Expression of equine morbillivirus (EMV) matrix and fusion proteins and their evaluation as diagnostic reagents

Full-length cDNA clones coding for the matrix (M) and fusion (F) proteins of equine morbillivirus (EMV) were isolated by RT-PCR, and expressed in Escherichia coli using two different expression systems. Western blot analysis indicated that the M and F proteins, expressed either by itself or as fusion proteins with glutathione S-transferase (GST), were insoluble and degraded after expression. Analysis of the degradation pattern of recombinant M protein suggested that the N-terminus of the matrix protein might be more stable and antigenic than the C-terminal region. Therefore a third system was used to express a truncated M protein, composed of the N-terminal amino acid residues 1-197, with a (His)6-tag attached at the N-terminus. This recombinant protein [(His)6-Mtr], was stable but was also insoluble. After one-step affinity purification under denaturing conditions, (His)6-Mtr was used to monitor the antibody response to EMV infection by Western blot and ELISA. We obtained a 100% correlation between Western blot and virus neutralisation testing although the number of positive sera available for testing was very limited, which included seven horse, two rabbit and one human sera.     

Demonstration of intraocular leptospira in 4 horses suffering from equine recurrent uveitis (ERU)

Vitreous samples from 43 horses which underwent vitrectomy because of equine recurrent uveitis (ERU) were cultured for leptospires. Out of 4 vitreous samples (4/43 = 9%), leptospires could be isolated. In 3 cases, serovar grippotyphosa, and in one case, a serovar out of the serogroup Australis were identified. So for the first time, in several horses with ERU in vivo cultures of vitreous material were positive for leptospires. A strong evidence of association between leptospiral infection and uveitis is discussed for many years. In this investigation the leptospiral etiology is confirmed. Vitreous material from 42 and serum samples from 40 horses were tested for specific antibodies to leptospira by microagglutination test (MAT). In 34 vitreous samples (34/42 = 81%), leptospiral antibody titers of 1:50 or higher were detected. In 33 horses (33/40 = 83%) leptospiral antibody titers of 1:50 or higher could also be detected in the serum. Altogether, leptospiral antibodies were detected by the MAT in the serum and in the vitreous material of 39 of 43 horses (= 91%) subjected to vitrectomy. These results indicate, that ERU is probably often a sequel to systemic Leptospira interrogans infection. The presence of intact leptospires and specific antibodies in eyes affected with ERU indicates a local antibody production to leptospira organisms and/or their antigens.     

Attack of black flies (Diptera, Simuliidae) on horses in Basel (Switzerland)

In April 1996 an attack of black flies (Diptera, Simuliidae) to horses was observed in the region of Basle, Switzerland. Females of Simulium erythrocephalum and Simulium ornatum were involved in this attack, part of them had taken a blood meal. For S. ornatum this is the first record in Switzerland that this species takes blood meals on horses.     

Babesia sp. in Colombian bats (Microchiroptera)

Two leaf-chinned bats (Mormoops megalophylla) collected in 1963 in central Colombia were heavily infected with Babesia sp., probably Babesia vesperuginis. Both bats had pronounced splenomegaly. This is the first report of a Babesia sp. infection of a bat in the Americas.     

Transmission electron microscopy studies of (111) twinned silver halide microcrystals

Minimal information is available on the autonomic response to exercise under adverse environmental conditions. Traditionally, pharmacological blockade has been used to study autonomic responsiveness but, owing to its invasive nature, such studies have been limited in their scope. Recent advances in electrocardiographic tape recording, telemetry and associated computing systems have provided investigators with noninvasive methods for assessing the autonomic response to various physiological stressors. This article describes methods for the analysis of heart rate variability (HRV) and discusses the reports of those who have used HRV analysis to evaluate autonomic regulation during exercise, heat exposure and the combination of these 2 stressors. Spectral analysis of HRV reduces variations in the R-R interval into component sine waves of differing amplitude and frequency. Amplitude (variance) is displayed as a function of frequency, and power (cumulative variance) is calculated for specified frequency ranges (< 0.03 Hz, 0.03 to 0.15 Hz and 0.15 to 0.5 Hz). Parasympathetic nervous system activity can be inferred from the several indices of high frequency power; however, the estimation of sympathetic nervous system activity from low frequency power is more problematic. Data on HRV have shown that sympathovagal regulation during exercise is dependent on the intensity of the activity and the environmental conditions. At the onset of exercise, heart rate is increased by a reduction in vagal tone and a temporary increase in sympathetic tone. A continuation of physical activity is associated with a continued withdrawal of vagal activity and an attenuation of sympathetic nervous system tone. However, with the additional stimulus of a heated environment, sympathetic activity remains increased throughout exercise.     

