Probiotic Bacillus amyloliquefaciens Strain H57 Improves the Performance of Pregnant and Lactating Ewes Fed a Diet Based on Palm Kernel Meal

Probiotic supplements are single or mixed strain cultures of live microorganisms that benefit the host by improving the properties of the indigenous microflora (Seo et al 2010). In a pilot study at the University of Queensland, Norton et al (2008) found that Bacillus amyloliquefaciens Strain H57 (H57), primarily investigated as an inoculum to make high-quality hay, improved feed intake and nitrogen utilisation over several weeks in pregnant ewes. The purpose of the following study was to further challenge the potential of H57 -to show it survives the steam-pelleting process, and that it improves the performance of ewes fed pellets based on an agro-industrial by-product with a reputation for poor palatability, palm kernel meal (PKM), (McNeill 2013). Thirty-two first-parity White Dorper ewes (day 37 of pregnancy, mean liveweight = 47.3 kg, mean age = 15 months) were inducted into individual pens in the animal house at the University of Queensland, Gatton. They were adjusted onto PKM-based pellets (g/kg drymatter (DM): PKM, 408; sorghum, 430; chick pea hulls, 103; minerals and vitamins; Crude protein, 128; ME: 11.1MJ/kg DM) until day 89 of pregnancy and thereafter fed a predominately pelleted diet incorporating with or without H57 spores (10 9 colony forming units (cfu)/kg pellet, as fed), plus 100g/ewe/day oaten chaff, until day 7 of lactation. From day 7 to 20 of lactation the pelleted component of the diet was steadily reduced to be replaced by a 50:50 mix of lucerne: oaten chaff, fed ad libitum, plus 100g/ewe/day of ground sorghum grain with or without H57 (10 9 cfu/ewe/day). The period of adjustment in pregnancy (day 37-89) extended beyond expectations due to some evidence of mild ruminal acidosis after some initially high intakes that were followed by low intakes. During that time the diet was modified, in an attempt to improve palatability, by the addition of oaten chaff and the removal of an acidifying agent (NH4Cl) that was added initially to reduce the risk of urinary calculi. Eight ewes were removed due to inappetence, leaving 24 ewes to start the trial at day 90 of pregnancy. From day 90 of pregnancy until day 63 of lactation, liveweights of the ewes and their lambs were determined weekly and at parturition. Feed intakes of the ewes were determined weekly. Once lambing began, 1 ewe was removed as it gave birth to twin lambs (whereas the rest gave birth to a single lamb), 4 due to the loss of their lambs (2 to dystocia), and 1 due to copper toxicity. The PKM pellets were suspected to be the cause of the copper toxicity and so were removed in early lactation. Hence, the final statistical analysis using STATISTICA 8 (Repeated measures ANOVA for feed intake, One-way ANOVA for liveweight change and birth weight) was completed on 23 ewes for the pregnancy period (n = 11 fed H57; n = 12 control), and 18 ewes or lambs for the lactation period (n = 8 fed H57; n = 10 control). From day 90 of pregnancy until parturition the H57 supplemented ewes ate 17 more DM (g/day: 1041 vs 889, sed = 42.4, P = 0.04) and gained more liveweight (g/day: 193 vs 24.0, sed = 25.4, P = 0.0002), but produced lambs with a similar birthweight (kg: 4.18 vs 3.99, sed = 0.19, P = 0.54). Over the 63 days of lactation the H57 ewes ate similar amounts of DM but grew slower than the control ewes (g/day: 1.5 vs 97.0, sed = 21.7, P = 0.012). The lambs of the H57 ewes grew faster than those of the control ewes for the first 21 days of lactation (g/day: 356 vs 265, sed = 16.5, P = 0.006). These data support the findings of Norton et al (2008) and Kritas et al (2006) that certain Bacillus spp. supplements can improve the performance of pregnant and lactating ewes. In the current study we particularly highlighted the capacity of H57 to stimulate immature ewes to continue to grow maternal tissue through pregnancy, possibly through an enhanced appetite, which appeared then to stimulate a greater capacity to partition nutrients to their lambs through milk, at least for the first few weeks of lactation, a critical time for optimising lamb survival. To conclude, H57 can survive the steam pelleting process to improve feed intake and maternal liveweight gain in late pregnancy, and performance in early lactation, of first-parity ewes fed a diet based on PKM.

