Fine mapping quantitative trait loci under selective phenotyping strategies based on linkage and linkage disequilibrium criteria

In fine mapping of a large‐scale experimental population where collection of phenotypes are very expensive, difficult to record or time‐demanding, selective phenotyping could be used to phenotype the most informative individuals. Linkage analyses based sampling criteria (LAC) and linkage disequilibrium‐based sampling criteria (LDC) for selecting individuals to phenotype are compared to random phenotyping in a quantitative trait loci (QTL) verification experiment using stochastic simulation. Several strategies based on LAC and LDC for selecting the most informative 30%, 40% or 50% of individuals for phenotyping to extract maximum power and precision in a QTL fine mapping experiment were developed and assessed. Linkage analyses for the mapping was performed for individuals sampled on LAC within families and combined linkage disequilibrium and linkage analyses was performed for individuals sampled across the whole population based on LDC. The results showed that selecting individuals with similar haplotypes to the paternal haplotypes (minimum recombination criterion) using LAC compared to random phenotyping gave at least the same power to detect a QTL but decreased the accuracy of the QTL position. However, in order to estimate unbiased QTL parameters based on LAC in a large half‐sib family, prior information on QTL position was required. The LDC improved the accuracy to estimate the QTL position but not significantly compared to random phenotyping with the same sample size. When applying LDC (all phenotyping levels), the estimated QTL effect were closer to the true value in comparison to LAC. The results showed that the LDC were better than the LAC to select individuals for phenotyping and contributed to detection of the QTL.     

Gene cloning of porcine adiponectin gene from adipose tissue and construction of its eukaryotic expression vector

To clone adiponectin (ADPN) gene from Shaziling porcine adipocyte and construct its eukaryotic expression vector, total RNA was extracted from subcutaneous fatty tissue. One pair of specific primers was designed by Primer 5.0 software according to the sequence of ADPN gene of porcine available in GenBank. The ADPN gene was amplified by PCR from cDNA and cloned into pMD18‐T vector to construct recombinant clonal vector pMD‐ADPN, sequenced and analysed. A recombinant expression plasmid pPICZaA‐ADPN was constructed by subcloning the cloned ADPN gene into the linearized pPICZaA vector. Then, the plasmid pPICZaA‐ADPN was expressed in Pichia pastoris (GS115) by electrotransformation. Western blot and Bradford analysis were used to determine the target protein induced by methanol. Results showed that the genome size of ADPN was 732 bp and encoded 244 amino acid, the nucleotide sequence of ADPN shared 100% identity with that of porcine available in GenBank. Western blot and Bradford analysis showed that the recombinant ADPN was expressed in GS115 correctly and has certain immune activity. The expression level of ADPN was 28.5 μg/ml. In conclusion, the recombinant ADPN could express in eukaryotic expression vector pPICZaA‐ADPN constructed in this study effectively.     

Extra-adrenal paraganglioma of the equine orbit: six cases

At The Ohio State University from 1994–2006 six of seven horses evaluated for primary orbital disease were diagnosed with extra-adrenal paraganglioma (EAPG). The horses ranged in age from 14 to 24 years, with a mean of 16.8 years. Duration of clinical signs was 1.5 years to 5 years, with a mean of 2.8 years. Clinical signs varied, but all six had non-painful exophthalmus of the right eye. Five horses had complete ocular exams reported; three of five had decreased to absent vision, two of five had pale optic nerves, and in three of five, difficulty of retropulsion of the globe was noted. Diagnostic tests performed included complete blood count, serum profile, radiography, ultrasound, computed tomography, true-cut biopsy, ocular examination, guttural pouch endoscopy, oral examination, and physical examination. Expulsive hemorrhage during orbital exenteration occurred in all horses. In five of six cases, tumor extension through the orbital foramen was apparent intra-operatively. Histopathologic appearance of all surgically removed tissues consisted of sheets of polygonal cells with abundant lightly granular cytoplasm, round nuclei with vesicular chromatin, and rare mitoses. Neoplastic cells were arranged into small groups separated by a fine fibrovascular stroma. All six cases were chromagranin positive on immunohistochemical staining. Follow-up ranged from six months to six years, with a mean of two years. Four of the five horses that recovered from surgery had no apparent tumor recurrence in 6–48 months.     

Implantable loop recorders: a viable diagnostic tool in veterinary medicine

Objective : The purpose of this study was to investigate whether implantable loop recorders could be used in the diagnosis of unexplained collapse in dogs. Methods : The medical records of six dogs presented to the University of Liverpool Small Animal Teaching Hospital between May 2003 and October 2006 for further evaluation of intermittent syncopal episodes, collapse or episodic weakness, were reviewed. All these dogs underwent standard investigations and had implantable loop recorders placed. Results : A provisional diagnosis of supraventricular tachycardia was made in one dog, and diagnoses of exclusion of arrhythmogenic right ventricular cardiomyopathy and idiopathic epilepsy was made in two dogs. One dog suffered no further syncopal episodes, a diagnosis was not reached in another dog and the final dog was lost to follow‐up. Clinical Significance : The implantable loop recorder can be used successfully for the diagnosis of unexplained collapse in dogs.     

