Ruminococcus bromii, identification and isolation as a dominant community member in the rumen of cattle fed a barley diet

Aims: To identify dominant bacteria in grain (barley)-fed cattle for isolation and future use to increase the efficiency of starch utilization in these cattle. Methods and Results: Total DNA was extracted from samples of the rumen contents from eight steers fed a barley diet for 9 and 14 days. Bacterial profiles were obtained using denaturing gradient gel electrophoresis (DGGE) of the PCR-amplified V2/V3 region of the 16S rRNA genes from total bacterial DNA. Apparently dominant bands were excised and cloned, and the clone insert sequence was determined. One of the most common and dominant bacteria present was identified as Ruminococcus bromii. This species was subsequently isolated using traditional culture-based techniques and its dominance in the grain-fed cattle was confirmed using a real-time Taq nuclease assay (TNA) designed for this purpose. In some animals, the population of R. bromii reached densities above 1010R. bromii cell equivalents per ml or approximately 10% of the total bacterial population. Conclusions: Ruminococcus bromii is a dominant bacterial population in the rumen of cattle fed a barley-based diet. Significance and Impact of the Study: Ruminococcus bromii YE282 may be useful as a probiotic inoculant to increase the efficiency of starch utilization in barley-fed cattle. The combination of DGGE and real-time TNA has been an effective process for identifying and targeting for isolation, dominant bacteria in a complex ecosystem.

A Comparison of Methods for Describing Irregular Animal Growth and Testing for Treatment Effects

In many designed experiments with animals liveweight is recorded several times during the trial. Such data are commonly referred to as repeated measures data. An aim of such experiments is generally to compare the growth patterns for the applied treatments. This paper discusses some of the methods of analysing repeated measures data and illustrates the use of cubic smoothing splines to describe irregular cattle growth data. Animal production for a consuming world : proceedings of 9th Congress of the Asian-Australasian Association of Animal Production Societies [AAAP] and 23rd Biennial Conference of the Australian Society of Animal Production [ASAP] and 17th Annual Symposium of the University of Sydney, Dairy Research Foundation, [DRF]. 2-7 July 2000, Sydney, Australia.

Low-density koala (Phascolarctos cinereus) populations in the mulgalands of south-west Queensland. II. Distribution and diet

This study used faecal pellets to investigate the broadscale distribution and diet of koalas in the mulgalands biogeographic region of south-west Queensland. Koala distribution was determined by conducting faecal pellet searches within a 30-cm radius of the base of eucalypts on 149 belt transects, located using a multi-scaled stratified sampling design. Cuticular analysis of pellets collected ffom 22 of these sites was conducted to identify the dietary composition of koalas within the region. Our data suggest that koala distribution is concentrated in the northern and more easterly regions of the study area, and appears to be strongly linked with annual rainfall. Over 50% of our koala records were obtained from non-riverine communities, indicating that koalas in the study area are not primarily restricted to riverine communities, as bas frequently been suggested. Cuticular analysis indicates that more than 90% of koala diet within the region consists of five eucalypt species. Our data highlights the importance of residual Tertiary landforms to koala conservation in the region.

Using simulation techniques to investigate methods to determine resistance of helminths to anthelmintic treatment

The widespread and increasing resistance of internal parasites to anthelmintic control is a serious problem for the Australian sheep and wool industry. As part of control programmes, laboratories use the Faecal Egg Count Reduction Test (FECRT) to determine resistance to anthelmintics. It is important to have confidence in the measure of resistance, not only for the producer planning a drenching programme but also for companies investigating the efficacy of their products. The determination of resistance and corresponding confidence limits as given in anthelmintic efficacy guidelines of the Standing Committee on Agriculture (SCA) is based on a number of assumptions. This study evaluated the appropriateness of these assumptions for typical data and compared the effectiveness of the standard FECRT procedure with the effectiveness of alternative procedures. Several sets of historical experimental data from sheep and goats were analysed to determine that a negative binomial distribution was a more appropriate distribution to describe pre-treatment helminth egg counts in faeces than a normal distribution. Simulated egg counts for control animals were generated stochastically from negative binomial distributions and those for treated animals from negative binomial and binomial distributions. Three methods for determining resistance when percent reduction is based on arithmetic means were applied. The first was that advocated in the SCA guidelines, the second similar to the first but basing the variance estimates on negative binomial distributions, and the third using Wadley’s method with the distribution of the response variate assumed negative binomial and a logit link transformation. These were also compared with a fourth method recommended by the International Co-operation on Harmonisation of Technical Requirements for Registration of Veterinary Medicinal Products (VICH) programme, in which percent reduction is based on the geometric means. A wide selection of parameters was investigated and for each set 1000 simulations run. Percent reduction and confidence limits were then calculated for the methods, together with the number of times in each set of 1000 simulations the theoretical percent reduction fell within the estimated confidence limits and the number of times resistance would have been said to occur. These simulations provide the basis for setting conditions under which the methods could be recommended. The authors show that given the distribution of helminth egg counts found in Queensland flocks, the method based on arithmetic not geometric means should be used and suggest that resistance be redefined as occurring when the upper level of percent reduction is less than 95%. At least ten animals per group are required in most circumstances, though even 20 may be insufficient where effectiveness of the product is close to the cut off point for defining resistance.

