Vascular ultrasonographic findings in canine patients with clinically diagnosed phlebitis

Peripheral vein phlebitis (inflammation) is a relatively frequent complication in dogs, however, published information on the ultrasonographic characteristics is currently lacking. This prospective, observational study describes the ultrasound (US) characteristics of normal canine cephalic veins, and veins with clinical phlebitis. Correlations among US findings and between US findings versus time that the intravenous catheter was in place were investigated. Safety of the US procedure was evaluated. Fifty patients were prospectively recruited for the study and 18 met the final inclusion criteria. Each patient underwent daily US examinations and was assessed for multiple criteria (vascular wall appearance, compressibility, spontaneity of flow, color fill, and presence/absence of filling defects, flow contour, direction, non‐pulsatility). Characteristics of normal canine cephalic veins were as follows: smooth and thin wall, complete compressibility, no flow disturbances, no filling defects, smooth flow contours, and unidirectional, non‐pulsatile flow with no turbulence. Characteristics of cephalic veins with clinical phlebitis were as follows: wall thickening (83%), decreased compressibility (55%), filling defects consistent with intraluminal thrombus (55%), vessel wall hyperechogenicity (44%), and abnormal color Doppler flow (39%). Significant correlations were found between Doppler filling defects and compressibility, Doppler filling defects and presumed thrombosis, and compressibility and presumed thrombosis (P = .001, P = .001, P = .000, respectively). No correlation was found between the US findings and time the intravenous catheter was in place. Findings indicated that duplex and compressibility US are feasible and safe methods for characterizing and monitoring cephalic veins in dogs with clinical phlebitis.     

Dexmedetomidine constant rate infusion for 24 hours during and after propofol or isoflurane anaesthesia in dogs

OBJECTIVE: To evaluate cardiovascular and respiratory effects and pharmacokinetics of a 24-hour intravenous constant rate infusion (CRI) of dexmedetomidine (DMED) during and after propofol (PRO) or isoflurane (ISO) anaesthesia in dogs. STUDY DESIGN: Prospective, randomized, cross-over study. ANIMALS: Ten healthy adult Beagles. METHODS: Instrumented dogs received a DMED-loading bolus (25 microg m(-2)) at time 0 followed by a 24-hour CRI (25 microg m(-2) hour(-1)), with PRO or ISO induction/maintenance of anaesthesia during the first 2 hours (PRO and ISO treatment groups, respectively). Cardiovascular, respiratory, blood gas, airway gas, serum chemistry variables and DMED plasma concentration data were collected at -15, 5, 15, 30, 45, 60, 90 and 120 minutes. A number of cardiorespiratory and tissue oxygenation variables were calculated from the above data. After the 2-hours of anaesthesia, heart and respiratory rates and electrocardiograms were recorded and DMED plasma concentrations were determined for up to 26 hours. RESULTS: Vasopressor effects and the decrease in heart rate (HR) and cardiac index induced by DMED were greater for PRO than ISO, but were within clinically acceptable ranges. Adequate oxygenation was maintained above the critical O(2) delivery level. The overall incidence of unfavourable arrhythmias was low and tended to vary inversely with HR. Mean DMED plasma concentration ranged from 0.23 to 0.47 ng mL(-1) for both groups during the 24-hour CRI with a mean elimination half-life of approximately 0.46 hour. CONCLUSION AND/CLINICAL RELEVANCE: DMED CRI resulted in typical alpha(2)-agonist induced haemodynamic changes with minimal respiratory effects, and appeared to be an efficacious adjunct during and after PRO or ISO anaesthesia in healthy dogs.     

Agreement between ELISA and complement fixation test used for diagnosing of paratuberculosis in goats

An ELISA with a lipoarabinomannan as an antigen, developed for diagnosis of bovine paratuberculosis, has been adapted for use in goats, and compared with complement fixation test. Kappa value of 0.62 indicated good agreement between CFT and the adapted ELISA and proved that the investigated ELISA may be helpful in diagnosis of Mycobacterium avium subsp. paratuberculosis infection in goats. The ELISA has been used to screen a randomly selected representative sample of Polish breeding goat population (21.78% of herds, 21.33% of goats). It has been demonstrated that only 2.42% of animals coming from 15.79% of herds were seropositive. Within-herd seroprevalence varied from 1.69% to 38.10%. Most of the infected animals (67.07%) were 3- 4-years-old. No seropositive cases were found in group up to 1-year-old animals.     

