Immunological detection of cardiac glycosides in plants

Summary Australian native and introduced plants were examined, using digoxin immunoassays, to detect the presence of cross-reacting cardiac glycosides. These compounds were found in 27 species from 20 genera. The assay technique can also be used on serum samples to confirm cardiac glycoside ingestion.     

Anthelmintic resistance in Australian sheep nematode populations

SUMMARY The resistance status of gastro–intestinal nematodes to anthelmintics was evaluated on 881 sheep farms throughout Australia during 1991–92. Resistance was shown to be widespread. Overall, 85% of farms had sheep infected with nematodes resistant to benzimidazole, 65% to levamisole and 34% to combination (benzimidazole + levamisole) products. Resistance to ivermectin was not detcted. On only 9% of farms did all anthelmintic groups reduce egg counts by greater than or equal to 95%. The culture of faeces from untreated sheep showed Telodorsagia circumcincta, Trichostrongylus spp, Chabertia ovina and Haemonchus contortus to be the principal species. The nature and prevalence of resistance was not significantly correlated with stocking rate. However, resistance to combination products was almost twice as prevalent on farms in areas with an average annual rainfall of greater than 500 mm.     

Further investigation of lead exposure as a potential threatening process for a scavenging marsupial species

There is a growing recognition of the harmful effects of lead exposure on avian and mammalian scavengers. This can lead to both lethal and non-lethal effects which may negatively impact wildlife populations. Our objective was to assess medium-term lead exposure in wild Tasmanian devils (Sarcophilus harrisii). Frozen liver samples (n = 41), opportunistically collected in 2017–2022, were analysed using inductively coupled plasma mass spectrometry (ICP-MS) to determine liver lead concentrations. These results were then used to calculate the proportion of animals with elevated lead levels (>5 mg/kg dry weight) and examine the role of explanatory variables that may have influenced the results. The majority of samples analysed were from the south-east corner of Tasmania, within 50 km of Hobart. No Tasmanian devil samples were found to have elevated lead levels. The median liver lead concentration was 0.17 mg/kg (range 0.05–1.32 mg/kg). Female devils were found to have significantly higher liver lead concentrations than males (P = 0.013), which was likely related to lactation, but other variables (age, location, body mass) were not significant. These results suggest that wild Tasmanian devil populations currently show minimal medium-term evidence of exposure to lead pollution, although samples were concentrated in peri-urban areas. The results provide a baseline level which can be used to assess the impact of any future changes in lead use in Tasmania. Furthermore, these data can be used as a comparison for lead exposure studies in other mammalian scavengers, including other carnivorous marsupial species.     

Derivation of Canine Induced Pluripotent Stem Cells

Dogs and humans have many inherited genetic diseases in common and conditions that are increasingly prevalent in humans also occur naturally in dogs. The use of dogs for the experimental and clinical testing of stem cell and regenerative medicine products would benefit canine health and welfare and provide relevant animal models for the translation of therapies to the human field. Induced pluripotent stem cells (iPSCs) have the capacity to turn into all cells of the body and therefore have the potential to provide cells for therapeutic use and for disease modelling. The objective of this study was to derive and characterize iPSCs from karyotypically abnormal adult canine cells. Aneuploid adipose‐derived mesenchymal stromal cells (AdMSCs) from an adult female Weimeraner were re‐programmed into iPSCs via overexpression of four human pluripotency factors (Oct 4, Sox2, Klf4 and c‐myc) using retroviral vectors. The iPSCs showed similarity to human ESCs with regard to morphology, pluripotency marker expression and the ability to differentiate into derivatives of all three germ layers in vitro (endoderm, ectoderm and mesoderm). The iPSCs also demonstrated silencing of the viral transgenes and re‐activation of the silent X chromosome, suggesting full reprogramming had occurred. The levels of aneuploidy observed in the AdMSCs were maintained in the iPSCs. This finding demonstrates the potential for generating canine induced pluripotent stem cells for use as disease models in addition to regenerative medicine and pharmaceutical testing.     