Investigation and application of methods for enumerating heterotrophs and Escherichia coli in the air within piggery sheds

Aims: To investigate methods for the recovery of airborne bacteria within pig sheds and to then use the appropriate methods to determine the levels of heterotrophs and Escherichia coli in the air within sheds. Methods and Results: AGI-30 impingers and a six-stage Andersen multi-stage sampler (AMS) were used for the collection of aerosols. Betaine and catalase were added to impinger collection fluid and the agar plates used in the AMS. Suitable media for enumerating E. coli with the Andersen sampler were also evaluated. The addition of betaine and catalase gave no marked increase in the recovery of heterotrophs or E. coli. No marked differences were found in the media used for enumeration of E. coli. The levels of heterotrophs and E. coli in three piggeries, during normal pig activities, were 2Æ2 · 105 and 21 CFU m)3 respectively. Conclusions: The failure of the additives to improve the recovery of either heterotrophs or E. coli suggests that these organisms are not stressed in the piggery environment. The levels of heterotrophs in the air inside the three Queensland piggeries investigated are consistent with those previously reported in other studies. Flushing with ponded effluent had no marked or consistent effect on the heterotroph or E. coli levels. Significance and Impact of the Study: Our work suggests that levels of airborne heterotrophs and E. coli inside pig sheds have no strong link with effluent flushing. It would seem unlikely that any single management activity within a pig shed has a dominant influence on levels of airborne heterotrophs and E. coli

Aspects of reproduction and diet of the Australian endemic skate Dipturus polyommata (Ogilby) (Elasmobranchii: Rajidae), by-catch of a commercial prawn trawl fishery

The Australian endemic skate Dipturus polyommata collected from by-catch of a benthic prawn fishery off southern Queensland was examined to provide information on reproduction and diet. Morphological relationships of total length (LT) to disc width and LT to mass were estimated. Size at birth was estimated at c. 100-110 mm and size at first feeding at c. 105-110 mm LT. Size at 50% maturity (LT50 and 95% CI) was 321 (305-332) and 300 (285-306) mm LT for females and males, respectively. Size at first maturity corresponded to 87.7% of observed maximum size in females (366 mm LT) and 87.5% in males (343 mm L T). Two females, representing 18.2% of mature females sampled in the austral winter were each carrying two egg cases. Descriptions of egg cases are given. Diet described by the index of relative importance as a percentage (%IRI) was predominantly crustacean based with carid shrimps (53.64%) and penaeoid prawns (23.30%) the most significant prey groups. Teleosts (11.72%), gammarid amphipods (5.31%) and mysids (4.72%) were also important to the diet of the species, while a further six prey groups made only a minor contribution to diet (1.31%). An ontogenetic change was evident between the diets of immature and mature skates. Immature animals fed more extensively on carids and amphipods and mature animals on penaeoids, teleosts and mysids.

Comparison of sampling sites and detection methods for Haemophilus parasuis

Objective To improve the isolation rate and identification procedures for Haemophilus parasuis from pig tissues. Design Thirteen sampling sites and up to three methods were used to confirm the presence of H. parasuis in pigs after experimental challenge. Procedure Colostrum-deprived, naturally farrowed pigs were challenged intratracheally with H parasuis serovar 12 or 4. Samples taken during necropsy were either inoculated onto culture plates, processed directly for PCR or enriched prior to being processed for PCR. The recovery of H parasuis from different sampling sites and using different sampling methods was compared for each serovar. Results H parasuis was recovered from several sample sites for all serovar 12 challenged pigs, while the trachea was the only positive site for all pigs following serovar 4 challenge. The method of solid medium culture of swabs, and confirmation of the identity of cultured bacteria by PCR, resulted in 38% and 14% more positive results on a site basis for serovars 12 and 4, retrospectively, than direct PCR on the swabs. This difference was significant in the serovar 12 challenge. Conclusion Conventional culture proved to be more effective in detecting H parasuis than direct PCR or PCR on enrichment broths. For subacute (serovar 4) infections, the most successful sites for culture or direct PCR were pleural fluid, peritoneal fibrin and fluid, lung and pericardial fluid. For acute (serovar 12) infections, the best sites were lung, heart blood, affected joints and brain. The methodologies and key sampling sites identified in this study will enable improved isolation of H parasuis and aid the diagnosis of Glässer's disease.

