A framework for evaluation of on-farm mastitis diagnostics in Australia

Numerous culture-based diagnostics are available on the Australian and international markets for on-farm detection of bacterial pathogens in milk. Use of such diagnostics may provide an opportunity to improve the prudent use of antimicrobials in udder health management. Farms are low-resource settings in terms of diagnostic microbiology capacity. The World Health Organisation has identified criteria for the evaluation of diagnostic tests in low resource settings based on Accuracy, Sensitivity, Specificity, User-friendliness, being Rapid or Robust, Equipment-free and being Deliverable (ASSURED). Here, we review how those criteria can be interpreted in the context of microbiological diagnosis of mastitis pathogens, and how on-farm diagnostics that are currently available in Australia perform relative to ASSURED criteria. This evaluation identifies multiple trade-offs, both with regard to scientific criteria and with regards to convenience criteria. More importantly, the purpose of testing may differ between farms, and test performance should be evaluated relative to its intended use. The ability of on-farm mastitis diagnostics to inform mastitis treatment decision-making in a timely and cost-effective manner depends not just on test characteristics but also on farm-specific pathogen prevalence, and on the farm enterprise's priorities and the farm manager's potential courses of action. With most assay evaluations to date conducted in professional laboratories, there is a surprising dearth of information on how well any of the diagnostic tests perform on-farm and, indeed, of the on-farm decision-making processes that they aim to inform.     

Acute cortisol responses of calves to scoop dehorning using local anaesthesia and/or cautery of the wound

Objective To measure plasma cortisol responses in calves dehorned using a scoop after administration of local anaesthesia and/or cautery of the wounds.
Design A physiological study with controls.
Procedure There were six treatments: control handling with and without local anaesthesia, dehorning, dehorning after local anaesthesia, dehorning followed by wound cautery, and dehorning after local anaesthesia followed by wound cautery. Blood samples were taken before and after dehorning.
Results Dehorning caused an increase in plasma cortisol concentrations, which decreased a little to plateau values and then declined to pretreatment values 3 to 4 h after dehorning. The peak was smaller after local anaesthesia was administered but when its effects wore off, cortisol concentrations increased and thereafter were similar to those in the dehorned animals. The combination of local anaesthesia and cautery resulted in a plasma cortisol response similar to those in control calves with or without local anaesthesia.
Conclusions If plasma cortisol concentrations reflect the distress being experienced by the calves, then local anaesthesia reduces the acute distress for about 3 h after dehorning but not during the subsequent 3 to 4 h. Combining local anaesthetic and cautery prevented the significant increase in plasma cortisol following dehorning and may eliminate the acute distress caused by scoop dehorning.     

Incorporating forage grasses in riparian buffers for bioremediation of atrazine, isoxaflutole and nitrate in Missouri

Multi-species tree-shrub-grass riparian buffer systems have been recognized as one of the most cost-effective bioremediation approaches to alleviate nonpoint source agricultural pollution in heavily fertilized systems. However, highly concentrated herbicides in surface and subsurface water and shade cast by trees along the stream bank usually compromise the effectiveness of these systems. Greenhouse trials and field lysimeter studies were conducted to evaluate the tolerance of orchard grass (Dactylis glomerata), smooth bromegrass (Bromus inermis), tall fescue (Festuca arundinacea), timothy (Phleum pratense), and switchgrass (Panicum virgatum) ground covers to atrazine and Balance™ (isoxaflutole) plus their capacity to sequester and degrade these herbicides and their metabolites. Their ability to remove soil nitrate was also quantified. Concentrations of atrazine, Balance™ and their metabolites in the leachate, soil and plant samples were determined by solid phase extraction followed by high performance liquid or gas chromatographic analyses. Distribution of the herbicides and metabolites in the system was calculated using a mass balance approach. Herbicide bioremediation capacity of each lysimeter treatment was determined by the ratio of metabolites to parent herbicide plus metabolites. Bioremediation of nitrate was quantified by comparing nitrate reduction rates in grass treatments to the bare ground control. Based on this herbicide tolerance, bioremediation data and shade tolerance determined in a previous study, it was established that switch grass, tall fescue and smooth bromegrass are good candidates for incorporation into tree-shrub-grass riparian buffer systems designed for the bioremediation of atrazine, Balance™ and nitrate.     

