Development of biomarkers inAporrectodea caliginosa andLumbricus rubellus for detecting exposure to benzo[a]pyrene

Objectives and Methods. The sensitivity of two biomarkers, the neutral red retention assay (NRRA) and cytochrome P450 were evaluated in the earthwormAporrectodea caliginosa for their potential to detect exposure to benzo[a]pyrene (BaP). Cytochrome P450 was also evaluated in the earthwormLumbricus rubellus, and measured using the substrate ethoxycoumarin. Optimal assay conditions (pH, and temperature) were determined, followed by exposure of earthworms to 20 mg/kg BaP (a typical concentration at contaminated sites in New Zealand). Ethoxycoumarin-O-dealkylase (ECOD) activity was measured after 7, 14, and 21 days exposure. The NRRA was evaluated in earthworms exposed to 0.2, 20, and 100 mg BaP/kg, and biomarker responses were compared with effects on body weight. Results and Discussion Benzo[a]pyrene failed to induce ECOD activity in either earthworm species, and therefore it is not useful as a biomarker of BaP exposure and was not evaluated further. In all cases, the NRRA was significantly affected in the absence of any effects on earthworm body weight, indicating that this assay can detect exposure to BaP at a range of concentrations comparable to those found at contaminated sites. The NRRA should be linked to reproductive endpoints, then it can be used as an early warning indicator of adverse effects. Establishing biomarker stability under environmental conditions is an important step in biomarker development. Therefore, the effects of soil type (sandy soil, silt loam, and a clay soil), moisture content (15–30%), and temperature (5–20°) on the NRRA were determined. In all cases, there was no effect on the NRRA, indicating that this assay is very stable under varying environmental conditions. Conclusion and Outlook In conclusion, cytochrome P450 activity does not appear to be a useful indicator of PAH exposure in eitherA. caliginosa or L.rubellus, and due to the inherently low activity, it is not suitable as a routine biomarker for detecting environmental contamination by these compounds. In contrast, the NRRA in the earthworm A.caliginosa is a promising indicator of PAH exposure at the concentrations likely to be found in contaminated sites in New Zealand, and therefore has potential for evaluating these contaminated sites. If the NRRA can be linked to ecologically important life-cycle endpoints, such as reproduction, then it could be used as an early warning indicator of adverse effects at contaminated sites, i.e., by measuring biomarker responses in earthworms from a ‘contaminated area’ and comparing these with earthworms from a matched control area.     

Errors in repeated measurements of soil water content in pots using a ThetaProbe

Abstract. The accuracy of a ThetaProbe (Delta-T Devices Ltd, UK) to obtain repeated measures of soil water content in pot plants was tested. This alternative to balance determinations led to a large underestimation of water content, varying from 12.2 to 21.8% of the total water content, depending on soil type.     

Activity of spinosad on stored-tobacco insects and persistence on cured tobacco stripst

Every year raw tobacco and manufactured tobacco products are lost to two major storage pests, the cigarette beetle, Lasioderma serricorne (F) and the tobacco moth, Ephestia elutella (Hiibner). Post-harvest management of both insects is achieved through sanitation, insect monitoring and fumigation with phosphine. However, insect resistance to phosphine and control failures have been reported, and fumigants are under constant regulatory pressure. Here we report the evaluation of spinosad, a bioinsecticide derived from the fermentation of the soil micro-organism Saccharopolyspora spinosa Mertz & Yao. Spinosad was first registered in 1997 and is now widely used as a field pest control agent on many crops, including tobacco. The insecticidal activity of the fermentation product (technical spinosad, TS) was measured by diet incorporation assays against L serricorne and E elutella larvae. Mortality levels were determined on newly hatched larvae and over the whole insect life cycle. For both species, no emergence of adult insects was observed in cured tobacco sprayed with 50mg TS kg(-1) and inoculated with eggs or newly hatched larvae. These results indicated that spinosad has potential for the control of both species in stored tobacco, since 100% control of both pests could be achieved at 50 mg TS kg(-1), and with almost full control (90-95%) at 10 mg kg(-1). We also monitored the stability of the product on cured tobacco. The original concentration of the main active component of TS, spinosyn A, did not change significantly over 18 months, indicating no loss of spinosad during a typical leaf storage period of time. Bioassays against larvae confirmed that the bioinsecticidal activity of spinosad was retained.     

