Exogenous DNA Uptake of Boar Spermatozoa by a Magnetic Nanoparticle Vector System

The sperm‐mediated gene transfer method is applicable to transgenesis in many species that use spermatozoa for reproduction recently, which has been shown various results. In the current study, we show that transgenic porcine embryos can be efficiently produced by employing a simple transfection method that uses magnetic nanoparticles (MNPs). The complexes formed between plasmid DNA and MNPs were bounded on ejaculated boar spermatozoa at a higher efficiency compared to methods using DNA alone or lipofection. Using confocal microscopy, rhodamine fluorophore‐labelled MNPs were detected on external surfaces of the spermatozoa membrane, which were bounded on zona pellucida of in vitro maturated oocyte during in vitro fertilization. Electron microscopy revealed that clusters of MNPs were detected in inside of plasma membrane and nucleus of the spermatozoa head. Additionally, we found that magnetofected boar spermatozoa could be fertilized with oocytes in vitro and that the resulting gene of green fluorescent protein was detected in fertilized eggs by genomic PCR analysis. Taken together, these results suggest that MNPs can be used to efficiently introduce a transgene into embryo via spermatozoa.     

Nasopharyngeal cryptococcosis

Naturally occurring cryptococcosis in five cats, a dog and a koala is described. Involvement of the nasopharynx was documented in all patients, and nasopharyngeal mass lesions accounted for the major presenting complaints in four. Signs referable to nasopharyngeal disease included snoring, stertor, inspiratory dyspnoea and aerophagia. Diagnoses were made by caudal rhinoscopy using a retroverted flexible endoscope, vigorous orthograde flushing with saline, or at necropsy. Concurrent cryptococcal rhinitis was present in all cases, although involvement appeared limited to the caudal nasal cavity in most cases. Typical signs of nasal cavity disease, such as sneezing and nasal discharge, were often absent. Treatment of nasopharyngeal cryptococcosis should include physical dislodgement or debulking of lesion(s) to provide immediate alleviation of upper airway obstruction, followed by systemic antifungal therapy to eliminate residual infection from the nasal cavity. Infections caused by Cryptococcus neoformans var gattii accounted for a disproportionately large number of these cases.     

International approach to plant exploration and introduction

These is a need to review the theories of N. I. Vavilov on the basis of modern knowledge on the nature and genetics of variability, ecogenetics, environmental archaeology and related subjects.Greater care should be taken in the preliminary planning of plant collecting expeditions, so that collectors may be better informed on the ecology of the species they are to collect, and of the types of vegetation in which they are likely to occur. The best method of sampling populations should be evolved so that plant introduction stations shall not be swamped with vast collections.Current interest is widespread with regard both to species and to the countries concerned. There is a great need for new material of herbage and fodder plants, grain legumes, cereals, potato, coffee, cacao, rubber and other crops. Time is short, as the centres of origin are suffering from clearance of vegetation and development.There is a recent tendency to propose the establishment of Plant Exploration Centres within the centres of origin.

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Samenvatting Er bestaat aanleiding de theorieën van N. I. Vavilov te herzien op grond van nieuwe kennis omtrent de aard en de genetische achtergrond van de variabiliteit, de ecogenetica, de archeologie van het milieu en dergelijke.Meer zorg dient te worden besteed aan de voorbereiding van expedities opdat de plantenzoekers beter ingelicht zijn over de ecologie van de soorten, die ze gaan verzamelen, en over de vegetatie-typen waarin ze waarschijnlijk voorkomen. De beste methoden dienen te worden ontwikkeld om monsters uit populaties te verzamelen opdat de planten-introductie-centra niet overstelpt worden met materiaal.De belangstelling richt zich overal op de soorten zowel als op de landen. Er is grote behoefte aan nieuw materiaal van voedergewassen, peulvruchten, granen, aardappels, koffie, cacao, rubber en andere gewassen.De tijd dringt, aangezien de genencentra bedreigd worden met uitputting door het opruimen van vegetaties en door ontwikkeling.Er is thans een tendens merkbaar om voorstellen te doen tot het vestigen van verzamelcentra in de oorsprongsgebieden zelf.

     

Diet selection by sheep and goats and sward composition changes in a ryegrass/white clover sward previously grazed by cattle, sheep or goats

An experiment was conducted to examine how variation in the composition and structure of mixed grass/white clover swards affected diet selection by sheep and goats. Sward composition in a mixed perennial ryegrass/white clover sward was manipulated by continuous grazing from 28 May to 28 July (pre-experimental phase) with cattle, sheep or goats, and then from 29 July to 2 September (experimental phase) with sheep or goats in a factorial design replicated twice. Sward surface height was maintained at 6 cm by regular adjustment of stocking density. Grazing by different sequences of animal species resulted in significant differences in the proportions of white clover in the sward, and especially in the proportion of clover lamina and petiole. Grazing by goats in the pre-experimental phase led to greater proportions of clover lamina and petiole in the whole sward and the sward surface. The proportion of white clover in the diet selected by sheep in the experimental phase was consistently higher than that in the sward as a whole, but was closely related to that near to the sward surface (approximately the top 2 cm). For goats there was no significant relationship between the proportion of clover in the diet and in the whole sward, and they generally selected a diet with a lower proportion of white clover than was present in approximately the top 2 cm of the sward. It is concluded that on mixed grass/white clover swards goats do not graze as deeply into the sward as sheep and that this results in a lower proportion of white clover in their diet and therefore allows higher proportions of white clover to develop under grazing by goats than by sheep.     