A comparison of mark-recapture distance-sampling methods applied to aerial surveys of eastern grey kangaroos

Aerial surveys of kangaroos (Macropus spp.) in Queensland are used to make economically important judgements on the levels of viable commercial harvest. Previous analysis methods for aerial kangaroo surveys have used both mark-recapture methodologies and conventional distance-sampling analyses. Conventional distance sampling has the disadvantage that detection is assumed to be perfect on the transect line, while mark-recapture methods are notoriously sensitive to problems with unmodelled heterogeneity in capture probabilities. We introduce three methodologies for combining together mark-recapture and distance-sampling data, aimed at exploiting the strengths of both methodologies and overcoming the weaknesses. Of these methods, two are based on the assumption of full independence between observers in the mark-recapture component, and this appears to introduce more bias in density estimation than it resolves through allowing uncertain trackline detection. Both of these methods give lower density estimates than conventional distance sampling, indicating a clear failure of the independence assumption. The third method, termed point independence, appears to perform very well, giving credible density estimates and good properties in terms of goodness-of-fit and percentage coefficient of variation. Estimated densities of eastern grey kangaroos range from 21 to 36 individuals km-2, with estimated coefficients of variation between 11% and 14% and estimated trackline detection probabilities primarily between 0.7 and 0.9.

Diet quality and liveweight gain of steers grazing Leucaena-grass pasture estimated with faecal near infrared reflectance spectroscopy (F.NIRS)

Three drafts of Bos indicus cross steers (initially 178-216 kg) grazed Leucaena-grass pasture [Leucaena leucocephala subspecies glabrata cv. Cunningham with green panic (Panicum maximum cv. trichoglume)] from late winter through to autumn during three consecutive years in the Burnett region of south-east Queensland. Measured daily weight gain (DWGActual) of the steers was generally 0.7-1.1 kg/day during the summer months. Estimated intakes of metabolisable energy and dry matter (DM) were calculated from feeding standards as the intakes required by the steers to grow at the DWGActual. Diet attributes were predicted from near infrared reflectance spectroscopy spectra of faeces (F.NIRS) using established calibration equations appropriate for northern Australian forages. Inclusion of some additional reference samples from cattle consuming Leucaena diets into F.NIRS calibrations based on grass and herbaceous legume-grass pastures improved prediction of the proportion of Leucaena in the diet. Mahalanobis distance values supported the hypothesis that the F.NIRS predictions of diet crude protein concentration and DM digestibility (DMD) were acceptable. F.NIRS indicated that the percentage of Leucaena in the diet varied widely (10-99%). Diet crude protein concentration and DMD were usually high, averaging 12.4 and 62%, respectively, and were related asymptotically to the percentage of Leucaena in the diet (R2 = 0.48 and 0.33, respectively). F.NIRS calibrations for DWG were not satisfactory to predict this variable from an individual faecal sample since the s.e. of prediction were 0.33-0.40 kg/day. Cumulative steer liveweight (LW) predicted from F.NIRS DWG calibrations, which had been previously developed with tropical grass and grass-herbaceous legume pastures, greatly overestimated the measured steer LW; therefore, these calibrations were not useful. Cumulative steer LW predicted from a modified F.NIRS DWG calibration, which included data from the present study, was strongly correlated (R2 = 0.95) with steer LW but overestimated LW by 19-31 kg after 8 months. Additional reference data are needed to develop robust F.NIRS calibrations to encompass the diversity of Leucaena pastures of northern Australia. In conclusion, the experiment demonstrated that F.NIRS could improve understanding of diet quality and nutrient intake of cattle grazing Leucaena-grass pasture, and the relationships between nutrient supply and cattle growth.

