Comparison of methods for differentiation of Salmonella enterica serovar Enteritidis phage type 4 isolates

OBJECTIVE: To compare molecular typing methods for the differentiation of Salmonella enterica serovar Enteritidis phage type (PT) 4 isolates that allowed for the determination of their genetic relatedness. SAMPLE POPULATION: 27 Salmonella Enteritidis PT 4 strains isolated in the United States and Europe. PROCEDURE: Several molecular typing methods were performed to assess their ability to genetically differentiate among Salmonella Enteritidis PT 4 isolates. Results of pulse-field gel electrophoresis (PFGE), repetitive polymerase chain reaction (PCR) assay, 16S rRNA gene sequencing, random amplification of polymorphic DNA (RAPD), PCR-restriction fragment length polymorphism of 16S rRNA, and antimicrobial susceptibility were evaluated. RESULTS: Compared with results for other techniques, results for the RAPD typing method with the RAPD1 primer reveal that it was the most discriminatory fingerprinting technique, and it allowed us to cluster Salmonella Enteritidis PT 4 isolates on the basis of their genetic similarity. CONCLUSIONS AND CLINICAL RELEVANCE: This study revealed the value of RAPD with the RAPD1 primer as a tool for epidemiologic investigations of Salmonella Enteritidis PT 4. It can be used in conjunction with PFGE and phage typing to determine the genetic relatedness of Salmonella Enteritidis isolates involved in outbreaks of disease. A reliable and highly discriminatory method for epidemiologic investigations is critical to allow investigators to identify the source of infections and consequently prevent the spread of Salmonella Enteritidis PT 4.     

Comparison of the Johne''s absorbed EIA and the complement-fixation test for the diagnosis of Johne''s disease in cattle

A commercially available absorbed ELISA for the diagnosis of Johne's disease (JD) (paratuberculosis) in cattle, the Johne's Absorbed EIA, was compared with the conventional complement-fixation test (CFT) used in Australia. Stored plasma from 3 Victorian dairy herds with a history of JD, sera from specimens submitted from animals showing clinical signs of JD and sera from the US National Repository for Paratuberculosis Specimens were used to determine the sensitivity of each test. The EIA detected 48.8% of 43 Australian animals with subclinical JD, while the CFT detected only 12 (21.4%) of 56 subclinically affected cattle. Of 150 subclinically infected US cattle, the EIA detected 47.3% and the CFT detected 52.0%. The EIA detected 59.7% of animals which at the time of sampling were shedding Mycobacterium paratuberculosis in their faeces, but showed no clinical signs of JD, while the CFT detected 57.3%. The EIA correctly identified 88.2% of 136 histologically confirmed clinical cases, and the CFT detected 83.4%. The specificity of each test was determined by testing sera collected at slaughter from animals residing in a known JD-free area of Australia, and from samples from the US National Repository of Paratuberculosis Specimens collected from certified-free herds in Wisconsin. The EIA was found to have a specificity of 99.8% when 998 Australian animals were used as the test population, and 99.0% when 196 US animals were used. The specificity of the CFT using Australian samples was 96.9% and 95.2% using American samples.     

Distribution of intramuscularly administered erythromycin into subcutaneous tissue chambers before and after inoculation with Pasteurella haemolytica

Distribution of erythromycin into subcutaneous tissue chambers was characterised pharmacokinetically and the effect of Pasteurella haemolytica infection on the extent of penetration was studied. Thermoplastic tissue chambers were implanted subcutaneously in the paralumbar fossae of six calves. Thirty-five days after implantation, the tissue chamber distribution of intramuscularly administered erythromycin (30 mg kg⁻¹) was studied. Chambers were then inoculated with P haemolytica and the tissue chamber pharmacokinetics of erythromycin were again studied. Diffusion of erythromycin into tissue chambers was best described using a two-compartment model with tissue chambers representing a relatively inaccessible compartment. Despite changes in chamber fluid pH, the extent of erythromycin penetration into chambers was not affected by P haemolytica inoculation. Comparison of computer simulated concentration-time curves resulting from different routes of administration revealed that penetration of erythromycin into less accessible sites was more likely to be higher after intravenous administration than after intramuscular administration.     

