Determination of progesterone directly in milk fat as an improved method for fertility control in cattle

A simple and reliable method for the determination of progesterone in milk fat by radioimmunoassay has been developed. Application of the assay under field conditions has shown that all samples diagnosed " questionable" in the former milk progesterone test (MPT) could be characterized as positive or negative. Thus the total figure of animals daignosed correctly rose from 72.3% to 85.8%. The borderlines (ng progesterone / 10 mcl milk fat) set up for this assay were (1) pregnant and (1) nonpregnant (Day 20) and (.3) in estrus and (.3) not in estrus ( Day 0). Within the application for diagnosis of pregnancy, 81.4% were diagnosed correctly as pregnant while 96.3% were diagnosed correctly as not pregnant. The latter figure is somewhat lower than in the MPT (100%), but under the practical conditions the error lies within the range of false diagnosis due to problems with sample identification.     

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.     

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.     

Flash chromatography on cartridges for the separation of plant extracts: Rules for the selection of chromatographic conditions and comparison with medium pressure liquid chromatography

Empirical rules for the selection of chromatographic conditions on flash chromatography cartridges were developed, with an emphasis on gradient mode. These rules were then tested with separation of extracts from important medicinal plants including Curcuma xanthorrhiza, Piper nigrum and Salvia milthiorrhiza. Sepacore® cartridges enabled a good separation of compounds with a broad range of polarity, as typically found in plant extracts. The chromatographic resolution remained, however, lower than that achieved on classical columns packed with material of smaller particle size. For poorly soluble extracts, solid introduction gave better results than liquid injection.     

Constituents in evening primrose oil with radical scavenging,cyclooxygenase, and neutrophil elastase inhibitory activities

Cold-pressed, non-raffinated evening primrose oil was found to contain lipophilic radical scavengers. A highly enriched fraction of these compounds could be obtained from the oil by extraction with aqueous ethanol and subsequent liquid-liquid partitioning with petroleum. LC-DAD-MS analysis revealed that the fraction contained three aromatic compounds with identical UV and ESI-MS spectra. The compounds were isolated by RP-HPLC and their structures established by chemical and spectroscopic means as 3-O-trans-caffeoyl derivatives of betulinic, morolic, and oleanolic acid. The morolic acid derivative was a new compound. The three esters exhibited pronounced radical scavenging activity against the stable 2,2-diphenyl-1-picrylhydrazyl radical and were potent inhibitors of neutrophil elastase and cyclooxygenase-1 and -2 in vitro. Commercial samples of evening primrose oils contained only traces of these lipophilic antioxidants.     

Toxoplasma gondii prevalence in Israeli crows and Griffon vultures

A cross-sectional Toxoplasma gondii seroprevalence study was performed on free ranging crows (Corvus cornis, Corvus monedula, Corvus splendens) and Griffon vultures (Gyps fulvus) from Israel in order to assess exposure to this pathogen in scavenger birds that feed on animal carcasses and their possible role in the epidemiology of toxoplasmosis. Using the modified agglutination test (MAT) with a cutoff titer of 1:25, 52 of 122 crows (42.6%) and 40 of 101 Griffon vultures (39.6%) were found to be T. gondii seropositive. Crow T. gondii seroprevalence was significantly higher in northern areas of Israel (p = 0.007) where annual precipitation is higher and annual summer maximum temperatures are lower than in the drier and warmer south. Seroprevalence in crows was positively associated with higher human population densities possibly related to the increased cat population in these areas. PCR analysis of brain extracts from crows resulted in the detection of T. gondii DNA in 1 seropositive crow from northern Israel. Genetic analysis of DNA from the positive crow brain confirmed infection with T. gondii type 2 using a multiplex multilocus nested PCR-RFLP (Mn-PCR-RFLP) of the SAG1, 5–3′ SAG2, alt.SAG2, SAG3, BTUB, GRA6, C22-8, c29-2, L358, PK1 and Apico loci. The high T. gondii seroprevalence in these bird species suggests that infected carrion may be responsible for widespread infection of carcass scavenger birds which may further transmit infection to other carnivorous intermediate hosts or feline definitive hosts when consumed post-mortally.     

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.