On-farm Campylobacter and Escherichia coli in commercial broiler chickens: Re-used bedding does not influence Campylobacter emergence and levels across sequential farming cycles

Limitations in quality bedding material have resulted in the growing need to re-use litter during broiler farming in some countries, which can be of concern from a food-safety perspective. The aim of this study was to compare the Campylobacter levels in ceca and litter across three litter treatments under commercial farming conditions. The litter treatments were (a) the use of new litter after each farming cycle; (b) an Australian partial litter re-use practice; and (c) a full litter re-use practice. The study was carried out on two farms over two years (Farm 1, from 2009–2010 and Farm 2, from 2010–2011), across three sheds (35,000 to 40,000 chickens/shed) on each farm, adopting three different litter treatments across six commercial cycles. A random sampling design was adopted to test litter and ceca for Campylobacter and Escherichia coli, prior to commercial first thin-out and final pick-up. Campylobacter levels varied little across litter practices and farming cycles on each farm and were in the range of log 8.0–9.0 CFU/g in ceca and log 4.0–6.0 MPN/g for litter. Similarly the E. coli in ceca were ∼log 7.0 CFU/g. At first thin-out and final pick-up, the statistical analysis for both litter and ceca showed that the three-way interaction (treatments by farms by times) was highly significant (P < 0.01), indicating that the patterns of Campylobacter emergence/presence across time vary between the farms, cycles and pickups. The emergence and levels of both organisms were not influenced by litter treatments across the six farming cycles on both farms. Either C. jejuni or C. coli could be the dominant species across litter and ceca, and this phenomenon could not be attributed to specific litter treatments. Irrespective of the litter treatments in place, cycle 2 on Farm 2 remained campylobacter-free. These outcomes suggest that litter treatments did not directly influence the time of emergence and levels of Campylobacter and E. coli during commercial farming.

Getting value out of your sensory testing - the difference from control test

Non-parametric difference tests such as triangle and duo-trio tests traditionally are used to establish differences or similarities between products. However they only supply the researcher with partial answers and often further testing is required to establish the nature, size and direction of differences. This paper looks at the advantages of the difference from control (DFC) test (also known as degree of difference test) and discusses appropriate applications of the test. The scope and principle of the test, panel composition and analysis of results are presented with the aid of suitable examples. Two of the major uses of the DFC test are in quality control and shelf-life testing. The role DFC takes in these areas and the use of other tests to complement the testing is discussed. Controls or standards are important in both these areas and the use of standard products, mental and written standards and blind controls are highlighted. The DFC test has applications in products where the duo-trio and triangle tests cannot be used because of the normal heterogeneity of the product. While the DFC test is a simple difference test it can be structured to give the researcher more valuable data and scope to make informed decisions about their product.

A simplified PCR test for early detection of dwarf off-types in micropropagated Cavendish bananas (Musa spp. AAA)

The dwarf somaclonal variant is a major problem affecting micropropagation of the banana cultivar Williams (Musa spp. AAA; subgroup Cavendish). This problem arises from genetic changes that occur during the tissue culture process. Early identification of this problem is difficult and propagators must wait until plants are ex vitro in order to visualise the dwarfism phenotype. In this study, we have improved a SCAR-based molecular diagnostic technique, developed by Damasco et al. [Acta Hortic. 461 (1997) 157], for the early identification of dwarf off-types. We have included a positive internal control in a multiplex PCR and adapted the technique for use with small amounts of fresh in vitro leaf material as PCR template. The control product is a 500 bp fragment from 18S rRNA and is amplified in all tissues irrespective of phenotype. The use of small in vitro leaf material removing the need for genomic DNA extraction.

Comparison of the indirect haemagglutination and gel diffusion test for serotyping Haemophilus parasuis

The aim of this study was to compare the use of indirect haemagglutination (IHA) and gel diffusion (GD) tests for serotyping Haemophilus parasuis by the Kielstein-Rapp-Gabrielson scheme. All 15 serovar reference strains, 72 Australian field isolates, nine Chinese field isolates, and seven isolates from seven experimentally infected pigs were evaluated with both tests. With the IHA test, 14 of the 15 reference strains were correctly serotyped – with serovar 10 failing to give a titre with serovar 10 antiserum. In the GD test, 13 reference strains were correctly serotyped – with antigen from serovars 7 and 8 failing to react with any antiserum. The IHA methodology serotyped a total of 45 of 81 field isolates while the GD methodology serotyped a total of 48 isolates. For 29 isolates, the GD and IHA methods gave discordant results. It was concluded that the IHA is a good additional test for the serotyping of H. parasuis by the KRG scheme if the GD methodology fails to provide a result or shows unusual cross-reactions.