Recovery and succession in a multi-species tropical seagrass meadow following experimental disturbance: the role of sexual and asexual reproduction

Recolonisation and succession in a multi-species tropical seagrass meadow was examined by creating gaps (50×50 cm) in the meadow and manipulating the supply of sexual and asexual propagules. Measurements of leaf shoot density and estimates of above-ground biomass were conducted monthly to measure recovery of gaps between September 1995 and November 1997. Measurements of the seeds stored in the sediment (seed bank) and horizontal rhizome growth of colonising species were also conducted to determine their role in the recovery process. Asexual colonisation through horizontal rhizome growth from the surrounding meadow was the main mechanism for colonisation of gaps created in the meadow. The seed bank played no role in recolonisation of cleared plots. Total shoot density and above-ground biomass (all species pooled) of cleared plots recovered asexually to the level of the undisturbed controls in 10 and 7 months, respectively. There was some sexual recruitment into cleared plots where asexual colonisation was prevented but seagrass abundance (shoot density and biomass) did not reach the level of unmanipulated controls. Seagrass species did not appear to form seed banks despite some species being capable of producing long-lived seeds. The species composition of cleared plots remained different to the undisturbed controls throughout the 26-month experiment. Syringodium isoetifolium was a rapid asexual coloniser of disturbed plots and remained at higher abundances than in the control treatments for the duration of the study. S. isoetifolium had the fastest horizontal rhizome growth of species asexually colonising cleared plots (6.9 mm day−1). Halophila ovalis was the most successful sexual coloniser but was displaced by asexually colonising species. H. ovalis was the only species observed to produce fruits during the study. Small disturbances in the meadow led to long-term (>2 years) changes in community composition. This study demonstrated that succession in tropical seagrass communities was not a deterministic process. Variations in recovery observed for different tropical seagrass communities highlighted the importance of understanding life history characteristics of species within individual communities to effectively predict their response to disturbance. A reproductive strategy involving clonal growth and production of long-lived, locally dispersed seeds is suggested which may provide an evolutionary advantage to plants growing in tropical environments subject to temporally unpredictable major disturbances such as cyclones

The Performance of Chromium and Ytterbium Intraruminal Controlled Release Devices in Beef Cattle

The estimation of feed efficiency in beef cattle should be undertaken in the environment where the genetic potential of the animal is to be expressed. Consequently if cattle are to be pasture fed, there is a requirement for accurate feed intake measurements on individual animals in the field. This in turn requires the use of accurate faecal marker delivery systems.

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.

The effect of habitat and environmental history on otolith chemistry of barramundi Lates calcarifer in estuarine populations of a regulated tropical river

We examine the microchemistry of otoliths of cohorts of a fished shed population of the large catadromous fish, barramundi Lates calcarifer from the estuary of a large tropical river. Barramundi from the estuary of the large, heavily regulated Fitzroy River, north eastern Australia were analysed by making transects of 87Sr/86Sr isotope and trace metal/Ca ratios from the core to the outer edge. Firstly, we examined the Sr/Ca, Ba/Ca, Mg/Ca and Mn/Ca and 87Sr/86Sr isotope ratios in otoliths of barramundi tagged in either freshwater or estuarine habitats that were caught by the commercial fishery in the estuary. We used 87Sr/86Sr isotope ratios to identify periods of freshwater residency and assess whether trace metal/Ca ratios varied between habitats. Only Sr/Ca consistently varied between known periods of estuarine or freshwater residency. The relationships between trace metal/Ca and river flow, salinity, temperature were examined in fish tagged and recaptured in the estuary. We found weak and inconsistent patterns in relationships between these variables in the majority of fish. These results suggest that both individual movement history within the estuary and the scale of environmental monitoring were reducing our ability to detect any patterns. Finally, we examined fish in the estuary from two dominant age cohorts (4 and 7 yr old) before and after a large flood in 2003 to ascertain if the flood had enabled fish from freshwater habitats to migrate to the estuary. There was no difference in the proportion of fish in the estuary that had accessed freshwater after the flood. Instead, we found that larger individuals with each age cohort were more likely to have spent a period in freshwater. This highlights the need to maintain freshwater flows in rivers. About half the fish examined had accessed freshwater habitats before capture. Of these, all had spent at least their first two months in marine salinity waters before entering freshwater and some did not enter freshwater until four years of age. This contrasts with the results of several previous studies in other parts of the range that found that access to freshwater swamps by larval barramundi was important for enhanced population productivity and recruitment.