Improved HPLC-MS/MS method for determination of isoxaflutole (balance) and its metabolites in soils and forage plants

A robust multi-residue procedure is needed for the analysis of the pro-herbicide isoxaflutole and its degradates in soil and plant materials at environmentally relevant (<1 microg kg-1) levels. An analytical method using turbo-spray and heat-nebulizer high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was developed for the analysis of isoxaflutole (IXF) and its two metabolites, diketonitrile (DKN) and the benzoic acid metabolite (BA), at sub-microgram per kilogram levels in soil and plant samples. The average recoveries of the three compounds in spiked soil and plant samples ranged from 84 to 110% and 94 to 105%, respectively. The limits of quantification were validated at 0.06 microg kg-1 for soil and 0.3 microg kg-1 for plant samples. The limits of detection (LOD) for soil analysis were 0.01, 0.002, and 0.01 microg kg-1 for IXF, DKN, and BA, respectively. Corresponding LOD for the plant analysis method were 0.05, 0.01, and 0.05 microg kg-1. The developed method was validated using forage grass and soil samples collected from a field lysimeter study in which IXF was applied to each of four forage treatments. Forage plants and soils were sampled for analyses 25 days after IXF application to the soil. In soils, IXF was not detected in any treatment, and DKN was the predominant metabolite found. In forage plants, the concentrations of DKN and BA were 10-100-fold higher than that in soil samples, but IXF was not detected in any forage plants. The much higher proportion of BA to DKN in plant tissues (23-53%), as compared to soils (0-5%), suggested that these forages were capable of detoxifying DKN. The developed methods provided LODs at sub-microgram per kilogram levels to determine the fate of IXF and its metabolites in soils and forage plants, and they also represent considerable improvements in extraction recovery rates and detection sensitivity as compared to previous analytical methods for these compounds.     

Expression of Perilipin 2 (PLIN2) in Porcine Oocytes During Maturation

Perilipins have been reported to limit the interaction of lipases with neutral lipids within the droplets, thereby regulating neutral lipid accumulation and utilization. This study aimed to identify the location and expression of PLIN1 and PLIN2 in porcine oocytes during maturation. Quantitative real‐time polymerase chain reaction (qRT‐PCR), immunostaining and Western blot methods were used to characterize the expression and distribution patterns of PLIN1 and PLIN2 in porcine oocytes. The results showed that PLIN1 was not detectable in porcine oocytes. PLIN2 and BODIPY 493/503‐detected neutral lipid droplets appeared identical distribution patterns and extensive colocalization in both GV and MII porcine oocytes. PLIN2 protein expression was higher in GV oocytes than that in MII oocytes (p < 0.05), although PLIN2 mRNA expression was similar in both groups. These findings suggested that PLIN2 was a major lipid droplet‐associated protein in porcine oocytes.     

Einfluß einer abgestuften Wasserversorgung auf CO2-Assimilation,Transpiration, Substanzproduktion und Wasserverbrauch vonAvena sativa