Comparison of nitrogen fertilization methods and rates for subsurface drip irrigated corn in the semi-arid Great Plains

In semi-arid areas such as western Nebraska, interest in subsurface drip irrigation (SDI) for corn is increasing due to restricted irrigation allocations. However, crop response quantification to nitrogen (N) applications with SDI and the environmental benefits of multiple in-season (IS) SDI N applications instead of a single early-season (ES) surface application are lacking. The study was conducted in 2004, 2005, and 2006 at the University of Nebraska-Lincoln West Central Research and Extension Center in North Platte, Nebraska, comparing two N application methods (IS and ES) and three N rates (128, 186, and 278 kg N ha(-1)) using a randomized complete block design with four replications. No grain yield or biomass response was observed in 2004. In 2005 and 2006, corn grain yield and biomass production increased with increasing N rates, and the IS treatment increased grain yield, total N uptake, and gross return after N application costs (GRN) compared to the ES treatment. Chlorophyll meter readings taken at the R3 corn growth stage in 2006 showed that less N was supplied to the plant with ES compared to the IS treatment. At the end of the study, soil NO3-N masses in the 0.9 to 1.8 m depth were greater under the IS treatment compared to the ES treatment. Results suggested that greater losses of NO3-N below the root zone under the ES treatment may have had a negative effect on corn production. Under SDI systems, fertigating a recommended N rate at various corn growth stages can increase yields, GRN, and reduce NO3-N leaching in soils compared to concentrated early-season applications.

Cannibalism contributes significantly to the diet of cultured sand crabs, Portunus pelagicus (L.): A dual stable isotope study

The significance of cannibalism in the diet of juvenile pond-cultured blue swimmer crabs (Portunus pelagicus (L.)) was investigated using dual stable isotope analysis of carbon and nitrogen. In a laboratory feeding experiment, δ15N demonstrated a constant trophic shift (Δδ15N ≈+ 1.6‰), and therefore seemed to be a reliable indicator for assessing trophic position for P. pelagicus. This agrees with previously reported trends. Difference in growth rate did not seem to influence δ15N values. In contrast, δ13C did not display consistent shifts between trophic levels (range of Δδ13C: + 1 to + 1.7‰). The results from the pond experiment showed that larger individuals had a more enriched δ15N than smaller individuals, which, when compared to the results from the laboratory experiment, indicates that larger individuals were at a higher trophic level. This is most likely due to cannibalism prevailing in the pond rather than a direct result of faster growth rate. Cannibalistic behaviour might further increase growth, resulting in the observed positive correlation between size and δ15N.

Evaluation of partial replacement of live and fresh feeds with a formulated diet and the influence of weaning Panulirus ornatus phyllosomata onto a formulated diet during early ontogeny

We have evaluated the potential of a formulated diet as a replacement for live and fresh feeds for 7-day post-hatch Panulirus ornatus phyllosomata and also investigated the effect of conditioning phyllosomata for 14-21 days on live feeds prior to weaning onto a 100% formulated diet. In the first trial, the highest survival (>55%) was consistently shown by phyllosomata fed a diet consisting of a 50% combination of Artemia nauplii and 50% Greenshell mussel, followed by phyllosomata fed 50% Artemia nauplii and 50% formulated diet and, thirdly, by those receiving 100% Artemia nauplii. The second trial assessed the replacement of on-grown Artemia with proportions of formulated diet and Greenshell mussel that differed from those used in trial 1. Phyllosomata fed a 75% combination of formulated diet and 25% on-grown Artemia and 50% on-grown Artemia and 50% Greenshell mussel consistently showed the highest survival (>75%). Combinations of Greenshell mussel and formulated diet resulted in significantly (P < 0.05) reduced survival. In trial 3, phyllosomata were conditioned for 14, 18 or 21 days on Artemia nauplii prior to weaning onto a 100% formulated diet, which resulted in survival rates that were negatively related to the duration of feeding Artemia nauplii. In the final trial, phyllosomata were conditioned for 14 days on live on-grown Artemia prior to weaning onto one of three formulated diets (one diet with 44% CP and two diets with 50%). Phyllosomata fed a 44% CP diet consistently showed the highest survival (>35%) among all treatments, while those fed a 50%-squid CP diet showed a significant (P < 0.05) increase in mortality at day 24. The results of these trials demonstrate that hatcheries can potentially replace 75% of live on-grown Artemia with a formulated diet 7 days after hatch. The poor performance associated with feeding combinations of Greenshell mussel and formulated diet, and 100% formulated diet as well as conditioning phyllosomata for 14-21 days on live feeds prior to weaning onto a formulated diet highlights the importance of providing Artemia to stimulate feeding.