Plasma alpha-tocopherol, beta-carotene, and lipid levels in semi-free-ranging Przewalski horses (Equus przewalskii)

To describe the features of railway-related deaths in Cape Town, South Africa, we reviewed demographic, autopsy, and accident report data on all such deaths between 1 April 1992 and 30 September 1994. Of the 379 railway-related deaths, 27 were among pedestrians or commuters who were hit by a train while crossing the track, 38 were among commuters who fell from moving trains, 32 were suicides, 43 were the result of criminal violence on trains or at railway stations, and 38 were due to other causes. Most railway fatalities were among men between the ages of 25 and 44 years. About half of all railway fatalities occurred at peak commuting times, with high levels of violence (often robbery related) recorded during the evening peak. A blood alcohol concentration > 0.1 g/100 ml was found in 35% of the people who died from crossing the track or falling from moving trains. Fatal railway injury is characterized by extensive disruption of more than one body region. The high levels of fatal railway injury make a strong case for a range of injury control interventions, including ticket control, surveillance, law enforcement, and safety engineering.     

Equine monocyte-derived macrophage cultures and their applications for infectivity and neutralization studies of equine infectious anemia virus

Equine infectious anemia virus (EIAV) has been shown to infect cells of monocyte/macrophage lineage. These primary cells are intrinsically difficult to obtain, to purify and to culture in vitro for extended periods of time. As a result, most in vitro studies concerning this lentivirus make use of primary equine fibroblasts or transformed canine or feline cell lines. We describe methods that yield reproducibly pure cultures of equine blood monocytes from peripheral blood mononuclear cells. The in vitro differentiation of these cells into mature equine macrophage was verified using various cytochemical staining methods. The equine monocyte-derived macrophage (MDM) cultures were found to replicate cell-adapted and field strains of EIAV more efficiently than cultures of fully differentiated equine splenic macrophage. Having established reproducible and fully differentiated cultures of equine macrophage, in vitro assays of virus infectivity and serum neutralization were developed using the in vivo target cell of EIAV. These procedures, while developed for the EIAV system, should be equally useful for in vitro cultures of other macrophage-tropic pathogens of horses.     

Serologic studies of domestic cats for potential human pathogenic virus infections from wild rodents]

For several viral infections a reservoir in wild rodents has been demonstrated. Some of the agents are known or suspected to be pathogenic for humans. Because improvements in hygiene have reduced direct human contact with rodents, domestic cats could be acting as active transmitters of these viruses from rodents to man. We selected 4 such pathogens--ortho- and parapox-, hanta- and encephalomyocarditis viruses--which, in different ways, may lead to serious human illness: Ortho- and parapoxvirus infections may cause localized pox lesions following direct skin contact. In general, the lesions heal without complications; in immunosuppressed or -deficient individuals, however, infection may generalize and take a dramatic course. Hantaviruses exist in various serotypes with different pathogenicity for human beings, varying from asymptomatic infection to highly fatal disease. In central and northern Europe the Puumala serotype is predominant causing influenza-like symptoms and renal dysfunction. Human infections arise from inhalation of aerosolized excreta of persistently infected rodents. Infections of man associated with encephalomyocarditis virus were demonstrated sporadically in cases of encephalitis and meningitis. In the present study, we investigated in 200 feline serum samples the prevalence of antibodies to ortho- and parapox-, hanta- and encephalomyocarditis virus. All serum samples were from cats that had been allowed to roam outside and to hunt. They were submitted from all parts of Austria for routine diagnosis in 1993. Four per cent of cats showed antibodies to orthopoxviruses with haemagglutination inhibition (HI) titres of 16-512; because of extensive cross-reactivity, positive samples reacted with all investigated orthopoxviruses (a feline orthopoxvirus recently isolated in Vienna, the reference strain of cowpox virus, Brighton, and vaccinia virus, strain IHD), only varying in titre. The specificity of the results was confirmed by virus neutralisation (VN) test, in which the same sera showed titres of 4-32. These data imply that, at least in Austria, unrecognized or subclinical orthopoxvirus infection in cats is more common than previously thought. In contrast to orthopoxviruses, all serum samples proved negative to parapoxvirus (parapoxvirus bovis 1) in VN test. In the same 200 samples, a seroprevalence of 5% was found to hantavirus (immunofluorescence antibody assay), indicating that domestic cats are susceptible to this virus and that infection is not uncommon in cat populations. Because higher titres were obtained against the Puumala serotype compared to the more pathogenic serotype Hantaan, it is most likely that the cats had experienced Puumala infections. Using HI test, antibodies to encephalomyocarditis virus (EMCV) were demonstrated in only 1.5% of the feline serum samples; although the antibody titres were low (16 and 32, respectively) we consider them specific, because these sera proved positive in VN test as well. Nevertheless, EMCV infection in domestic cats seems to be of low importance. The serological results presented in this paper, together with virological and epidemiological data, indicate that the domestic cat plays an important role only in the transmission of orthopoxviruses to human beings, but not in the case of parapox-, hanta-, and encephalomyocarditis virus.     