The assessment of TGGE for the detection of interspecific and intergeneric DNA-marker polymorphism within Solanaceae

RFLP markers are currently the most appropriate marker system for the identification of uncharacterised polymorphism at the interspecific and intergeneric level. Given the benefits of a PCR-based marker system and the availability of sequence information for many Solanaceous cDNA clones, it is now possible to target conserved fragments, for primer development, that flank sequences possessing interspecific polymorphism. The potential outcome is the development of a suite of markers that amplify widely in Solanaceae. Temperature gradient gel electrophoresis (TGGE) is a relatively inexpensive gel-based system that is suitable for the detection of most single-base changes. TGGE can be used to screen for both known and unknown polymorphisms, and has been assessed here, for the development of PCR-based markers that are useful for the detection of interspecific variation within Solanaceae. Fifteen markers are presented where differences between Lycopersicon esculentum and L. pennellii have been detected by TGGE. The markers were assessed on a wider selection of plant species and found to be potentially useful for the identification of interspecific and intergeneric polymorphism in Solanaceous plants.

Simulating the effects of saline and sodic subsoils on wheat crops growing on Vertosols

Soils with high levels of chloride and/or sodium in their subsurface layers are often referred to as having subsoil constraints (SSCs). There is growing evidence that SSCs affect wheat yields by increasing the lower limit of a crop's available soil water (CLL) and thus reducing the soil's plant-available water capacity (PAWC). This proposal was tested by simulation of 33 farmers' paddocks in south-western Queensland and north-western New South Wales. The simulated results accounted for 79% of observed variation in grain yield, with a root mean squared deviation (RMSD) of 0.50 t/ha. This result was as close as any achieved from sites without SSCs, thus providing strong support for the proposed mechanism that SSCs affect wheat yields by increasing the CLL and thus reducing the soil's PAWC. In order to reduce the need to measure CLL of every paddock or management zone, two additional approaches to simulating the effects of SSCs were tested. In the first approach the CLL of soils was predicted from the 0.3-0.5 m soil layer, which was taken as the reference CLL of a soil regardless of its level of SSCs, while the CLL values of soil layers below 0.5 m depth were calculated as a function of these soils' 0.3-0.5 m CLL values as well as of soil depth plus one of the SSC indices EC, Cl, ESP, or Na. The best estimates of subsoil CLL values were obtained when the effects of SSCs were described by an ESP-dependent function. In the second approach, depth-dependent CLL values were also derived from the CLL values of the 0.3-0.5 m soil layer. However, instead of using SSC indices to further modify CLL, the default values of the water-extraction coefficient (kl) of each depth layer were modified as a function of the SSC indices. The strength of this approach was evaluated on the basis of correlation of observed and simulated grain yields. In this approach the best estimates were obtained when the default kl values were multiplied by a Cl-determined function. The kl approach was also evaluated with respect to simulated soil moisture at anthesis and at grain maturity. Results using this approach were highly correlated with soil moisture results obtained from simulations based on the measured CLL values. This research provides strong evidence that the effects of SSCs on wheat yields are accounted for by the effects of these constraints on wheat CLL values. The study also produced two satisfactory methods for simulating the effects of SSCs on CLL and on grain yield. While Cl and ESP proved to be effective indices of SSCs, EC was not effective due to the confounding effect of the presence of gypsum in some of these soils. This study provides the tools necessary for investigating the effects of SSCs on wheat crop yields and natural resource management (NRM) issues such as runoff, recharge, and nutrient loss through simulation studies. It also facilitates investigation of suggested agronomic adaptations to SSCs.