Plasma cortisol responses to three cortico-trophic preparations in normal dogs

Objective To compare cortisol responses to three corticotrophic preparations in normal dogs.
Animals Eight clinically normal dogs (four intact males, four intact females) of medium size.
Procedures Each dog received four treatments on four separate occasions in a duplicated Latin square pattern. Treatments were two adrenocorticotrophin (ACTH) preparations given intramuscularly at 2.2 U/kg, one of the ACTH preparations given intramuscularly at 1 U/kg and a synthetic polypeptide with ACTH-like activity (tetracosactrin, cosyntropin) given intravenously at 5 μg/kg. Plasma samples were taken for cortisol assay before and at 0.5, 1, 2 and 4 h after treatment.
Results Plasma cortisol concentrations were similar with the two ACTH preparations and at both dose rates. Tetracosactrin produced smaller mean peak cortisol concentrations, which tended to occur earlier than with ACTH, and smaller values for the area under the curve of plasma cortisol concentration from zero time to 4 h.
Conclusion The findings suggest that canine adrenal function can be tested adequately by giving ACTH intramuscularly at 1 U/kg and measuring plasma cortisol in samples taken at 0 and 2 h, or by giving tetracosactrin intravenously at 5 μg/kg and determining cortisol concentration at 0 and 1 h.     

Effects of delmadinone acetate on pituitary-adrenal function, glucose tolerance and growth hormone in male dogs

Objective To characterise the effects of delmadinone acetate on the pituitary-adrenal axis, glucose tolerance and growth hormone concentration in normal male dogs and dogs with benign prostatic hyperplasia.
Design A prospective study involving nine normal male dogs and seven with prostatic hyperplasia.
Procedure Delmadinone acetate was administered to six normal male dogs and seven dogs with benign prostatic hyperplasia at recommended dose rates (1.5 mg/kg subcuta-neously at 0, 1 and 4 weeks). Three normal controls received saline at the same intervals. Blood concentrations of ACTH, cortisol, glucose, insulin and growth hormone were measured over 50 days. Intravenous glucose tolerance and ACTH response tests were performed before and after treatment in the nine normal animals.
Results A substantial suppression of basal and 2 h post-ACTH plasma cortisol secretion was demonstrated after one dose in all dogs given delmadinone acetate. Individual responses after the second and third administration varied between recovery in adrenal responsiveness to continued suppression. Plasma ACTH concentration was also diminished after one treatment. No effects were evident on glucose tolerance or serum growth hormone concentrations.
Conclusion Delmadinone acetate causes adrenal suppression from inhibition of release of ACTH from the pituitary gland. Treated dogs may be at risk of developing signs of glucocorticoid insufficiency if subjected to stressful events during or after therapy. Neither glucose intolerance nor hyper-somatotropism seems likely in male dogs given delmadinone acetate at the recommended dose rate, but the potential for excessive growth hormone secretion in treated bitches remains undetermined.     

Reliably colonising broiler chickens with Campylobacter spp. using a litter-based method

ABSTRACT

1. Chicken-associated Campylobacter spp. are the cause of most food poisoning cases in Europe. In order to study the host–pathogen interactions, a reliable and reproducible method of colonising chickens with the bacteria is required.

2. This study aimed to identify a more appropriate and less invasive method of colonisation (cf. gavaging) by seeding bedding material (litter) that commercial chickens are kept on with a mixture of Campylobacter spp., broth and faeces.

3. The first phase of the study tested the longevity of Campylobacter spp. recovery in seeded litter over 24 h: significantly more Campylobacter spp. was recovered at 0 or 3 h post-seeding than at 6 and 24 h post-seeding, indicating that the pathogen can survive to detectable levels for at least 3 h in this environment.

4. In the second phase, three groups of 10 broiler chickens (negative for Campylobacter spp. prior to exposure) were exposed at 21 days of age to one of three different Campylobacter jejuni and C. coli mixes (A, B, C), using the method above. At 28 days of age, birds were euthanised by overdose of barbiturate or cervical dislocation, and livers and caeca removed for Campylobacter spp. assessment.

5. All liver and 28/30 caeca samples tested positive for Campylobacter spp., with mix A and C giving higher counts in the caeca than mix B. The method of euthanasia did not affect Campylobacter spp. counts.

6. In conclusion, a successful method for reliably colonising broiler chickens with Campylobacter spp. has been developed which negates the need for gavaging and is more representative of how contamination occurs in the field.