Bone strength of laying hens kept in an alternative system, compared with hens in cages and on deep-litter

1. Observations of vigorous wing movements and measurements of bone strength were compared in two experiments with birds in three different housing systems: a semi-intensive alternative system under development, a battery cage system and a deep-litter system. 2. A significant effect of housing system on the frequency of vigorous wing movements was found. The highest frequency was seen in the deep-litter system, about half this number in the alternative system, while in the battery cages they were never observed. 3. Corresponding to this a reduction in humerus strength of 9% was found in hens from the alternative system and of 45% in hens from cages, compared with deep-litter. A reduction in tibial breaking strength was also found in caged hens, when compared to deep-litter hens. 4. Keeping hens in cages thus restricts their movements, especially wing movements, to the degree that bone strength is greatly reduced. 5. This has welfare implications, for hens with low bone breaking strength risk a possibility of breakage, especially when handled and transported. When alternative systems are designed opportunities for movement in the three dimensions should be considered.     

Bee pollen and propolis as dietary supplements for rabbit: Effect on reproductive performance of does and on immunological response of does and their offspring

To evaluate the effect of bee pollen (BP) and/or propolis (Pro) supplementation on rabbit does, 64 nulliparous NZW rabbits does were distributed among eight groups (eight animals/group). One unsupplemented group was the control; the other seven groups were supplemented, respectively, with zinc bacitracin (ZnB) at 100 mg, BP at 150 and 300 mg, Pro at 150 and 300 mg, BP+Pro at 150 and 300 mg of each three times/week, day after day continuously along eight parities. The BP300, Pro300 and BP+Pro150 groups had higher body weight of litter at birth and number of kids born alive. The BP supplementation at 150 mg increased plasma total protein and albumin than the control group. The BP or Pro at 150 mg decreased plasma T₃ than the other groups except for BP+Pro150. The ZnB group had significantly greater T₃/T₄ ratio compared to BP, Pro and BP+Pro at 150 mg. The BP+Pro150 group had less ALT than the control; BP300 and Pro 300 mg resulted in lower plasma AST than the groups Pro150 with or without BP and the control group. The plasma alkaline phosphatase of BP at 150 or 300 mg and BP+Pro150 was significantly greater than that of the Pro150 group. The BP+Pro300 group had higher WBCs than the other groups. In contrast, the lymphocytes were greater in the Pro and BP+Pro300 groups than in BP, Pro and BP+Pro at 150 mg. The groups supplemented with BP and BP+Pro at 150 and 300 mg had significantly greater SRBCs of doe rabbits and their offspring compared to the control and the ZnB group. The BP at 300 mg increased the serum albumin and α₁‐globulin than the control group. The Pro300 group had greater serum α₂‐globulin and β‐globulin than the control group. The total globulin was significantly greater for the 300 mg propolis‐supplemented groups than the control.     

A comparison of the uterine proteome of mares in oestrus and dioestrus

Proteomic analysis of mare uterine flush fluid provides a minimally invasive technique for studying protein changes associated with the oestrous cycle. The aim of this study was to identify differentially abundant proteins in the uterine flush fluid of mares in oestrus and dioestrus. In this study, uterine flush fluid samples were collected from eight reproductively healthy mares in either oestrus (n = 5) or dioestrus (n = 3). Proteomic analysis was performed using liquid chromatography‐tandem mass spectrometry. Of 172 proteins identified, six proteins (immunoglobulin lambda‐like polypeptide 1, haemoglobin subunit alpha, alpha‐1B‐glycoprotein, serotransferrin, apolipoprotein A‐1, and haemoglobin subunit beta) were significantly more abundant in oestrus. These proteins may contribute to the endometrial defence system through roles in inflammation, immunity or antimicrobial activity. In other species, some of these proteins have been described as immunoglobulins, negative acute phase proteins or defence agents against micro‐organisms. During dioestrus, immunoglobulin alpha‐1 chain C region‐related, complement factor I, CD 109 antigen and uterocalin, were significantly more abundant. Research in other species suggests that these four proteins contribute to the immune response through proposed immunoregulatory characteristics, complement system involvement or roles in B cell–T cell interactions. In conclusion, ten differentially abundant proteins were identified in the uterine flush fluid of mares in oestrus and dioestrus. Targeted studies on these proteins could elucidate their role in uterine defence mechanisms during the oestrous cycle in the mare.