Target region amplification polymorphism (TRAP) for assessing genetic diversity and marker-trait associations in chickpea (Cicer arietinum L.) germplasm

The effective utilization of crop diversity held in genebanks depends on our knowledge of useful traits and available markers associated with the target traits. Target region amplification polymorphism (TRAP) was used to evaluate the genetic diversity and underlying relationships among 263 accessions of chickpea landraces maintained by the USDA-ARS Western Regional Plant Introduction Station in Pullman, WA, USA. Two-hundred sixty-two TRAP markers were amplified by eight primer combinations. Altogether, 110 (42 %) markers were polymorphic, the other 152 (58 %) displayed no variation. These polymorphic markers revealed important differences among the accessions, with an estimated, mean pair-wise genetic distance of 25.82 %, ranging from 2.8 to 50.0 %. Genetic distance analysis divided the accessions into two major groups, with 113 and 150 accessions each, and substantial association between molecular diversity and geographic origin was evident. Bayesian analysis of population structure revealed two groups (K = 2) with evidence for six sub-groups. Additionally, the population structure of a subset of 110 lines was determined (K = 3) for testing marker-trait associations (MTAs). Phenotypic traits included the concentrations of protein and nine mineral elements in the seeds. Two MTAs were significant (p < 0.01) for concentrations of Ca and K, and three MTAs were significant for Cu and Ni concentrations. The results indicate that this population is useful for genome-wide association studies on other economic traits given the level of genetic diversity uncovered and the marker-trait associations in seed minerals discovered.     

Estimating the mass of asteroid 253 mathilde from tracking data during the NEAR flyby

The terminal navigation of the Near Earth Asteroid Rendezvous (NEAR) spacecraft during its close flyby of asteroid 253 Mathilde involved coordinated efforts to determine the heliocentric orbits of the spacecraft and Mathilde and then to determine the relative trajectory of the spacecraft with respect to Mathilde. The gravitational perturbation of Mathilde on the passing spacecraft was apparent in the spacecraft tracking data. As a result of the accurate targeting achieved, these data could be used to determine Mathilde's mass as 1.033 (+/- 0.044) x 10(20) grams. Coupled with a volume estimate provided by the NEAR imaging team, this mass suggests a low bulk density for Mathilde of 1.3 grams per cubic centimeter.     

Wound healing in potato tuber tissue. 1. Effects of maturity at harvest

Summary Potatoes of cvs Record (maincrop) and Red Craigs Royal (second early) were harvested at intervals from 22 August to 14 October. Half of the plots were mechanically defoliated on 22 August, by which date haulms of cv. Red Craigs Royal were largely dead. Tubers of cv. Record were immature at the first lifting date and matured thereafter, particularly after defoliation. Wound healing rates were assessed from the ability of discs of tuber tissue, cut the day after harvest, to become resistant to the loss of water vapour as suberization progressed at high relative humidity. Cv. Red Craigs Royal showed little variation in wound healing rate. Wound healing in cv. Record became more rapid with increasing maturity, particularly after defoliation when it reached a maximum by 1 October. After storage until March in the following year, the effect of maturity at lifting on wound healing rate was no longer evident.

