Feline corneal sequestrum: laboratory analysis of ocular samples from 12 cats

Feline corneal sequestrum is a common ocular condition typified by brown to black discoloration of the cornea. The nature of the discoloration has not been identified. The purpose of this study was to perform a laboratory investigation of ocular samples from 12 clinical cases of feline corneal sequestrum in an attempt to characterize the nature of the discoloration. The 12 cases were referred to the Ophthalmology Unit at the Animal Health Trust between April and September 2000, and were also part of a clinical review of 64 cases of feline corneal sequestrum described separately. Five laboratory techniques that are routinely performed at the Biomaterials Unit, Aston University were employed for analysis of the ocular samples. Ocular material included corneal sequestrum, tear samples, meibomian gland secretions, and bandage contact lenses from the 12 clinical cases. High-performance liquid chromatography data showed that total tear lipid in affected eyes was significantly lower than in control eyes (P = 0.016); total tear lipid in affected eyes was lower than in the unaffected, contralateral eyes of the same cat but the difference was not significant (P = 0.29). The presence of an unknown lipid class was observed in tears and meibomian secretions of affected, contralateral and control eyes. Scanning electron microscopy showed that the discoloration in affected corneas was not due to the presence of iron. Fluorescence spectroscopic analysis of sequestra, unaffected corneas and contact lenses (from affected and contralateral/unaffected eyes) showed that lipid and protein were present but did not play an important role in sequestra. Ultraviolet-visible light absorbance spectroscopy revealed a peak at 385 nm in unaffected corneas that was absent in sequestra and the difference was significant (P < 0.0001); this peak may be a characteristic feature of the normal feline cornea. The absorbance spectra displayed a peak at 280 nm in two sequestra suggesting that chromophore groups (e.g. melanin) were present. Optical microscopy performed on 10 sequestra revealed the presence of particles, which were consistent with the appearance of melanin particles, providing laboratory evidence that characterized the nature of the discoloration as melanin for the first time.     

Use of trichostatin A alters the expression of HDAC3 and KAT2 and improves in vitro development of bovine embryos cloned using less methylated mesenchymal stem cells

The aim of this work was to investigate the methylation and hydroxymethylation status of mesenchymal stem cells (MSC) from amniotic fluid (MSC‐AF), adipose tissue (MSC‐AT) and fibroblasts (FIB‐control) and to verify the effect of trichostatin A (TSA) on gene expression and development of cloned bovine embryos produced using these cells. Characterization of MSC from two animals (BOV1 and BOV2) was performed by flow cytometry, immunophenotyping and analysis of cellular differentiation genes expression. The cells were used in the nuclear transfer in the absence or presence of 50 nM TSA for 20 hr in embryo culture. Expression of HDAC1, HDAC3 and KAT2A genes was measured in embryos by qRT‐PCR. Methylation results showed difference between animals, with MSC from BOV2 demonstrating lower methylation rate than BOV1. Meanwhile, MSC‐AF were less hydroxymethylated for both animals. MSC‐AF from BOV2 produced 44.92 ± 8.88% of blastocysts when embryos were exposed to TSA and similar to embryo rate of MSC‐AT also treated with TSA (37.96 ± 15.80%). However, when methylation was lower in FIB compared to MSC, as found in BOV1, the use of TSA was not sufficient to increase embryo production. MSC‐AF embryos expressed less HDAC3 when treated with TSA, and expression of KAT2A was higher in embryos produced with all MSC and treated with TSA than embryos produced with FIB. The use of MSC less methylated and more hydroxymethylated in combination with embryo incubation with TSA can induce lower expression of HDAC3 and higher expression of KAT2A in the embryos and consequently improve bovine embryo production.     

An atypical Bacillus anthracis infection in a bull—A potential occupational health hazard

Bacillus anthracis infecting cattle is usually identified based on the typical symptom: sudden death. Bacillus anthracis causing atypical symptoms may remain undiagnosed and represent a potential occupational health hazard for, that is veterinarians and producers, butchers and tanners. In the year 2004, one case of sudden death in a dairy farm in southern Finland was diagnosed as bovine anthrax. Four years later 2008, an atypical case of anthrax was diagnosed in the same holding. The bull was taken to the Production Animal Hospital of the Faculty of Veterinary Medicine, University of Helsinki because of fever, loss of appetite and a symmetrically swollen scrotal sac. Penicillin treatment cured the fever but not the swollen scrotum. Before the intended therapeutic castration, a punctuate consisting of 10 ml fluid collected into a syringe from the scrotal sac was cultivated on blood agar at 37°C. After 24 hr, an almost pure culture of a completely non‐hemolytic Bacillus cereus‐like bacteria was obtained. The strain was identified as B. anthracis using Ba‐specific primers by the Finnish Food Safety Authority (RUOKAVIRASTO). After the diagnosis, the bull was euthanized and destroyed, the personnel were treated with prophylactic antibiotics and the clinic was disinfected. In this particular case, treatment with water, Virkon S and lime seemed to be effective to eliminate endospores and vegetative cells since no relapses of anthrax have occurred in 10 years. This case is the last reported anthrax case in Finland.     

