Identification of appropriate reference genes for qPCR analyses of porcine placentae and endometria,supplying foetuses of different size and sex,at multiple gestational days

Recent studies suggest associations exist between foetal size and sex, and gene expression at the porcine feto-maternal interface. It is essential to identify reference genes which have stable expression throughout gestation in feto-placental units associated with foetuses of different size and sex. qPCR was performed for 11 genes within porcine placentae and endometria at gestational days (GD) 30, 60 and 90. Several reference genes were found to have stable expression in these samples. The combination of B2m1 and Tbp1, and Hprt1 and Tbp1 had the most stable expression in endometria and placentae, respectively. Reference genes identified as having stable expression were utilized in a larger experiment with placentae and endometria associated with foetuses of different size and sex at four GD. The average expression of B2m1 and Tbp1 mRNAs was suitable for the normalization of temporal changes in endometria, and comparison between endometria supplying foetuses of different size throughout gestation. The average expression of Hprt1 and Tbp1 mRNAs was suitable for the normalization of placental mRNA expression for comparison of temporal changes and sex differences between placentae supplying foetuses of different sex throughout gestation. This combination was suitable for the normalization of mRNA expression in placentas supplying GD30, GD60 and GD90 foetuses of different size. This study has identified reference genes with stable expression in placentae and endometria across multiple gestational days, in tissues associated with foetuses of different size and sex. The results of these experiments highlight the importance of selecting appropriate reference genes for the biological comparison under investigation.     

Validation study of gas chromatographic determination of pentachlorophenol in animal liver

A validation study was conducted of a gas chromatographic procedure for the determination of pentachlorophenol (PCP) in chicken, pork, and beef liver. Five analysts representing 5 laboratories analyzed randomly numbered blind duplicates at 3 fortified tissue concentrations and one incurred tissue on 2 consecutive days. The PCP concentrations ranged from approximately 40 to 400 parts per billion (ppb). All data were reported to 3 significant figures in ppb. The coefficients of variation for repeatability were between 2.8 and 8.5%, except for the beef liver, at a mean value of 80 ppb PCP, where the CV was 11.3%. The CVs for reproducibility were in the range of 9.7-16.5% with little significant difference by species. The CV asymptotically approached 10% as the PCP level increased.     

A comparison of the pathogenicity of two strains of hog cholera virus: 1. Clinical and pathological studies

The virulence of a strain of hog cholera virus isolated during an outbreak of mild disease in pigs in New South Wales in 1960/61 (the NSW strain) was compared over 11 days with that of a virulent strain by inoculating 8 pigs with each virus and comparing the ensuing clinical signs and pathology. Both viruses caused persistent pyrexia and leukopenia, the NSW strain 4 to 5 days and the virulent strain 3 days, after inoculation. Few other clinical signs were observed in the pigs inoculated with the NSW strain. In contrast, all pigs inoculated with the virulent strain became progressively depressed and incoordinated, and were killed between days 6 and 9. Bronchopneumonia and swollen, reddened lymph nodes were observed in pigs inoculated with both viruses. Few other gross lesions were observed with the NSW strain, but some pigs receiving the virulent strain had pustules in the tonsil and the anterior oesophagus, petechial haemorrhages in the kidney, and small infarcts at the margins of the spleen. There were marked differences in the histopathology, both in the variety of organs affected and the severity of lesions in individual organs. Suppurative bronchopneumonia occurred in both groups. Other changes in the pigs affected with the NSW strain were colitis, mild cerebral vasculitis, necrosis, haemorrhage and neutrophil infiltration in some lymph nodes and spleens. In pigs infected with virulent virus the cerebral vasculitis was so severe that there was necrosis of cells within the vessel walls.(ABSTRACT TRUNCATED AT 250 WORDS)     

A comparison of the pathogenicity of two strains of hog cholera virus: 2. Virological studies

Quantitative and qualitative differences were demonstrated in the amount of virus in a range of tissues from pigs infected with either the Weybridge or New South Wales (NSW) strains of hog cholera (HC) virus. The titre of the Weybridge strain in samples, as assessed by either virus titration in cell culture or by the density of specific fluorescing cells in tissue sections, was higher than that for the NSW strain. This correlated with the greater severity of the clinico-pathological syndrome induced by the Weybridge strain. The implications of the differences in the virus content of tissues in the diagnosis of HC is discussed as is the use of monoclonal antibodies to differentiate HC and bovine virus diarrhoea viruses.     

Paleolimnology of late quaternary deposits: seibold site, north dakota

A unique late Quaternary lacustrine deposit has been discovered recently on the Missouri Coteau of North Dakota. A diverse, extremely well-preserved biota of more than 160 species has been recovered primarily from an organic mud deposited about 9500 years before the present. The lacustrine body shallowed gradually as the climate became drier.     

Lythrum hyssopifolia (lesser loosestrife) poisoning of sheep in Victoria

Objective To document an ovine disease attributed to the consumption of Lythrum hyssopifolia (lesser loosestrife). Procedures Historical and histological review of field and experimental cases. Results 1–20% mortality occurred in sheep flocks grazing paddocks where L. hyssopifolia was the predominant green vegetation. Well‐documented disease outbreaks occurred in summer on nine farms across Victoria between 1974 and 2002. Liver damage occurred in all nine outbreaks, with kidney damage in at least eight. Hepatocyte necrosis was usually zonal to midzonal (zone 2) in the liver samples from four farms and periacinar (zone 3) in those from three farms, but some livers showed only single‐cell necrosis. Multinucleate hepatocytes near necrotic areas were a feature in six cases. Proximal tubular epithelium appeared to be the primary renal target and brown granules were often present in renal tubules. Biochemical and histological evidence of liver and kidney damage was obtained from two sheep experimentally pen‐fed harvested L. hyssopifolia. Conclusion Chemicals in L. hyssopifolia are toxic to ovine hepatocytes and renal tubular epithelial cells.     