Exercise studies in horses: 2. The cardiac response to exercise in normal horses and in horses with chronic obstructive pulmonary disease

The relationship of velocity (v) to heart rate (HR)and of kinetic energy (KE) to heart rate, were investigated in 6 normal horses and in 6 horses with chronic obstructive pulmonary, disease (COPD). Radiotelemetry was used to determine the heart rate while subjects were ridden on a 400m track. Velocity was measured by stop-watch between 2 markers 50 m apart at the end of the track. Kinetic energy was calculated from the formula KE =1/2 Mv2, where M = mass of horse, rider, saddle and bridle (KG) and V = velocity (metres per second). In all subjects, the relationship of velocity to heart rate was a linear one at each individual pace (i.e. walk, trot and gallop). There were however significant differences between the v/HR regression curves of each pace. These differences indicated that the increase in velocity per unit increase in heart rate was greater at the trot than at the walk, and greater at the gallop than at the trot. When data for all paces were combined, the relationship log v/log HR was a linear one. The above findings were also true of the relationship of kinetic energy to the heart rate. In horses, which reached maximal heart rate at a relatively low velocity, the v/HR curve became asymptotic. At comparable submaximal velocities, the heart rate of COPD subjects was approximately 20 beats per minute higher than that of normal subjects. This was also true when the term "kinetic energy per units mass" was substituted for velocity.     

Isolation of Listeria monocytogenes in slaughter animals and bats of Togo (West Africa)

Listeria monocytogenes was isolated from the intestinal lymph nodes of 8 out of 342 apparently healthy slaughter animals at the municipal abattoir in Lome (Togo) and from the faeces of 3 out of 236 bats. Except two strains of serovar 4b isolated from bats, all the nine others belonged to serovar 1/2a. The positive findings of slaughter animals were made from the carcasses of 2 pigs, 5 little ruminants and 1 cattle. The findings are discussed in the light of present knowledge on the occurrence of L. monocytogenes in some species of animals and its implications for the human being.     

Biological characterization of Rev variation in equine infectious anemia virus

Sequence analysis identified significant variation in the second exon of equine infectious anemia virus (EIAV) rev. Functional analysis indicated that limited amino acid variation in Rev significantly altered the export activity of the protein but did not affect Rev-dependent alternative splicing. EIAV Rev can mediate export through two independent cis-acting Rev-responsive elements (RREs), and differences among Rev variants were more pronounced when both RREs were present. Variation in Rev may be an important mechanism for regulation of virus replication in vivo and may contribute to changes in clinical disease.     

Vector competence of mosquitoes (Diptera:Culicidae) from Massachusetts for a sympatric isolate of eastern equine encephalomyelitis virus

This study examined the characteristics of students with specific learning disabilities in either reading and spelling or arithmetic. Based on scores obtained using the revised Woodcock-Johnson Psycho-Educational Battery, students with a marked weakness in arithmetic relative to reading and spelling were designated as Group A. Group R-S showed the opposite pattern. Each group included 30 participants ranging in age from 7 to 16 years, with a mean age of 10 years. The boy-to-girl ratios were 16:14 and 19:11 in Group A and Group R-S, respectively. Comparisons using measures from the Wechsler Intelligence Scale for Children-Third Edition (WISC-III) indicated that Group A was weaker in nonverbal skills than Group R-S, despite equivalent overall IQ scores between the two groups. Group R-S showed a within-group strength in nonverbal versus verbal skills. Group A students were more likely than Group R-S students to have counseling provided as part of their Individualized Education Program, suggesting greater socioemotional difficulty among Group A students. The present study supports the connection between nonverbal skills and socioemotional functioning noted by previous researchers, and generalizes findings from earlier studies to more current test editions.     

Skin potential activity in rats, cats, and primates (including man): A phylogenetic point of view.

Measured skin potential activity under waking conditions in 2 experiments with a total of 60 rats, 21 cats, 40 nonhuman primates, and 87 normal adult humans. In agreement with findings of previous researchers, skin potential response waveform was always monophasic negative in rats and cats, but in humans it took 3 forms. By contrast, it was always monophasic positive in simian nonhuman primates, although prosimiae gave monophasic negative waves. A relationship between skin potential level and skin potential response could not be observed in any S except humans. From these results, an attempt was made to relate skin potential activity to the peripheral mechanism involved in these species based on a phylogenetic viewpoint. (20 ref) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Hoary alyssum (Berteroa incana) toxicity in a herd of broodmare horses

We report the case of a 71-year old male patient with a chronic lymphoid leukaemia of 10 years' duration who abruptly suffered deterioration of this general condition and haematological disturbances suggesting worsening of this blood disease (Richter's syndrome). Blood cultures demonstrated a Campylobacter coli septicaemia. Treatment with antibiotics resulted in a return to the previous clinical and biological situation. The various visceral manifestation of the infection are discussed.