The effectiveness of conservation measures to enhance nest survival in a meadow bird suffering from anthropogenic nest loss

The main reason for meadow bird declines is supposed to be the intensification of grassland management with earlier first harvest dates and more frequent harvests, resulting in high nest destruction rates. To increase productivity of meadow bird populations in intensified grassland areas a delay of mowing date and individual nest protection measures have been proposed. However, for ground-nesting songbirds such as the whinchat (Saxicola rubetra) the effectiveness of such measures remains widely unknown. In particular, if nest predation rate is high, measures to protect nests from agricultural destruction alone might be questionable. Here, we quantify whinchat nest survival of (1) unprotected nests situated in early mown meadows, (2) protected nests situated in early mown meadows, and (3) nests situated in late mown meadows. Analyses considered the fact that successful and unsuccessful nests are not found with equal probabilities. Periods of reduced nest survival were associated with mowing periods in the different types of meadows. In early mown meadows nest survival rates were low (S < 0.1), and both conservation measures, individual nest protection and delayed mowing, resulted in significantly increased nest survival rates (S > 0.7). Individual nest protection cannot avoid changes in habitat quality of intensively managed meadows, and therefore is only suitable as small-scale and short-term measure to increase nest success until a high portion of late mown meadows is established. Thus, we suggest that a combination of the two measures applied to intensified grassland fields should be provided to maintain viable sizes of endangered meadow bird populations.     

Drug Resistance in Equine Parasites: An Emerging Global Problem

Effective parasite control is essential for the maintenance of optimal health and performance in the horse. The worldwide escalation of parasite resistance is a major cause of concern for the horse industry. Parasite resistance to every main class of equine anthelmintic has been documented. Furthermore, dual and cross resistance also have been widely reported, despite different climatic and management practices seen throughout the world. Studies documenting parasite resistance to major classes of equine anthelmintics are discussed. Disagreement among researchers exists regarding how to effectively control equine internal parasites. Current theories of factors leading to resistance and control programs are discussed. It is clear that parasite resistance and control in the horse is an area requiring continued intensive study.     

Characterization of experimental Mycoplasma gallisepticum infection in captive house finch flocks

The use of controlled, horizontal-transmission experiments provides detailed information on the spread of disease within fixed social groups, which informs our understanding of disease dynamics both in an empirical and theoretical context. For that reason, we characterized in 2002, horizontal transmission of Mycoplasma gallisepticum (MG) in two flocks of 11 wild-caught house finches housed in outdoor aviaries over a 6-mo period. All birds were initially free of MG by a polymerase chain reaction (PCR)-based test, rapid plate agglutination (RPA), and the scoring of physical signs. We inoculated one flock member bilaterally in the palpebral conjunctiva and reintroduced it into its cage. Index birds developed conjunctivitis within 3 to 5 days but died 13 and 20 days postinfection (PI) possibly because of very severe weather. The proportion of birds with physical signs increased gradually, reached 40% at 6 wk PI, and fluctuated around 40% until 21 wk PI. By the time our experiment ended at 24.5 wk PI, 28% of the birds still exhibited physical signs. Across both flocks, 80% of the birds developed unilateral or bilateral conjunctivitis, and several birds relapsed. The appearance of physical signs in new individuals occurred between 10 and 144 days PI (median 41 days PI). Physical signs lasted 1-172 days (median 42 days). Birds that became infected earlier during the experiment developed more severe conjunctivitis, and there was a tendency for birds that developed bilateral conjunctivitis to develop physical signs earlier. Most birds that developed physical signs of MG were also PCR- and RPA-positive, although we detected a single asymptomatic carrier and a single symptomatic false negative. No birds died as a result of secondary MG infection.     

Biomechanical Analysis of Unilateral External Skeletal Fixators Combined with IM-Pin and Without IM-Pin Using Finite-Element Method

OBJECTIVES: To determine if a unilateral external skeletal fixator (ESF) with a carbon fiber connecting rod (IMEX SK) without an intramedullary (IM)-pin is mechanically comparable with a unilateral ESF with a stainless-steel connecting rod (IMEX KE) with an IM-pin. STUDY DESIGN: Finite-element method (FEM)-computer simulation. METHODS: FEM models were validated by comparison against data from mechanical testing. Three-dimensional FEM models of a femur with a mid-diaphyseal fracture with a 20 mm gap were developed with 4 unilateral external skeletal fixator devices (6-pin KE, 6-pin KE IM-pin, 6-pin SK, and 6-pin SK IM-pin). A 300 N load was applied to the femur at the proximal end in a direction of theta = 10 degrees distally and phi = 10 degrees laterally cranially. Relative displacements in x-, y- and z-directions at the gap were obtained and the overall stiffness was calculated as 300 N/total displacement. Load transfer at the pin-bone interface (PBI) was assessed by determining the von Mises stress maxima at the PBI-related nodes. RESULTS: The 6-pin SK had superior mechanical performance compared with the 6-pin KE by exhibiting smaller displacements in all directions and higher stiffness. Compared with the 6-pin KE IM-pin, the 6-pin SK (without IM-pin) was superior in craniocaudal and lateromedial displacements, but inferior in axial displacements, overall stiffness and von Mises stress maxima. The 6-pin SK IM-pin was superior to the 6-pin KE IM-pin based on smaller displacements and higher stiffness. CONCLUSIONS: Although the SK device had superior mechanical performance compared with a KE device in a unilateral configuration, the addition of an IM-pin continues to be a powerful method of enhancing mechanical performance of either IMEX SK or IMEX KE unilateral constructs in clinical cases. CLINICAL RELEVANCE: Based on the results of this FEM study we recommend the use of the "tied-in" IM-pin with the ESF clinically when striving for high rigidity. In less challenging situations, a unilateral SK ESF without IM-pin might provide sufficient rigidity for a successful fracture repair.