A universal test to determine the integrity of RNA, and its suitability for reverse transcription, in animal tissue laboratory specimens

Degradation of RNA in diagnostic specimens can cause false-negative test results and potential misdiagnosis when tests rely on the detection of specific RNA sequence. Current molecular methods of checking RNA integrity tend to be host species or group specific, necessitating libraries of primers and reaction conditions. The objective here was to develop a universal (multi-species) quality assurance tool for determining the integrity of RNA in animal tissues submitted to a laboratory for analyses. Ribosomal RNA (16S rRNA) transcribed from the mitochondrial 16S rDNA was used as template material for reverse transcription to cDNA and was amplified using polymerase chain reaction (PCR). As mitochondrial DNA has a high level of conservation, the primers used were shown to reverse transcribe and amplify RNA from every animal species tested. Deliberate degradation of rRNA template through temperature abuse of samples resulted in no reverse transcription and amplification. Samples spiked with viruses showed that single-stranded viral RNA and rRNA in the same sample degraded at similar rates, hence reverse transcription and PCR amplification of 16S rRNA could be used as a test of sample integrity and suitability for analysis that required the sample's RNA, including viral RNA. This test will be an invaluable quality assurance tool for determination of RNA integrity from tissue samples, thus avoiding erroneous test results that might occur if degraded target RNA is used unknowingly as template material for reverse transcription and subsequent PCR amplification.

Effect of soil burial depth and wetting on mortality of diapausing larvae and patterns of post-diapause adult emergence of sorghum midge, Stenodiplosis sorghicola (Coquillett) (Diptera: Cecidomyiidae)

In south-eastern Queensland, Australia, sorghum planted in early spring usually escapes sorghum midge, Stenodiplosis sorghicola, attack. Experiments were conducted to better understand the role of winter diapause in the population dynamics of this pest. Emergence patterns of adult midge from diapausing larvae on the soil surface and at various depths were investigated during spring to autumn of 1987/88–1989/90. From 1987/88 to 1989/90, 89%, 65% and 98% of adult emergence, respectively, occurred during November and December. Adult emergence from larvae diapausing on the soil surface was severely reduced due to high mortality attributed to surface soil temperatures in excess of 40°C, with much of this mortality occurring between mid-September and mid-October. Emergence of adults from the soil surface was considerably delayed in the 1988/89 season compared with larvae buried at 5 or 10 cm which had similar emergence patterns for all three seasons. In 1989/90, when a 1-cm-deep treatment was included, there was a 392% increase in adult emergence from this treatment compared with deeper treatments. Some diapausing larvae on the surface did not emerge at the end of summer in only 1 year (1989/90), when 28.0% of the larvae on the surface remained in diapause, whereas only 0.8% of the buried larvae remained in diapause. We conclude that the pattern of emergence explains why spring plantings of sorghum in south-eastern Queensland usually escape sorghum midge attack.

The freezing characteristics of wheat at ear emergence

Wheat is occasionally exposed to freezing temperatures during ear emergence and can suffer severe frost damage. Few studies have attempted to understand the characteristics of freezing and frost damage to wheat during late development stages. It was clearly shown that wheat appears to have an inherent frost resistance to temperatures down to −5 °C but is extensively damaged below this temperature. Acclimation, whilst increasing the frost resistance of winter wheat in a vegetative state was incapable of increasing frost resistance of plants at ear emergence. It is proposed that the ability to upregulate frost resistance is lost once vernalisation requirement is fulfilled. Culms and ears of wheat were able to escape frost damage at temperatures below −5 °C by supercooling even to as low as −15 °C and evidence collected by infrared thermography suggested that individual culms on a plant froze as independent units during freezing with little or no cross ice-nucleation strategies to protect wheat from frost damage in the field appear to revolve around avoiding ice nucleation.