Feline hip joint anatomy in magnetic resonance images

The aim of this study was to develop an anatomical model of the feline hip joint for low‐field magnetic resonance imaging (LF‐MRI) based on high‐field magnetic resonance imaging (HF‐MRI). The study was performed on six adult clinically healthy European shorthair cats, aged 1–3 years, with body weight of 2.8–4.4 kg. The animals were examined with the use of the Vet‐MRI Grande Esaote LF (0.25 T) scanner and high‐field Siemens Magnetom TRIO (3 T) MRI scanner. In the LF‐MRI, most satisfactory results in T1‐weighted images were obtained when TE was 26 ms in all three planes and when TR was 350–950 ms in the transverse plane, 950–1150 ms in the sagittal plane and 520–750 ms in the dorsal plane. In T2‐weighted images, TE was 90 ms in the transverse and dorsal plane and 120 ms in the sagittal plane. The results were presented as images acquired with LF‐MRI scanners in three planes. The slice thickness was 3 mm for each plane. In LF‐MRI, muscles in the hip joint region and round ligament were well visualized. Unlike in LF‐MRI, the cross section of the femoral nerve was identified in HF‐MRI scans. In examinations of the feline hip joint, the main limitations of LF‐MRI were a lack of reliable contrast between articular cartilage and synovial fluid as well as longer scan time. Despite the above, LF‐MRI images were characterized by good contrast between bones and the surrounding soft tissues.     

Comparison of the intravenous and intravaginal route of oxytocin administration for cervical dilation protocol and non‐surgical embryo recovery in oestrous‐induced Santa Inês ewes

This study compared the effects of intravaginal and intravenous routes of oxytocin (OT) administration in 46 oestrous‐induced Santa Inês ewes (6‐day treatment with progestin‐releasing intravaginal sponges and a single injection of 200 IU of eCG at the time of sponge removal) that underwent transcervical embryo recovery 6–7 days after oestrous onset and mating. All ewes received 37.5 μg of d‐cloprostenol via latero‐vulvar route, and 1 mg of oestradiol benzoate i.m. 16 hr before and 50 IU of OT 20 min before non‐surgical embryo recovery (NSER), with OT being administered intravenously (n = 21) or intravaginally (n = 21). An overall oestrous response was 95.6% (44/46), and adequate cervical retraction could be accomplished in 78.6% (33/42) of ewes. The percentage of successful NSER procedures was 57% (24/42) or 72.7% (24/33) of animals with sufficient cervical retraction. The duration of NSER procedure averaged 28 min (range: 17–40 min) and ~96% of flushing fluid could be recovered (range: 85%–100%). Out of 18 ewes that could not undergo NSER, 12 (66.6%) presented various anatomical barriers, whilst the other 33.4% did not present these barriers and still could not be traversed. Excluding the ewes with those anatomical features, the overall success rate of NSER was 80% (24/30). The route of OT administration had no effect on NSER efficiency or the ease with which transcervical embryo flushing was performed. Both routes of OT administration can be used for cervical dilation protocol. Discarding ewes with anatomical features precluding cervical penetration is highly recommended to increase the efficacy of NSER in sheep.     

Decreases in urine specific gravity and urinary creatinine in dogs with different diseases but with normal renal function

The subject of this study was to determine urine specific gravity (USg) and urinary creatinine (UCrn) in dogs with different diseases but with normal renal function. Sick dogs with different diseases were divided into nine groups. Dogs suffering from polyuria/polydipsia, vomits, diarrhoea and females in oestrus or pregnant were excluded from the studies. The healthy dogs served as a control group. Over a three-year period, a total of 267 dogs were examined clinically as well as using imaging and laboratory diagnostics methods. In sick dogs, USg and UCrn were found to be essentially decreased (except animals with neurological and uterine diseases, and neurological diseases, respectively), as compared with normal dogs. In clinically healthy animals of the control group, UCrn and USg did not significantly differ between the females and males. As for the control group, no correlation between UCrn/USg and the body weight/age was found, either.     

A simple and efficient CRISPR/Cas9 platform for induction of single and multiple,heritable mutations in barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L.)

