Rapid estimation of the amylose/amylopectin ratio in small amounts of tuber and leaf tissue of the potato

Summary Small amounts of potato tuber and leaf tissues are extracted in perchloric acid. After staining with I₂-KI solution absorbancies at 618 and 550 nm are measured. The amylose/amylopectin ratio can be estimated from the ratio of the absorbancies by using a formula or a graph in which the specific absorptions of the two compounds are introduced. Weighing of samples is not required. The method can be used for the estimation of the starch composition of minitubers and is especially suitable for the detection of mutants with an altered starch composition.     

Spirometra erinacei / S. erinaceieuropaei in a feral cat in Manawatu with chronic intermittent diarrhoea

CASE HISTORY: A feral cat captured in the Manawatu region of New Zealand was treated for worms and fleas, and kept confined in a metabolic cage. It showed good appetite and weight gain but had intermittent watery, yellow diarrhoea.

CLINICAL FINDINGS: Clinical examination under sedation was unremarkable and routine blood tests showed no significant abnormalities. The cat was negative for feline immunodeficiency virus (FIV) and feline leukaemia virus (FeLV). Different canned cat foods did not alter the course of the diarrhoea, and the cat was euthanised 6 months after capture. At necropsy, two sections of adult Spirometra tapeworms were found in the jejunum and typical Spirometra eggs were found in colonic contents. Molecular identification of the parasite was undertaken, using the cytochrome- c oxidase subunit-1 gene (cox1) sequence.

DIAGNOSIS: Chronic intermittent diarrhoea associated with Spirometra erinacei / S. erinaceieuropaei infection.

CLINICAL RELEVANCE: Spirometra has not been reported in New Zealand before but has been associated with gastrointestinal disease in cats in other parts of the world. It requires speciestargeted treatment to be eliminated effectively, and is zoonotic. Diagnosis could be diffi cult for clinicians who are not familiar with the parasite and its life cycle.     

Distal aortic aneurysm presumed to be secondary to an infected umbilical artery in a foal

Abstract

CASE HISTORY:?A 3-month-old female Warmblood foal was presented after displaying signs of colic with pyrexia for 5 days.

CLINICAL AND PATHOLOGICAL FINDINGS:?The foal continued to show signs of colic, frequently passed urine, and was pyrexic with an elevated white blood cell count. The umbilical stalk was thickened but there was no evidence of purulent material. Exploratory laparotomy revealed an enlarged left umbilical artery remnant tightly adhered to the bladder wall. The left umbilical artery continued to an aneurysm involving the distal aorta. The foal was subject to euthanasia and post-mortem examination confirmed a spherical aortic aneurysm, in the dorsal midline caudal to the kidneys that contained a large thrombus. Histopathological examination revealed inflammation and necrosis of the tunica intima and tunica media of the left umbilical artery with suppuration and bacterial colonies evident in the periarterial tissues.

DIAGNOSIS:?Infected aortic aneurysm presumably caused by an umbilical artery infection.

CLINICAL RELEVANCE:?A previously undetected umbilical infection appears to have resulted in an unusual delayed complication causing signs of colic in a foal. Veterinarians should be aware of this condition, and the possibility that it may be a cause of signs of colic in foals. Diagnosis based on ultrasonography should be possible, but may require sedation, visceral analgesia and careful examination.     

Expression of Apoptosis Regulatory Genes and Incidence of Apoptosis in Different Morphological Quality Groups of In Vitro‐produced Bovine Pre‐implantation Embryos

Apoptosis occurs during early development in both in vivo‐ and in vitro‐produced embryos, and is considered as one of the causes of embryonic loss. The objectives of this study were, therefore, investigating stage‐specific expression profiles of apoptosis regulatory genes in three quality groups of in vitro‐produced bovine pre‐implantation embryos; and analysing the relationship between cell number and DNA fragmentation with expressions of those genes. The relative abundance of mRNA of 9 pro‐ (Bax, caspase‐9, Bcl‐xs, P53, Caspase‐3 and Fas) and anti‐ (Bcl‐w and Mcl‐1) apoptotic genes was analysed. Differential cell staining and terminal deoxynucleotidyl transferase‐mediated dUTP‐biotin nick end labelling were performed to analyse the variation in cell numbers and detect apoptotic nuclei respectively. Expression of Bax and Caspase‐3 genes was significantly (p < 0.05) higher in poor quality pre‐implantation embryos as compared with that of morphologically good quality embryos of the same developmental stages. Moreover, Mcl‐1 expression was significantly higher in good quality immature oocytes than that in the poor quality group. Moreover, higher DNA fragmentation was evidenced in morphologically poor quality blastocysts. In conclusion, our study demonstrates that Bax, caspase‐3 and Mcl‐1 can be used as potential markers of embryo quality to evaluate in vitro‐produced bovine embryos. Further studies are required to investigate specific molecular signatures that can be used in evaluating in vivo‐derived embryos.     