The vaccination-challenge trial: the gold standard test to evaluate the protective efficacy of infectious coryza vaccines

Infectious coryza is an upper respiratory tract disease of chickens with the major impact occurring in multi-age flocks. We investigated the relationship between the level of antibodies, as detected by a haemagglutination-inhibition (HI) assay, in infectious coryza-vaccinated chickens and the protection against challenge in those chickens. In one experiment, chickens given a single dose of either of two infectious coryza vaccines lacked a detectable HI response to vaccination but showed significant levels of protection 11 weeks after vaccination. In contrast, in chickens given two doses of an infectious coryza vaccine and challenged 3 weeks after the second vaccine dose, there was a strong serological response with 36/40 birds having a HI titre of 1/20 or greater. In this trial there was an apparent relationship between titre and subsequent protection, with none of the 32 chickens with a titre of 1/40 or 1/80 showing any clinical signs and only one of the same group yielding the challenge organism on culture. In contrast, three of the four vaccinated chickens with a HI titre less than 1/5 developed the typical clinical signs of coryza and yielded the challenge organism on culture. Overall, our results suggest that HI titres cannot be regarded as a definitive predictor of vaccine efficacy. We suggest that the vaccination-challenge trial is the gold standard for the evaluation of the immune response to infectious coryza vaccines.

Test of the enemy release hypothesis: the native magpie moth prefers a native fireweed (Senecio pinnatifolius) to its introduced congener (S. madagascariensis).

The enemy release hypothesis predicts that native herbivores will either prefer or cause more damage to native than introduced plant species. We tested this using preference and performance experiments in the laboratory and surveys of leaf damage caused by the magpie moth Nyctemera amica on a co-occuring native and introduced species of fireweed (Senecio) in eastern Australia. In the laboratory, ovipositing females and feeding larvae preferred the native S. pinnatifolius over the introduced S. madagascariensis. Larvae performed equally well on foliage of S. pinnatifolius and S. madagascariensis: pupal weights did not differ between insects reared on the two species, but growth rates were significantly faster on S. pinnatifolius. In the field, foliage damage was significantly greater on native S. pinnatifolius than introduced S. madagascariensis. These results support the enemy release hypothesis, and suggest that the failure of native consumers to switch to introduced species contributes to their invasive success. Both plant species experienced reduced, rather than increased, levels of herbivory when growing in mixed populations, as opposed to pure stands in the field; thus, there was no evidence that apparent competition occurred.

Lantana (Lantana camara) Seed Bank Dynamics: Seedling Emergence and Seed Survival.

Seed persistence is poorly quantified for invasive plants of subtropical and tropical environments and Lantana camara, one of the world's worst weeds, is no exception. We investigated germination, seedling emergence, and seed survival of two lantana biotypes (Pink and pink-edged red [PER]) in southeastern Queensland, Australia. Controlled experiments were undertaken in 2002 and repeated in 2004, with treatments comprising two differing environmental regimes (irrigated and natural rainfall) and sowing depths (0 and 2 cm). Seed survival and seedling emergence were significantly affected by all factors (time, biotype, environment, sowing depth, and cohort) (P < 0.001). Seed dormancy varied with treatment (environment, sowing depth, biotype, and cohort) (P < 0.001), but declined rapidly after 6 mo. Significant differential responses by the two biotypes to sowing depth and environment were detected for both seed survival and seedling emergence (P < 0.001). Seed mass was consistently lower in the PER biotype at the population level (P < 0.001), but this variation did not adequately explain the differential responses. Moreover, under natural rainfall the magnitude of the biotype effect was unlikely to result in ecologically significant differences. Seed survival after 36 mo under natural rainfall ranged from 6.8 to 21.3%. Best fit regression analysis of the decline in seed survival over time yielded a five-parameter exponential decay model with a lower asymptote approaching −0.38 (% seed survival = [( 55 − (−0.38)) • e (k • t)] + −0.38; R2 = 88.5%; 9 df). Environmental conditions and burial affected the slope parameter or k value significantly (P < 0.01). Seed survival projections from the model were greatest for buried seeds under natural rainfall (11 yr) and least under irrigation (3 yr). Experimental data and model projections suggest that lantana has a persistent seed bank and this should be considered in management programs, particularly those aimed at eradication.

Germination, emergence, and persistence of Sonchus oleraceus, a major crop weed in subtropical Australia.