Attitudes of veterinarians to undergraduate education in genetics and the use of genetics in veterinary occupations

A questionnaire regarding attitudes to undergraduate education in genetics and the subsequent use of that education was sent to 1000 veterinarians registered in New South Wales. Three hundred replies to the questionnaire were received and analysed. No significant difference in the perceived adequacy of courses at the four universities involved in teaching veterinary science was observed. Opinions on adequacy of education in genetics were not affected by age, sex or years since graduation. The major reason cited for lack of adequacy was that undergraduate genetics courses were not practical. Only 17 respondents stated that they were never approached for information on genetics or animal breeding, while 76 were approached more than 20 times annually for information. Eighty-three respondents claimed to deal with more than 20 cases annually that required some knowledge of genetics. The results of the questionnaire highlight the importance of genetics instruction in undergraduate veterinary education. The questionnaire responses also provide insight into how working veterinarians consider that courses in genetics could be improved.     

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.     

Genomic Imprinting in Canis familiaris

For the vast majority of mammalian genes, maternally- and paternally-derived alleles behave identically and are either expressed or repressed, regardless of whether they were inherited from egg or sperm. For imprinted genes, however, this is not the case. The alleles of imprinted genes are epigenetically modified in a parent-of-origin-specific manner and, as a consequence, maternally- and paternally-derived alleles behave differently. Typically one allele is expressed while the other is silent. Although relatively few in number, imprinted genes are the focus of intensive study, as they have important roles in embryonic development. Abnormal expression of imprinted genes results in growth disorders and is implicated in several clinical conditions. Most studies of imprinted genes have been performed in rodents or primates, with limited studies in other mammals such as bovine and opossum. We have recently demonstrated the existence of imprinted genes in the canine, by showing that the canine insulin-like growth factor 2 receptor gene ( IGF2R ) is monoallelically expressed, with predominant expression of the maternally-derived allele and repression of the paternally-inherited allele. Our ultimate goal is to characterize all imprinted genes in the canine, and to understand how they contribute to canine reproduction, development and disease. Such knowledge will be vital for optimizing the success of most reproductive strategies in the canine.     

Post-Partum Involution of the Canine Uterus – Gross Anatomical and Histological Features

We aimed to study the normal puerperium in the bitch. Ovariohysterectomy was performed in nine bitches, each at a different day after normal whelping; their genital tract was subject to gross anatomical examination, as well as to histological examination and electron microscopy scanning. Corpora albicans were evenly distributed in the left and right ovaries and placental sites were evenly distributed among left and right uterine horns. Placental sites were initially of dark green to grey colour, later becoming dark brown; their length and height progressively decreased. Height of the myometrium and diameter of the uterine glands progressively decreased. Trophoblast-like cells were consistently observed at the placental sites and on the surface of the interplacental areas, at all time points where hysterectomy had been performed. It is suggested that involution of the canine genital tract can last up to 3 months and is slow. Continuous (up to D84 post-partum) presence of prominent placental sites should be considered a normal feature of canine uterine post-partum involution.     

Breeding behavior of yield components and hollow heart in tetraploid-diploid vs. conventionally derived potato hybrids

Summary A factorial mating design with six 4x cultivars as stylar parents mated to three groups of pollen parents (4x cultivars, 4x high protein selections, and 2x S. phureja) was used to study breeding behavior of yield, tuber set, marketable yield and hollow heart in potatoes. Mean yield for the 4x-2x hybrids was equivalent to that of their cultivar parents although yield components, tuber set and average tuber size, were greater and less, respectively, than the cultivar parents. High estimates of general combining ability were found in both pollen and stylar parents for yield, tuber set, marketable yield, number of marketable tubers and hollow heart in all three hybrid populations. Specific combining ability estimates were significant only for yield and hollow heart. Frequency of white-fleshed 4x-2x progeny of five heterozygous yellow-fleshed 2x phureja genotypes was used to confirm cytological identification of a first division restitution mechanism operative in the 2x parents. Although three of these phureja genotypes generated high-yielding progeny, two others produced 4x-2x progeny with mean yield similar to that of hybrids among 4x cultivars.Scientific journal series article 11,601 of the Minnesota Agricultural Experiment Station.