Zusammenfassung Gefäßkulturen von Hafer in Sand und Erde mit abgestuftem, jeweils gleichbleibend gehaltenem Wasserangebot von 100, 75, 50, 30 und 15% der Boden wasserkapazität ergaben die höchsten Erträge an Gesamttrockenmasse in der Regel bei voller Wassersättigung des Bodens in Verbindung mit gleichzeitig maximalem Wasserverbrauch (bestimmt durch tägliches Wägen der Gefäße). Abnehmende Wasserversorgung verringerte auch Trockenmasse und Wasserverbrauch der Pflanzen. In trockenem Boden und bei reichlicher Wasserzufuhr entwickelte sich die Wurzelmasse im Vergleich mit den oberirdischen Organen stärker als bei mittlerer Bodenfeuchtigkeit. Klimatische Einflüsse, Aussaatzeit und Witterung sowie die Bodenart modifizierten die vom Wasserzustand des Bodens ausgehenden Wirkungen auf Trockenmasseproduktion und Wasserverbrauch, z. B. wurde bei Sommeraussaat gegenüber der Frühjahrsaussaat die Blattentwicklung bedeutend gefördert.Mit registrierenden Infrarot-Gasanalysatoren wurden Tagesgänge von CO₂-Aufnahme, Nachtatmung und Transpiration unter natürlichen Bedingungen gemessen. Bei trockenem Boden war die photosynthetische Leistung der Pflanzen zu Beginn ihrer Entwicklung deutlich gehemmt; sie steigerte sich allmählich und war später merklich größer als bei hohen Wassergaben. Die Transpiration verlief im allgemeinen gleichsinnig wie die Netto-Assimilationsrate, wurde jedoch auch nachts meist auf einer gewissen Höhe fortgesetzt. Die Nachtatmung der sehr trocken gehaltenen Pflanzen war gegenüber den anderen Wasserstufen meist erhöht. Die Trockenkulturen zeigten einen größeren Lichtbedarf; der Lichtkompensationspunkt wurde gewöhnlich erst bei höherer Beleuchtungsstärke erreicht als bei guter Wasserversorgung. Daraus wird auf erhöhten Energiebedarf der sehr trocken kultivierten Pflanzen geschlossen. Das Verhältnis zwischen CO₂-Aufnahme und Transpiration wurde durch unterschiedliche Bodenwasserversorgung meist nur wenig beeinflußt. Die Ergebnisse der CO₂-Assimilationsmessungen stehen im großen und ganzen mit dem wöchentlich festgestellten Trockensubstanzgewinn in Einklang.

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Summary Cultures ofAvena sativa L. grown in pots with sand or garden soil with a graduated, in each case fairly constant water supply of 100, 75, 50, 30, and 15% of field capacity yielded maximum total dry weight, as a rule, in fully water-saturated soils combined with maximum water consumption (measured by daily weighing). Decreasing water supply reduced dry matter production and water consumption of plants. In dry soil and at high degrees of water saturation roots developed more dry matter in comparison to the shoots than at medium soil moisture. Results in dry matter production and water consumption, as caused by water relations in soil, are modified additionally by climatic factors, sowing date and weather, and soil type, e.g. sowing in summer considerably favoured leaf development, compared with the spring seed.Daily courses of CO₂-uptake, night respiration and transpiration were registered by infrared-gasanalyzers. In very dry soil photosynthetic activity of plants was distinctly hampered during the first weeks of development; it gradually increased, and at a later stage photosynthetic activity was markedly higher than at abundant water supply. In general transpiration followed the net assimilation rate, it was continued, however, even over night on a certain level. Night respiration of plants growing in dry soil was higher in relation to the other water stages. Those plants, too, required more light for dry matter production; their light compensation point arrived at higher light intensity than under abundant water supply. This behaviour seems to indicate higher energy requirement of plants distinctly influenced by water stress. Relations between CO₂-uptake and transpiration were but slightly influenced by different soil water level. Results of gas exchange measurements on CO₂-assimilation agreed, as a rule, with the weekly dry matter measurements by weighting.

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Role of Luteinizing Hormone in Primiparous Sow Responses to Split Weaning