Sonchus oleraceus (common sowthistle) is a dominant weed and has increased in prevalence in conservation cropping systems of the subtropical grain region of Australia. Four experiments were undertaken to define the environmental factors that favor its germination, emergence, and seed persistence. Seeds were germinated at constant temperatures between 5 and 35C and water potentials between 0 and -1.4 MPa. The maximum germination rate of 86-100% occurred at 0 and -0.2 MPa, irrespective of the temperature when exposed to light (12 h photoperiod light/dark), but the germination rate was reduced by 72% without light. At water potentials of -0.6 to -0.8 MPa, the germination rate was reduced substantially by higher temperatures; no seed germinated at a water potential >-1.0 MPa. Emergence and seed persistence were measured over 30 months following seed burial at 0 (surface), 1, 2, 5, and 10 cm depths in large pots that were buried in a south-eastern Queensland field. Seedlings emerged readily from the surface and 1 cm depth, with no emergence from below the 2 cm depth. The seedlings emerged during any season following rain but, predominantly, within 6 months of planting. Seed persistence was short-term on the soil surface, with 2% of seeds remaining after 6 months, but it increased with the burial depth, with 12% remaining after 30 months at 10 cm. Thus, a minimal seed burial depth with reduced tillage and increased surface soil water with stubble retention has favored the proliferation of this weed in any season in a subtropical environment. However, diligent management without seed replenishment will greatly reduce this weed problem within a short period.

Emergence and seed persistence of Echinochloa colona, Urochloa panicoides and Hibiscus trionum in the sub-tropical environment of north-eastern Australia.

Better understanding of seed-bank dynamics of Echinochloa colona, Urochloa panicoides and Hibiscus trionum, major crop weeds in sub-tropical Australia, was needed to improve weed control. Emergence patterns and seed persistence were investigated, with viable seeds sown at different depths in large in-ground pots. Seedlings of all species emerged between October and March when mean soil temperatures were 21-23C. However, E. colona emerged as a series of flushes predominantly in the first year, with most seedlings emerging from 0-2 cm. Urochloa panicoides emerged mostly as a single large flush in the first two years, with most seedlings emerging from 5 cm. Hibiscus trionum emerged as a series of flushes over three seasons, initially with majority from 5 cm and then 0-2 cm in the later seasons. Longevity of the grass seed was short, with <5% remaining after burial at 0-2 cm for 24 months. In contrast, 38% of H. trionum seeds remained viable after the same period. Persistence of all species increased significantly with burial depth. These data highlight that management strategies need to be tailored for each species, particularly relating to the need for monitoring, application times for control tactics, impact of tillage, and time needed to reduce the seed-bank to low numbers.

Test and develop sustainable and profitable grazing strategies to manage for rainfall variability

The Wambiana grazing trial started in 1997 to test and develop sustainable and profitable grazing strategies to manage for rainfall variability. It is important that this trial continue as the results are still relatively short-term relative to rainfall cycles and significant treatment changes are still occurring. This new proposal will maintain baseline treatments but will modify others based on trial learning’s to date. It builds on treatment differences and evidence collected over the last 12 years to deliver evidence-based guidelines and principles for sustainable and productive management. The trial also links to other projects modelling water quality, climate change, methane emissions and soil C sequestration on grazing lands.

Development of commercial application of an avocado fruit robustness test

Fruit quality is one of the major factors limiting growth in avocado retail sales. Avocado growers are often unaware of their end-use fruit quality since quality problems only manifest upon fruit ripening and growers receive limited feedback from the supply chain. If growers were aware of their expected fruit quality they would be equipped to make better marketing decisions and if necessary to take remedial actions to improve their fruit quality. Avotest is being developed as a quick and easy method of determining expected end-use fruit quality before the start of the commercial fruit harvest. The test aims at distinguishing between blocks with robust fruit and those with less robust fruit. The test could also be used to predict the resulting fruit quality after the implementation of new farming practices.

Post head-emergence frost resistance of barley genotypes in the northern grain region of Australia.

Post head-emergence frost causes substantial losses for Australian barley producers. Varieties with improved resistance would have a significant positive impact on Australian cropping enterprises. Five barley genotypes previously tested for reproductive frost resistance in southern Australia were tested, post head-emergence, in the northern grain region of Australia and compared with the typical northern control cultivars, Gilbert and Kaputar. All tested genotypes suffered severe damage to whole heads and stems at plant minimum temperatures less than -8degreesC. In 2003, 2004 and 2005, frost events reaching a plant minimum temperature of ~-6.5degreesC did not result in the complete loss of grain yield. Rather, partial seed set was observed. The control genotype, Gilbert, exhibited seed set that was greater than or equal to that of any genotype in each year, as did Kaputar when tested in 2005. Thus, Gilbert and Kaputar were at least as resistant as any tested genotype. This contrasts with trial results from the southern grain region where Gilbert was reported to be less resistant than Franklin, Amagi Nijo and Haruna Nijo. Hence, rankings for post head-emergence frost damage in the northern grain region differ from those previously reported. These results indicate that Franklin, Amagi Nijo and Haruna Nijo are not likely to provide useful sources of frost resistance or markers to develop improved varieties for the northern grain region of Australia.