Isolation and Differentiation of Mesenchymal Stem Cells From Bovine Umbilical Cord Blood

Currently, mesenchymal stem cells (MSCs) are used in veterinary clinical applications. Bone marrow and adipose tissue are the most common sources of stem cells derived from adult animals. However, cord blood which is collected non‐invasively is an alternative source of stem cells other than bone marrow and adipose tissue. Moreover, high availability and lower immunogenicity of umbilical cord blood (UCB) haematopoietic stem cells compared to other sources of stem cell therapy such as bone marrow have made them a considerable source for cell therapy, but MSCs is not highly available in cord blood and their immunogenicity is poorly understood. In this study, the cells with spindle morphology from 7 of 9 bovine UCB samples were isolated and cultured. These mesenchymal stromal cells were successfully differentiated to osteocytes, chondrocytes and adipocytes. In addition, Oct‐4 and SH3 were determined by RT‐PCR assay. It is the first report of isolation, culture, characterization and differentiation of bovine umbilical stem cells.     

Burst Conditions of Explosive Volcanic Eruptions Recorded on Microbarographs

Explosive volcanic eruptions generate pressure disturbances in the atmosphere that propagate away either as acoustic or as shock waves, depending on the explosivity of the eruption. Both types of waves are recorded on microbarographs as 1- to 0.1-hertz N-shaped signals followed by a longer period coda. These waveforms can be used to estimate burst pressures and gas concentrations in explosive volcanic eruptions and provide estimates of eruption magnitudes.     

Management of the exploitation of the lugworm Arenicola marina and the ragworm Nereis virens (Polychaeta) in conservation areas

• 1. In Europe, bait supply to anglers from the natural environment involves: (i) bait collection by anglers for own use, (ii) local semi-professional bait digging and (iii) professional bait digging contracted to distributors.
• 2. The management of bait digging activity is particularly acute where there is a conflict with conservation interests.
• 3. Two case studies are presented describing the population dynamics of: I Arenicola marina in the Lindisfarne nature reserve, and II Nereis virens in the Menai Strait; both in conservation areas.
• 4. Case I. Populations of A marina are subject to marked annual cycles in number and fresh weight. This case illustrates the response of the population to substantial overexploitation leading to a population crash. With the introduction of restrictive legislation the population was re-established by recruitment of adults from neighbouring unexploited populations.
• 5. Case II illustrates the dynamics of a N. virens population dominated by large old individuals; it does not represent a sustainable resource in contrast with populations of the same species in other locations.
• 6. The management of the exploitation of bait populations in conservation areas is discussed in relation to world patterns of utilization and supply.

     

Tumescent local anesthesia with ropivacaine in different concentrations in bitches undergoing mastectomy: plasma concentration and post‐operative analgesia

ObjectiveTo compare two concentrations of ropivacaine administered for tumescent local anesthesia (TLA) in dogs undergoing mastectomy.Study designProspective randomized clinical study.AnimalsSeventeen bitches of various breeds, aged 12 ± 2 years and weighing 10 ± 6.5 kg requiring total unilateral or bilateral mastectomy.MethodsDogs were premedicated with acepromazine (0.04 mg kg^?1) and morphine (0.4 mg kg^?1) intramuscularly. Anesthesia was induced with propofol (2.5 mg kg^?1) and midazolam (0.2 mg kg^?1) intravenously, followed by intubation and maintenance with isoflurane and TLA. Dogs were randomly allocated to receive TLA either with 0.1% ropivacaine (group G1) or with 0.05% ropivacaine (group G05). TLA was performed by insertion of a multihole needle under the skin and infusion of ropivacaine and lactated Ringer’s solution at a fixed volume of 15 mL kg^?1. Ropivacaine concentrations in arterial blood were measured by high-performance liquid chromatography. Post-operative pain was assessed using two scales (University of Melbourne pain scale and a modified composite measure pain scale) and von Frey filaments, 4 hours after TLA and at 1 hour intervals until sensitivity was regained. A score above 30% of the maximum possible score was considered a positive indicator of pain.ResultsPeak plasma concentrations of ropivacaine were measured 240 minutes after TLA in G1. Low concentrations were measured in G05 for 60 minutes, with subsequent increase. Analgesic rescue and return of sensitivity occurred at 7 ± 2.3 and 7 ± 1.9 hours (mean ± SD) after TLA for G1 and G05, respectively.Conclusions and clinical relevanceTumescent local anesthesia with ropivacaine provided satisfactory post-operative analgesia that lasted for several hours, with no difference in duration between the concentrations. No serious side effects were attributed to TLA. Results indicated that 0.05% ropivacaine provided adequate analgesia for mastectomy, however, more studies are required to support this conclusion.     