In 45 primiparous sows, we examined endocrine, ovarian and reproductive responses to split-weaning or five injections per day of 800 ng GnRH from 18 to 21 days of lactation. There was no effect of treatment on absolute or changes in sow weight or backfat depth during lactation. Average piglet growth rates were similar among treatments except that piglets suckling split-weaned sows grew faster (p < 0.05) during days 18–21. On day 18, mean plasma LH concentrations and LH pulse frequency remained relatively stable in conventionally weaned sows but increased (p < 0.01) in response to split-weaning and GnRH. Prior to weaning on day 21, mean plasma LH concentrations remained elevated in GnRH-treated sows but had returned to control levels in split weaned sows. There was no treatment effect on preweaning LH pulse frequency noted on day 21. Weaning was associated with an increase in plasma LH concentrations in all the treatment groups. Mean plasma IGF-I remained relatively constant in conventionally weaned and GnRH sows, decreased in response to split weaning on day 18 (p < 0.02), but were elevated (p < 0.03) in split wean sows on day 21. On the day after weaning, split wean sows had more (p < 0.04) ovarian follicles ≥3 mm than conventionally weaned sows, with GnRH sows being intermediate. The wean-to-oestrus interval was reduced in split-wean sows compared with those conventionally weaned (p < 0.01), with GnRH sows being intermediate. There was no effect of treatment on ovulation rates, numbers of embryos, or embryonic survival rates. These data indicate that split-weaning of litters results in a more rapid return to oestrus after weaning and that this effect is associated with a transient acute increase in circulating gonadotrophins and earlier resumption of ovarian follicular development.     

Elevating glucose and insulin secretion by carbohydrate formulation diets in late lactation to improve post‐weaning fertility in primiparous sows

Primiparous (P1) sows commonly lose excessive body reserves to meet energy requirements for maintenance and milk production during lactation, and consequently, post‐weaning reproductive performance may be compromised. The present studies determined whether ad libitum feeding a glucogenic carbohydrate diet (CHO) during late lactation could stimulate insulin and glucose secretion (experiment 1) and improve subsequent litter size (experiment 2). For experiment 1, 15 P1 sows, and for experiment 2, 99 P1 sows (198.5 ± 2.7 kg) were allocated randomly according to suckled litter size (≥10 piglets), either to a CHO diet (14.3 MJ DE/kg, 19.8% crude protein) or a standard lactation diet (control; 14.2 DE MJ/kg, 19.5% crude protein) at 8 days before weaning. The CHO diet aimed to provide glucogenic content (extruded wheat, dextrose and sugar) as energy sources instead of fat sources without changing total dietary energy. Pre‐prandial plasma glucose and insulin concentrations were not influenced by treatments. However, post‐prandial plasma glucose and insulin concentrations and their peaks were both higher (p < .05) compared to the control treatment. Body weight loss during lactation was relatively low at 3%–4% for both treatments and did not differ between control and CHO treatments (?7.6 ± 1.6 vs ?5.4 ± 1.2 kg; p > .05). Second litter size was not influenced by diet (p > .05), but the weaning‐to‐mating interval was shorter in CHO sows (p < .05). This study demonstrates that providing an enriched CHO diet in late lactation did influence post‐weaning follicle growth but did not improve subsequent litter size. This may be due to the primiparous sows in this study not experiencing severe negative energy balance and there was no second litter syndrome in this farm which limited the ability of diet to improve sow fertility.     

Changing trends in mastitis

The global dairy industry, the predominant pathogens causing mastitis, our understanding of mastitis pathogens and the host response to intramammary infection are changing rapidly. This paper aims to discuss changes in each of these aspects. Globalisation, energy demands, human population growth and climate change all affect the dairy industry. In many western countries, control programs for contagious mastitis have been in place for decades, resulting in a decrease in occurrence of Streptococcus agalactiae and Staphylococcus aureus mastitis and an increase in the relative impact of Streptococcus uberis and Escherichia coli mastitis. In some countries, Klebsiella spp. or Streptococcus dysgalactiae are appearing as important causes of mastitis. Differences between countries in legislation, veterinary and laboratory services and farmers' management practices affect the distribution and impact of mastitis pathogens. For pathogens that have traditionally been categorised as contagious, strain adaptation to human and bovine hosts has been recognised. For pathogens that are often categorised as environmental, strains causing transient and chronic infections are distinguished. The genetic basis underlying host adaptation and mechanisms of infection is being unravelled. Genomic information on pathogens and their hosts and improved knowledge of the host's innate and acquired immune responses to intramammary infections provide opportunities to expand our understanding of bovine mastitis. These developments will undoubtedly contribute to novel approaches to mastitis diagnostics and control.