Effects of diet, temperature and photoperiod on development and survival of the bigeyed bug, Geocoris lubra

Bigeyed bugs (Geocoris spp., Hemiptera: Geocoridae) are common predators in Australian agricultural crops yet the development and reproductive biology of Australian geocorids has not been described before. Here we present the effects of diet, temperature and photoperiod on the development and survival of Geocoris lubra Kirkaldy from egg to adult. Nymphal survival of G. lubra reared on live aphids (Aphis gossypii Glover) was very low but improved slightly on a diet of Helicoverpa armigera (Hübner) eggs. Development was faster and nymphal survival improved significantly at 27°C compared with 25°C. Further investigation at 27°C showed photoperiod influenced development time, but not survival of immature G. lubra. Development time was significantly longer at 10L:14D. Fecundity of first generation G. lubra was not affected by photoperiod, although egg viability was greater at 12L:12D.

Hardness Methods for Testing Maize Kernels

Maize is a highly important crop to many countries around the world, through the sale of the maize crop to domestic processors and subsequent production of maize products and also provides a staple food to subsistance farms in undeveloped countries. In many countries, there have been long-term research efforts to develop a suitable hardness method that could assist the maize industry in improving efficiency in processing as well as possibly providing a quality specification for maize growers, which could attract a premium. This paper focuses specifically on hardness and reviews a number of methodologies as well as important biochemical aspects of maize that contribute to maize hardness used internationally. Numerous foods are produced from maize, and hardness has been described as having an impact on food quality. However, the basis of hardness and measurement of hardness are very general and would apply to any use of maize from any country. From the published literature, it would appear that one of the simpler methods used to measure hardness is a grinding step followed by a sieving step, using multiple sieve sizes. This would allow the range in hardness within a sample as well as average particle size and/or coarse/fine ratio to be calculated. Any of these parameters could easily be used as reference values for the development of near-infrared (NIR) spectroscopy calibrations. The development of precise NIR calibrations will provide an excellent tool for breeders, handlers, and processors to deliver specific cultivars in the case of growers and bulk loads in the case of handlers, thereby ensuring the most efficient use of maize by domestic and international processors. This paper also considers previous research describing the biochemical aspects of maize that have been related to maize hardness. Both starch and protein affect hardness, with most research focusing on the storage proteins (zeins). Both the content and composition of the zein fractions affect hardness. Genotypes and growing environment influence the final protein and starch content and. to a lesser extent, composition. However, hardness is a highly heritable trait and, hence, when a desirable level of hardness is finally agreed upon, the breeders will quickly be able to produce material with the hardness levels required by the industry.

Seasonal changes in pasture quality and diet selection and their relationship with liveweight gain of steers grazing tropical grass and grass-legume pastures in northern Australia.

The variation in liveweight gain in grazing beef cattle as influenced by pasture type, season and year effects has important economic implications for mixed crop-livestock systems and the ability to better predict such variation would benefit beef producers by providing a guide for decision making. To identify key determinants of liveweight change of Brahman-cross steers grazing subtropical pastures, measurements of pasture quality and quantity, and diet quality in parallel with liveweight were made over two consecutive grazing seasons (48 and 46 weeks, respectively), on mixed Clitoria ternatea/grass, Stylosanthes seabrana/grass and grass swards (grass being a mixture of Bothriochloa insculpta cv. Bisset, Dichanthium sericeum and Panicum maximum var. trichoglume cv. Petrie). Steers grazing the legume-based pastures had the highest growth rate and gained between 64 and 142 kg more than those grazing the grass pastures in under 12 months. Using an exponential model, green leaf mass, green leaf %, adjusted green leaf % (adjusted for inedible woody legume stems), faecal near infrared reflectance spectroscopy predictions of diet crude protein and diet dry matter digestibility, accounted for 77, 74, 80, 63 and 60%, respectively, of the variation in daily weight gain when data were pooled across pasture types and grazing seasons. The standard error of the regressions indicated that 95% prediction intervals were large (+/- 0.42-0.64 kg/head.day) suggesting that derived regression relationships have limited practical application for accurately estimating growth rate. In this study, animal factors, especially compensatory growth effects, appeared to have a major influence on growth rate in relation to pasture and diet attributes. It was concluded that predictions of growth rate based only on pasture or diet attributes are unlikely to be accurate or reliable. Nevertheless, key pasture attributes such as green leaf mass and green leaf% provide a robust indication of what proportion of the potential growth rate of the grazing animals can be achieved.