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Zusammenfassung Zweck dieser Versuche war herauszufinden ob die Intensit?t der Wundheilung unmittelbar nach der Ernte bei unreifen Knollen beeintr?chtigt war und ob irgendwelche Effekte nach Lagerung erhalten blieben. Knollen der mittelfrühen Sorte Red Craigs Royal und der sp?ten Sorte Record wurden um 22. August bis zum 14. Oktober 1979 in Intervallen geerntet. Am 22. August wurde die H?lfte der Parzellen mechanisch abget?tet, der Rest starb auf natürliche Weise ab. Das natürliche Absterben war bei der Sorte Red Craigs Royal am 22. August und bei Record am l. Oktober fast komplett. Am Tage nach jeder Ernte wurden 11×4 mm starke Scheiben aus dem Mark entnommen, nach 10 Tagen die Entwicklung der internen Widerstandsf?higkeit gegen Wasserverlust durchgeführt. Diese Massnahme stellt ein Mass der Suberinisierung der Schnittfl?che dar (Jarvis & Duncan, 1975). Die Knollen von Red Craigs Royal waren zum ersten Erntetermin reif und zeigten danach wenig ?nderungen in der Wundverheilungsrate. Die Knollen von Record hatten unreif eine langsame Wundverheilungs-Rate, bei sp?teren Ernteterminen wurde das Wundverheilen progressiv schneller, haupts?chlich bei Knollen aus den abget?teten Parzellen, und erreichte zum l. Oktober ein Maximum (Abb. 1). Abbildung l basiert auf über 10 Tage Wundheilung summierter interner Widerstandsf?higkeit, aber die Unterschiede zwischen den Behandlungen über die Wundheilperiode hinweg waren durchweg einheitlich (Abb. 2). Nach 5–7 Monaten Lagerung bei 10°C entwickelte sich interne Resistenz schneller als nach der Ernte, und blieb bei Record schneller als bei Red Craigs Royal. Die Unterschiede zwischen den Erntezeitpunkten waren jedoch nicht mehr eindeutig (Tabelle 1). Folglich ist die langsame Wundheilung bei unreif geernteten Knollen nur ein Kurzzeiteffekt. Beim Gewicht der Scheiben w?hrend der Wundheilung ergaben sich keine betr?chtlichen ?nderungen (Tabelle 2). Dies zeigt dass sich der Wassergehalt, welcher die Wundheilrate beeinflussen kann, nicht ?nderte.

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Résumé Le but de cette étude est d'étudier l'influence du niveau de maturité des tubercules sur le taux de cicatrisation des blessures immédiatement qprès récoltés à différentes dates, entre le 22 ao?t et le serve également après conservation. Des tubercules de la variété demi-précoce Red Craigs Toyal et de la variété principale Record ont été récoltés à différentes dates, entre le 22 ao?t et le 14 octobre 1979. Sur la moitié des parcelles, un défanage mécanique a été réalisé le 22 ao?t et le reste a été maintenu jusqu'à maturité. La senescence naturelle était pratiquement atteinte au 22 ao?t pour la variété Red Craigs Royal et au ler octobre pour la variété Record. Des lamelles de 11 mm × 4 mm dans le tissu médullaire des tubercules sont réalisées un jour après prélèvement et l'évolution de la résistance interne à la perte en eau, comme mesure de la subérisation à la surface des la melles, a été observée pendant 10 jours (Jarvis & Duncan, 1979). Les tubercules de la variété Red Craigs Royal étaient m?rs à la première date de récolte, et par conséquent, peu de différence dans le taux de cicatrisation a été observé. Les tubercules de la variété Record ont un faible taux de cicatrisation lorsqu'ils sont immatures, mais aux dernières dates d'arrachage la cicatrisation s'accélère, notamment pour les tubercules des parcelles défanées, quiatteint un taux maximal de cicatrisation vers le ler octobre (fig. 1). La figure l donne le total de la résistance interne pour 10 jours de cicatrisation, mais la différence entre les traitements est demeurée nette tout au long de cette période (fig. 2). Après une conservation de 5 à 7 mois à 10°C, la résistance interne à la perte d'eau s'effectue, plus rapidement qu'après la récolte et elle demeure plus forte avec la variété Record qu'avec Red Craigs Royal, mais la différence observée entre les dates d'arrachage ne para?t plus évidente (tableau 1). Donc, la lente cicatrisation des blessures pour les tubercules non m?rs n'est qu'un effet à court terme. Aucune modification substancielle n'est apparue dans le poids des lamelles pendant la période de cicatrisation (tableau 2), ceci montre que la teneur en eau qui peut influencer le taux de cicatrisation n'a pas évolué.