Changes in glucosinolate concentrations, myrosinase activity, and production of metabolites of glucosinolates in cabbage (Brassica oleracea Var. capitata) cooked for different durations

In cabbage, glucosinolates such as sinigrin are hydrolyzed by plant myrosinase to allyl isothiocyanate (AITC), allyl cyanide, and, in the presence of an epithiospecifier protein, 1-cyano-2,3-epithiopropane (CEP). Isothiocyanates have been implicated in the cancer-protective effects of Brassica vegetables. The effect of processing on the hydrolysis of glucosinolates was investigated in cabbage. Cabbage was steamed or microwaved for six time durations over 7 min. Glucosinolate concentrations were slightly reduced after microwave cooking (P < 0.001) but were not influenced after steaming (P < 0.05). Myrosinase activity was effectively lost after 2 min of microwave cooking and after 7 min of steaming. Hydrolysis of residual glucosinolates following cooking yielded predominantly CEP at short cooking durations and AITC at longer durations until myrosinase activity was lost. Lightly cooked cabbage produced the highest yield of AITC on hydrolysis in vitro, suggesting that cooking Brassica vegetables for a relatively short duration may be desirable from a health perspective.     

Primary Aspergillus and Fusarium keratitis in a Holstein cow

A 5-year-old female Holstein cow was examined for ocular discharge, periorbital swelling, an area of full-thickness corneal cellular infiltrate, fibrin, hypopyon, diffuse corneal edema, and miosis. The patient was diagnosed with a corneal stromal abscess and secondary anterior uveitis. Histopathology, mycotic culture, and polymerase chain reaction positive for Aspergillus and Fusarium DNA confirmed the presence of fungal infection. Response to therapy was adequate, and follow-up with the patient 1 year after diagnosis revealed a focal area of corneal fibrosis.     

Detection of porcine circovirus genotypes 2a and 2b in aborted foetuses from infected swine herds in the State of S?o Paulo,Brazil

Background

Porcine circovirus type 2 (PCV2) has been associated with several disease complexes, including reproductive failure. The aim of this study was to identify the subtypes of PCV2 that are associated with reproductive failure in pigs from the State of São Paulo, Brazil and to investigate co-infections with other infectious organisms.

Findings

Samples of 168 aborted foetuses or mummified foetuses from five farrow-to-finish swine farms known to be infected with PCV2 and located in the State of São Paulo were tested for PCV2 by polymerase chain reaction (PCR). Positive samples were additionally tested for porcine parvovirus (PPV), Leptospira spp. and Brucella spp. by PCR. PCV2 was detected in 18 of the samples (10.7%). PPV, Brucella spp. and Leptospira spp were found in 2, 10 and 0 cases, respectively. Eleven PCV2 strains were sequenced and determined to be either genotype 2a (n = 1) or 2b (n = 10).

Conclusions

The findings indicate that the frequency of PCV2 infections in aborted porcine foetuses from the State of São Paulo is rather low (10.7%) and that co-infection with other pathogens is common and may be involved in PCV2 associated reproductive failure. No repeatable, characteristic amino acid motifs for regions of the PCV2 capsid protein seemed to be associated with abortion in sows.     

Probable lumbar acute non-compressive nucleus pulposus extrusion in a cat with acute onset paraparesis

A spinal cord lesion localised caudal to the L6 spinal segment was diagnosed in a 2-year-old female spayed domestic longhair cat with acute onset paraparesis. Magnetic resonance imaging findings were consistent with an acute, non-compressive nucleus pulposus extrusion of the L5-L6 intervertebral disc. The cat was successfully managed with supportive care, including cage confinement.