of sparingly soluble P phases was evidenced in the soils of the P2 effluent application area.

Evaluation of the immunogenicity of the P97R1 adhesin of Mycoplasma hyopneumoniae as a mucosal vaccine in mice.

The immunogenicity of P97 adhesin repeat region R1 (P97R1) of Mycoplasma hyopneumoniae, an important pathogenesis-associated region of P97, was evaluated in mice as a mucosal vaccine. Mice were immunized orally with attenuated Salmonella typhimurium aroA strain CS332 harbouring a eukaryotic or prokaryotic expression vector encoding IP97R1. Local and systemic immune responses were analysed by ELISA on mouse sera, lung washes and splenocyte supernatants following splenocyte stimulation with specific antigens in vitro. Although no P97R1-specific antibody responses were detected in serum and lung washes, significant gamma interferon was produced by P97R1-stimulated splenocytes from mice immunized orally with S. typhimurium aroA harbouring either expression system, indicating induction of a cell-mediated immune response. These results suggested that live bacterial vectors carrying DNA vaccines or expressing heterologous antigens preferentially induce a Th1 response. Surprisingly, however, mice immunized with the vaccine carrier S. typhimurium aroA CS332 induced serum IgG, but not mucosal IgA, against P97R1 or S. typhimurium aroA CS332 whole-cell lysate, emphasizing the importance of assessing the suitability of attenuated S. typhimurium antigen-carrier delivery vectors in the mouse model prior to their evaluation as potential vaccines in the target species, which in this instance was pigs.

Screening bacterial metabolites for inhibitory effects against Batrachochytrium dendrobatidis using a spectrophotometric assay

Certain bacteria present on frog skin can prevent infection by the pathogenic fungus Batrachochytrium dendrobatidis (Bd), conferring disease resistance. Previous studies have used agar-based in vitro challenge assays to screen bacteria for Bd-inhibitory activity and to identify candidates for bacterial supplementation trials. However, agar-based assays can be difficult to set up and to replicate reliably. To overcome these difficulties, we developed a semi-quantitative spectrophotometric challenge assay technique. Cell-free supernatants were prepared from filtered bacterial cultures and added to 96-well plates in replicated wells containing Bd zoospores suspended in tryptone-gelatin hydrolysate-lactose (TGhL) broth medium. Plates were then read daily on a spectrophotometer until positive controls reached maximum growth in order to determine growth curves for Bd. We tested the technique by screening skin bacteria from the Australian green-eyed tree frog Litoria serrata. Of bacteria tested, 31% showed some degree of Bd inhibition, while some may have promoted Bd growth, a previously unknown effect. Our cell-free supernatant challenge assay technique is an effective in vitro method for screening bacterial isolates for strong Bd-inhibitory activity. It contributes to the expanding field of bioaugmentation research, which could play a significant role in mitigating the effects of chytridiomycosis on amphibians around the world.