Background

Genome editing of monocot plants can be accomplished by using the components of the CRISPR/Cas9 (clustered regularly interspaced short palindromic repeat/CRISPR associated Cas9) technology specifically optimized for these types of plants. Here, we present the development of RNA-guided Cas9 system for simplex and multiplex genome editing in barley.

Results

We developed a set of customizable RNA-guided Cas9 binary vectors and sgRNA modules for simplex and multiplex editing in barley. To facilitate the design of RNA-guided Cas9 constructs, the pBract derived binary vectors were adapted to Gateway cloning and only one restriction enzyme was required for construction of the sgRNA. We designed a synthetic, codon optimized Cas9 gene containing the N terminal SV40 nuclear localization signal and the UBQ10 Arabidopsis 1st intron. Two different sgRNAs were constructed for simplex editing and one polycistronic tRNA-gRNA construct (PTG) for multiplex editing using an endogenous tRNA processing system. The RNA-guided Cas9 constructs were validated in transgenic barley plants produced by Agrobacterium-mediated transformation. The highest mutation rate was observed in simplex editing of the cytokinin oxidase/dehydrogenase HvCKX1 gene, where mutations at the hvckx1 locus were detected in 88% of the screened T₀ plants. We also proved the efficacy of the PTG construct in the multiplex editing of two CKX genes by obtaining 9 plants (21% of all edited plants) with mutations induced in both HvCKX1 and HvCKX3. Analysis of the T₁ lines revealed that mutations in the HvCKX1 gene were transmitted to the next generation of plants. Among 220 screened T₁ plants we identified 85 heterozygous and 28 homozygous mutants, most of them bearing frameshift mutations in the HvCKX1 gene. We also observed independent segregation of mutations and the Cas9-sgRNA T-DNA insert in several T₁ plants. Moreover, the knockout mutations of the Nud gene generated phenotype mutants with naked grains, and the phenotypic changes were identifiable in T₀ plants.

Conclusions

We demonstrated the effectiveness of an optimized RNA-guided Cas9 system that can be used for generating homozygous knockout mutants in the progeny of transgenic barely plants. This is also the first report of successful multiplex editing in barley using a tRNA processing system.

     

Occurrence of reproductive disorders in pig herds with and without Chlamydia suis infection – statistical analysis of breeding parameters

Chlamydiae are frequently encountered Gram‐negative intracellular eubacteria that can cause clear manifestations or clinically asymptomatic disorders. C. suis and other chlamydia are primarily isolated in cases of reproductive disorders. This study was performed to estimate the impact of Chlamydia suis infection on reproduction in sows by analyzing reproduction rates and breeding parameters. The test was conducted on first generation (F1) pigs from Polish Landrace (PL) × Polish Large White (PLW). Sixty‐four herds were investigated and 500 vaginal swabs were collected. Isolation of DNA was carried out directly from the swabs. All samples were analyzed for Chlamydia suis by real‐time PCR with a locked nucleic acid (LNA)‐containing probe. To analyze the impact of chlamydia infection on reproductive parameters, evaluation questionnaires were used. Reproductive problems were found in 77.3% of the farms tested. The receiver operating characteristic curve indicated that in the farms with 10 up to 120 sows, there were higher reproductive problems with chlamydia infection than in smaller and bigger pig farms. The most common problems were estrus repetition, which was reported by 57.81% of the surveyed farms, and the birth of dead piglets, which was reported by 31.25% of the investigated pig farms. Abortions, which were reported by 28.12% of the surveyed farms, were the least common reproductive disorders.     

Identification of genome‐wide selection signatures in the Limousin beef cattle breed

The study is aimed at identifying selection footprints within the genome of Limousin cattle. With the use of Extended Haplotype Homozygosity test, supplemented with correction for variation in recombination rates across the genome, we created map of selection footprints and detected 173 significant (p < 0.01) core haplotypes being potentially under positive selection. Within these regions, a number of candidate genes associated inter alia with skeletal muscle growth (GDF15, BMP7, BMP4 and TGFB3) or postmortem proteolysis and meat maturation (CAPN1 and CAPN5) were annotated. Noticeable clusters of selection footprints were detected on chromosomes 1, 4, 8 and 14, which are known to carry several quantitative trait loci for growth traits and meat quality. The study provides information about the genes and metabolic pathways potentially modified under the influence of directional selection, aimed at improving beef production characteristics in Limousin cattle.