     

Polyphenolic compounds interfere with quantification of protein in soil extracts using the Bradford method

 The Bradford protein quantification assay is based on an absorbance shift in Coomassie brilliant blue G-250 (CBB). Samples extracted for glomalin, a protein produced by arbuscular mycorrhizal (AM) fungi, are quantified using the Bradford assay. CBB is known to react with polyphenolic substances, and co-extraction of glomalin and humic substances is known to occur. The effects of increasing concentrations polyphenolic compounds were measured. The addition of any amount of polyphenolic compounds increased the Bradford reactive fraction (BRF) of soil extract. Caution is required when interpreting BRF data, as comparison of BRF data from different studies or different field sites is problematic. The BRF may represent recalcitrant organic material in soil, though its relationship to AM fungi remains unclear.     

The effect of different combinations of lignocaine,ketoprofen, xylazine and tolazoline on the acute cortisol response to dehorning in calves

AIMS: The aims of this study were (a) to evaluate the effect of xylazine and tolazoline, with and without lignocaine, on the cortisol response of calves following amputation dehorning and (b) to assess the effect of a non-steroidal anti-inflammatory drug (ketoprofen) and local anaesthesia on the cortisol response of calves to amputation dehorning.

METHODS: Plasma cortisol concentrations were measured in 100 dehorned or non-dehorned 3-month-old calves over an 8-h period following five different sedative/analgesic or control treatments. Sedative/analgesic treatments were: control (no anaesthesia); local anaesthesia and ketoprofen; local anaesthesia and xylazine; local anaesthesia, xylazine and tolazoline; and xylazine only. Within each sedative/analgesic treatment group, half the calves (n=10 per group) were amputation dehorned and half were not dehorned.

RESULTS: The change in plasma cortisol concentrations in calves dehorned after being given ketoprofen and local anaesthesia did not differ significantly from that of non-dehorned control calves for at least 8 h. In contrast, the cortisol response of dehorned calves not given analgesic drugs peaked 30 min after dehorning and lasted >4 h. Xylazine injected before dehorning significantly reduced but did not eliminate the peak of the cortisol response. When both xylazine and local anaesthesia were administered before dehorning the peak in the cortisol response was virtually eliminated. In the dehorned calves that received xylazine with or without local anaesthesia, cortisol concentration increased significantly 3 h after dehorning and did not return to baseline until at least 5 h later. When tolazoline was administered shortly after xylazine, it caused a marked cortisol response, higher than the response to any other treatment.

CONCLUSIONS: Combining ketoprofen and local anaesthesia minimised the cortisol response, and by inference the pain- induced distress, following amputation dehorning in calves. Xylazine reduced the initial cortisol response to dehorning but not as much as when local anaesthesia was also given. The increase in cortisol concentration from 3–8 h after dehorning in calves given xylazine alone or in combination with local anaesthesia suggests that calves experienced pain-induced distress during this time and that xylazine had no long-term analgesic effect. Tolazoline, used to reverse the sedative effects of xylazine, caused a marked cortisol response in calves via a mechanism which remains unclear.     

Sucking behaviour of hand-reared newborn dairy calves

AIM: To determine the feeding ability of calves during the first 4 days after birth.

METHODS: The sucking behaviour of 171 dairy calves, fed from individual bottles during the first 4 days after birth, was evaluated by quantifying the volume of colostrum consumed, the duration of feeding, and speed of drinking. Calves had been separated from their mothers and brought into a rearing shed in the morning, when <24 h of age, and were offered 2 L colostrum from a bottle in the afternoon of the same day, and twice daily thereafter.

RESULTS: Newborn calves became efficient suckers from bottles within 24 h of removal from their dams (collection). On the day after birth, 95% of calves drank the 2 L of colostrum offered during the afternoon feed, and by Day 4 virtually all calves (99%) drank this amount. Calves that had inadequate colostrum from their dam were more likely to drink all 2 L offered after collection, but a small number of calves that had not had colostrum from their dams and drank <500 ml at the afternoon feed following collection were likely not to drink 2 L on the following days. However, calves that did not drink all 2 L on Day 1 were not disadvantaged in comparison to those that did, in that they were equally likely to drink 2 L on Day 4.

CONCLUSIONS: The majority of calves became efficient drinkers within 48 hours of birth, but a small number of slow feeders may need individual feeding at least up to 4 days after birth.