Environmental influences on daily commercial catch rates of South Australia's southern rock lobster (Jasus edwardsii)

The extent to which catchability of southern rock lobster (Jasus edwardsii) due to short‐term environmental factors, rather than abundance, may be affecting legal‐size catch rates from the South Australian Southern Zone rock lobster fishery was examined. Multivariate weighted linear regression was applied to daily aggregated commercial catch rates using several environmental covariates in addition to year and month. Model pruning via backward selection identified the following variables as being significantly related to catch rate: wave height and period, lagged wave height, bottom temperature, moon phase, and a spatial block index. These variables explained 7% of the total variance in log‐transformed daily catch rates and another 84% was explained by month and year. A negative relationship was found between catch rate and each of bottom temperature and same‐day wave height, while the relationships between catch rate and days prior to full moon, and average wave height over the past 3 days were positive.     

A three‐dimensional alginate system for in vitro culture of cumulus‐denuded feline oocytes

In the case of high valuable individuals with very precious genetic material, widening the genetic pool including gametes with poor morphological characteristics, as cumulus‐denuded oocytes (CDOs), could be an option. To improve the in vitro culture of low‐competence feline CDOs, an enriched three‐dimensional (3D) system in association with competent cumulus–oocyte complexes (COCs) was developed. For this purpose, domestic cat CDOs were cultured with or without companion COCs in the 3D barium alginate microcapsules. The overall viability and the meiotic progression of feline CDOs cocultured with COCs or cultured separately in 3D or in 2D (traditional microdrops) system were compared. The 3D system was able to support viability and meiotic resumption of the feline oocytes, as well as the 2D microdrops. In 3D microcapsules, the presence of COCs resulted in a higher viability of CDOs (91.1%, p < .05), than that obtained without COCs or in 2D microdrops (71.2% and 67.3%, respectively), but the percentages of meiotic resumption were similar of those of CDOs cultured separately (55.4% vs. 40.4%, p > .05). It is notable that the presence of CDOs seemed to enhance the meiotic progression of the associated COCs. In conclusion, the 3D barium alginate microcapsules are a suitable system for feline oocytes in vitro culture, but more specific enriched conditions should be developed to improve the CDOs full competence in vitro.     

Corpus Luteum–Endometrium–Embryo Interactions in the Dairy Cow: Underlying Mechanisms and Clinical Relevance

Conception rates of dairy cows are currently declining at an estimated 1% every year. Approximately, 35% of embryos fail to prevent luteolysis during the first three weeks of gestation. Interactions between the corpus luteum, endometrium and embryo are critical to the successful establishment of pregnancy and inadequacies will result in the mortality of the embryo. For example, as little as a one day delay in the post-ovulatory rise of progesterone has serious consequences for embryo development and survival. Recently, we found that LH support, degree of vascularization and luteal cell steroidogenic capacity were not the major factors responsible for this luteal inadequacy, but are nevertheless essential for luteal development and function. Progesterone acting on its receptor in the endometrium stimulates the production of endometrial secretions on which the free-living embryo is dependent. However, their exact composition and effects of inadequate progesterone remains to be determined. The embryo is recognized through its secretion of interferon tau (IFNT), which suppresses luteolytic pulses of prostaglandin F_2α. In the cow, it is most likely that IFNT inhibits oxytocin receptor up-regulation directly and does not require the prior inhibition of oestrogen receptor α (ESR1). Unravelling the precise luteal-endometrium and embryo interactions is essential for us to understand pregnancy establishment and development of strategies to reverse the declining fertility of dairy cows.