Model performance of tree height-diameter relationships in the central Congo Basin

Key message

Tree heights in the central Congo Basin are overestimated using best-available height-diameter models. These errors are propagated into the estimation of aboveground biomass and canopy height, causing significant bias when used for calibration of remote sensing products in this region.

Context

Tree height-diameter models are important components of estimating aboveground biomass (AGB) and calibrating remote sensing products in tropical forests.

Aims

For a data-poor area of the central Congo Basin, we quantified height-diameter model performance of local, regional and pan-tropical models for their use in estimating AGB and canopy height.

Methods

At three old-growth forest sites, we assessed the bias introduced in height estimation by regional and pan-tropical height-diameter models. We developed an optimal local model with site-level randomizations accounted for by using a mixed-effects modeling approach. We quantified the error propagation of modeled heights for estimating AGB and canopy height.

Results

Regional and pan-tropical height-diameter models produced a significant overestimation in tree height, propagating into significant overestimations of AGB and Lorey’s height. The pan-tropical model accounting for climatic drivers performed better than the regional models. We present a local height-diameter model which produced nonsignificant errors for AGB and canopy height estimations at our study area.

Conclusion

The application of general models at our study area introduced bias in tree height estimations and the derived stand-level variables. Improved delimitation of regions in tropical Africa with similar forest structure is needed to produce models fit for calibrating remote sensing products.

     

The first case of genetically confirmed monozygotic twinning in the dog

Monozygotic twinning has not previously been genetically confirmed in the dog. This case report describes the finding of two viable male monozygotic foetuses within one placental site during caesarean section. Their umbilical cords attached to a single placenta. Genetic profiling using a total of 38 microsatellite markers, as well as amelogenin and SRY for sex determination, revealed identical DNA profiles, whether derived from blood or tissue (buccal swabs) samples. To the best of our knowledge, this is the first report of monozygotic twinning in the dog confirmed using DNA profiling.     

Recent developments in equine dentistry

ABSTRACT

Equine dentistry is a rapidly developing clinical specialty. It has benefitted from key advances in anatomical and physiological research, development of equipment and instrumentation, utilisation of standing sedation and anaesthesia protocols, a change towards minimally invasive surgical techniques, and the introduction of restorative and endodontic techniques translated from techniques used in human and canine dentistry. Anatomical research has provided further insight into the endodontic system of incisors and cheek teeth and how it changes throughout development with age. Studies of the periodontium have demonstrated a rich vascular supply and repair capacity. Routine dental visits are increasingly being performed utilising sedation and clinical instruments for routine examinations. Equipment has become more efficient, battery-operated and miniaturised giving benefits to both equine dental maintenance work and advanced techniques, assisting the transition to minimally invasive techniques, and the development of endodontic and restorative dentistry. Diagnosis has also benefitted from advances in equipment such as patient-side digital radiography systems, high definition oroscopy, and small diameter flexible fibrescopes that are capable of visualising inside a pulp canal. Dental units combining endodontic high- and low-speed drills, suction and air or water flush are becoming increasingly used and adapted for equine use. Sedative combinations and standing anaesthesia protocols have meant that revisions of traditional techniques, as well as novel techniques, can be performed with almost no requirement for general anaesthesia. Equine dentistry can only continue to advance in this way if there is early identification of dental disease through clinical oral examinations, leading to a system based on prophylaxis, as in human dentistry. This necessitates a change in attitude of the public and industry in general to a proactive approach, with early intervention based on examination and diagnostic findings of practitioners and not necessarily based on the clinical signs displayed by the patient.     

Expression of Occludin in Testis and Epididymis of Wild Rabbits, Lepus sinensis coreanus

Tight junctions (TJs) in inter-Sertoli junctional areas and epididymal epithelia build up the blood–testis barrier (BTB) and the blood–epididymal barrier (BEB), respectively. In this study, the expression of occludin, an integral member of the TJs, was examined in testis and different regions of epididymis of Lepus sinensis coreanus , an Korean wild rabbit species. In testis, intense occludin immunoreactivity was found in the basally located inter-Sertoli junctional area together with diffused immunoreactivity of occludin in the cytoplasm of Sertoli cells. It can be suggested that occludin is one of the robust elements of BTB in seminiferous tubules of rabbit testis. In proximal and distal caput epididymis, occludin immunoreactivity was found in the lateral as well as apical contacts of epithelial cells. In corpus epididymis, intense occludin immunoreactivity was found in the basolateral as well as apical contacts of epithelial cells together with cytoplasmic signal. In cauda epididymis, occludin immunoreactivity in luminal epithelia was relatively strong but largely found in the cytoplasm. This suggests that intriguing regulatory mechanisms differentially recruit occludin to the TJ in the different regions of epididymal epithelia. The differences in the subcellular localization as well as expression levels of occludin among the epididymal segments may reflect differential paracellular permeability of epithelia along the epididymal tubules and be correlated with sperm maturation in rabbit. In Western blot, a major form of occludin was MW 62 kDa together with small fragments of MW 34–39 kDa in testis and epididymis, suggesting the peptide cleavage of occludin. This is the first report on the molecular nature of TJs in a wild rabbit testis and epididymis.