Recovery of intravenously infused chromium EDTA and lithium sulphate in the urine of cattle and their use as markers to measure urine volume

A series of metabolism experiments investigated the recovery of continuous-, intravenously infused chromium complexed with ethylenediamine tetra-acetic acid (CrEDTA) and lithium sulphate in the urine of cattle with a view to using the markers to estimate urine and metabolite output in grazing cattle. The recovery of Cr in urine from these infusions was similar (90%) in metabolism trials when cattle consumed three very contrasting diets: high-grain formulated pellet, lucerne hay (Medicago sativa) or low-quality native grass hay (predominantly Heteropogon contortus). By contrast, Li recovery in urine averaged 46.3 +/- 0.40% and 72.6 +/- 0.43% for native pasture and lucerne hays, respectively, but was not constant across days. There was negligible transfer of Cr from CrEDTA in blood serum to the rumen or faeces, whereas appreciable quantities of infused Li were found in both. The ratio of urine volume estimated by spot samples and marker dilution of Cr, to urine volume measured gravimetrically, was 1.05. In grazing studies using rumen-fistulated (RF) steers grazing seven different tropical and temperate grass and legume pastures, the ratio of concentrations of purine derivatives (PD) to Cr in spot samples of urine was shown to vary diurnally in the range of 49% to 157% of the average 24 h value. This finding indicated the need for regular sampling of urine to achieve an accurate average value for the PD: Cr ratio in urine for use in estimating urinary PD excretion and hence microbial protein production in the rumen. It was concluded that continuous, intravenous infusion of CrEDTA resulted in a constant recovery of Cr in the urine of cattle across diets and, provided an intensive sampling regime was followed to account for diurnal variation, it would be suitable as a marker to estimate urine volume and urinary output of PD in grazing cattle.

Linking the populations of barramundi and king threadfin to environmental flows in four rivers of tropical Australia

Linking the populations of barramundi and king threadfin to environmental flows in four rivers of tropical Australia

The diet of the dingo (Canis lupus dingo and hybrids) in north-eastern Australia: a supplement to the paper of Brook and Kutt (2011)

Dingoes and other wild dogs (Canis lupus dingo and hybrids) are generalist predators that consume a wide variety of different prey species within their range. Little is known, however, of the diets of dingoes in north-eastern Australia where the potential for impacts by dingoes exists. Recently new information has been provided on the diets of dingoes from several sites in Queensland, Australia, significantly adding to the body of published knowledge on ecosystems within this region. Further information on the diet of dingoes in north-eastern Australia is added from 1460 scats collected from five sites, representing tropical savannahs, tropical offshore islands (and a matched mainland area), dry sclerophyll forests and peri-urban areas on the fringe of Townsville. Macropods, possums and bandicoots were found to be common prey for dingoes in these areas. Evidence suggested that the frequency of prey remains in scats can be an unreliable indicator of predation risk to potential prey and it was found that novel and unexpected prey species appear in dingo diets as preferred prey become unavailable. The results support the generalisation that dingoes prefer medium- to large-sized native prey species when available but also highlight the capacity for dingoes to exploit populations of both large and small prey species that might not initially be considered at risk from predation based solely on data on scats.

Bacterial Community Structure in the Hindgut of Wild and Captive Dugongs (Dugong dugon)

Dugongs (Dugong dugon) are marine mammals that obtain nutrients through hindgut fermentation of seagrass, however, the microbes responsible have not been identified. This study used denaturing gradient gel electrophoresis (DGGE) and 454-pyrosequencing to profile hindgut bacterial communities in wild dugongs. Faecal samples obtained from 32 wild dugongs representing four size/maturity classes, and two captive dugongs fed on cos lettuce were screened using DGGE. Partial 16S rRNA gene profiles of hindgut bacteria from wild dugong calves and juveniles were grouped together and were different to those in subadults and adults. Marked differences between hindgut bacterial communities of wild and captive dugongs were also observed, except for a single captive whose profile resembled wild adults following an unsuccessful reintroduction to the wild. Pyrosequencing of hindgut communities in two wild dugongs confirmed the stability of bacterial populations, and Firmicutes (average 75.6% of Operational Taxonomic Units [OTUs]) and Bacteroidetes (19.9% of OTUs) dominated. Dominant genera were Roseburia, Clostridium, and Bacteroides. Hindgut microbial composition and diversity in wild dugongs is affected by ontogeny and probably diet. In captive dugongs, the absence of the dominant bacterial DNA bands identified in wild dugongs is probably dependent upon prevailing diet and other captive conditions such as the use of antibiotics. This study represents a first step in the characterisation of a novel microbial ecosystem-the marine hindgut of Sirenia.