Experimental infection with BCG as a model of tuberculosis in the brushtail possum (Trichosurus vulpecula)

AIMS: To study the development and progression of lesions produced following experimental inoculations of possums with Bacille Calmette-Guérin (BCG) Pasteur Strain 1173P2 and to compare these with lesions that occurred following natural Mycobacterium bovis infection.

METHODS: Possums were inoculated with 5 x 10⁶ colony forming units (cfu) of BCG via the intra-dermal (I/D) route into the dorsum of the neck (n=38) or the left brachium (n=7), orally (n=10), via the endo-bronchial (E/B) route (n=12), or intravenously (I/V) (n=10, half of which received 5 x 10⁶ cfu and half of which received 5 x 10⁷ cfu of BCG). The possums were humanely killed between 1–4 weeks post inoculation (p.i.), and the nature and distribution of lesions examined grossly and histopathologically.

RESULTS: The distribution of lesions following I/D inoculation via either route was similar to that of the natural disease, but there were few lesions in the lung. Endo-bronchial inoculation resulted in pulmonary disease but produced few lesions outside the respiratory tract. Lesions produced by I/V inoculation were similar in distribution to those seen in terminally ill tuberculous possums. No lesions were produced following oral inoculation. Regression of lesions commenced after 3 weeks p.i.

CONCLUSIONS: Although the phenomenon of lesion resolution restricts the use of BCG to the study of early lesion development, it avoids the overwhelming disease induced using M. bovis and thus allows the early phases of the development and progression of tuberculosis in this species to be observed. Intra-dermal inoculation produced evidence that infection through the skin is associated with lesions in superficial lymph nodes, whereas pulmonary disease was associated with E/B inoculation. The results are consistent with the hypothesis that both percutaneous and respiratory routes are important in natural infection of possums with M. bovis.     

Accidental Mycobacterium bovis infection in a veterinarian

CASE: A veterinarian developed tenosynovitis and secondary carpal tunnel syndrome following accidental inoculation of Mycobacterium bovis during the necropsy of a tuberculous possum from Westland, New Zealand.

CLINICAL RELEVANCE: M. bovis infection is a zoonotic disease, and occupational exposure to tuberculous animals places people at risk of contracting the disease.

CONCLUSIONS: Adhering to safe work practices reviewed in this article is important to minimise the risk of infection to people handling tuberculous animals.     

Site of Intrauterine Artificial Insemination in the Bitch does not Affect Sperm Distribution within the Uterus

The aim of this study was to evaluate the distribution of frozen–thawed spermatozoa within the uterine lumen and oviducts following intrauterine laparoscopic deposition at two sites. Twelve bitches of unknown reproductive history were randomly distributed into two groups. Semen (3 ml containing 300 × 10⁶ frozen–thawed spermatozoa) was infused at the uterine body (UB group) or at the cranial tip of the left uterine horn. A 22‐G catheter was used to access the uterine lumen. Sperm cell distribution was evaluated after ovariohysterectomy performed 3 h after artificial insemination (AI). There was no difference between groups in mean time to perform AI. Spermatozoa were detected in all uterine segments, including the tip of both horns, but none was detected in the oviduct. The 22‐G catheter facilitated deposition of semen in the uterine lumen, particularly at the UB site. Sperm cell distribution occurred evenly along both horns, independent of the site of semen deposition.     

CLINICAL SIGNIFICANCE AND PROGNOSIS OF DEEP DIGITAL FLEXOR TENDINOPATHY ASSESSED OVER TIME USING MRI

Deep digital flexor (DDF) tendinopathy is one of the most frequent causes of foot lameness and the prognosis is guarded. The progress of lesion healing may be followed by magnetic resonance (MR) imaging to formulate a prognosis and to adapt the rehabilitation program. We assessed the correlation of outcome with total tendon damage and temporal resolution of MR abnormalities. Images from 34 horses with DDF tendinopathy that had undergone at least two low‐field standing MR examinations of the foot (mean 2.5 ± 1.3 times) were reviewed. No horse having a T1‐GRE hyperintense lesion over 30 mm in length or over 10% tendon cross‐sectional area returned to its previous activity level. Horses with concomitant lesions had worse outcome than horses with DDF tendinopathy only (P = 0.005). In all horses including those with excellent outcome, the lesion persisted, even mildly, in T1‐GRE and PD images. Horses with tendon lesion resolution on STIR‐FSE and T2‐FSE images on recheck examination had a better outcome (P = 0.0004 and P = 0.002, respectively), and all horses that returned to their previous level of performance had complete resolution of signal hyperintensity on the STIR‐FSE sequence. Although rehabilitation remains multifactorial, characteristics of DDF tendinopathy and concomitant lesions on first and recheck MR examinations allow refining the prognosis.