The influence of pre- and post-zygotic barriers on interspecific Corymbia hybridization

Corymbia species from different sections hybridize readily, with some of increasing economic importance to plantation forestry. This study explores the locations of reproductive barriers between interspecific Corymbia hybrids and investigates the reproductive success of a wide taxonomic range of C. torelliana hybrid crosses. Pollen, pistil and embryo development were investigated for four C. torelliana crosses (C. torelliana, C. citriodora subsp. citriodora, C. tessellaris and C. intermedia) using fluorescent and standard microscopy to identify the locations of interspecific reproductive isolating barriers. Corymbia torelliana was also crossed with 16 taxa, representing six of the seven Corymbia sections, both Corymbia subgenera and one species each from the related genera, Angophora and Eucalyptus. All crosses were assessed for capsule and seed yields. Interspecific C. torelliana hybridization was controlled by pre-zygotic reproductive isolating barriers inhibiting pollen adhesion to the stigma, pollen germination, pollen tube growth in the style and pollen tube penetration of the micropyle. Corymbia torelliana (subgenus Blakella, sect. Torellianae) was successfully hybridized with Corymbia species from subgenus Blakella, particularly C. citriodora subsp. citriodora, C. citriodora subsp. variegata, C. henryi (sect. Maculatae) and C. tessellaris (sect. Abbreviatae), and subgenus Corymbia, particularly C. clarksoniana and C. erythrophloia (sect. Septentrionales). Attempted intergeneric hybrids between C. torelliana and either Angophora floribunda or Eucalyptus pellita were unsuccessful. Corymbia hybrids were formed between species from different sections and subgenera, but not with species from the related genera Angophora or Eucalyptus. Reproductive isolation between the interspecific Corymbia hybrid crosses was controlled by early- and late-acting pre-zygotic isolating barriers, with reproductive success generally decreasing with increasing taxonomic distance between parent species. These findings support the monophyly of Corymbia and the close relationships of infrageneric clades. The hybridizing propensity of Corymbia species provides opportunities for breeding but suggests risks of environmental gene flow. © The Author 2012. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For Permissions, please email: journals.permissions@oup.com

Effect of Time of Planting and Plant Size on the Productivity of ‘Festival’ and ‘Florida Fortuna’ Strawberry Plants in a Subtropical Environment

The effect of time of planting and plant size on the performance of ‘Festival’ and ‘Florida Fortuna’ strawberry (Fragaria ×ananassa) plants was studied at Nambour in southeastern Queensland, Australia, over 2 years. The main objective of the work was to determine whether small plants yielded proportionally less than large plants as planting was delayed. First, bare-rooted transplants of ‘Festival’ were divided into small (crown diameters ranging from 6 to 10 mm) or large plants (10 to 17 mm) and planted in late March, mid-April, or late April. Second, transplants of ‘Florida Fortuna’ were divided into small (5 to 8 mm) or large plants (8 to 17 mm) and planted in early April, mid-April, or early May. The early planting for each cultivar corresponded with the time that the transplants are first available from commercial strawberry nurseries. Yields were generally greater in plants planted in late March/early April compared with plants planted later. Differences in yield between the small and large plants were consistent across the different times of planting, with the small plants always having lower yields. Small transplants are an issue for the productivity of strawberry fields in this environment whether they are planted early or late. Producers should consider paying a premium for large transplants delivered early in the season.

Immuno-fluorescence staining patterns of leukocyte subsets in the skin of taurine and indicine cattle

The immuno-staining patterns of skin leukocytes were investigated in three breeds of cattle: Holstein–Friesian, Brahman and Santa Gertrudis of similar age before and after tick infestation. The antibodies specific for CD45 and CD45RO reacted with cells in the skin of all Holstein–Friesian cattle but did not react with cells in the skin of any Brahman cattle. The same antibodies reacted with cells from the skin of four (CD45) and seven (CD45RO) of twelve Santa Gertrudis cattle. The antibodies specific for T cells and γδ subset of T cells recognized cells from all three breeds of cattle. The antibody specific for MHC class II molecules labelled cells of mostly irregular shape, presumably dermal dendritic cells and/or macrophages and Langerhans cells. The antibody specific for granulocytes (mAb CH138) reacted with cells only in sections cut from skin with lesions. The antibody specific for CD25+ cells labelled regularly shaped cells that showed a wide range of intensities of staining.

Phytochemicals in Australian papaya cultivars: the effect of maturity and cooking

To measure the effect of maturity and cooking on phytochemical composition and antioxidant capacity of fruit and leaves of four commercially available Australian papaya cultivars (RB1, RB2, RB4 and YB1).