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Evolutionary history of MHC class I genes in the mammalian order Perissodactyla.

We carried out an analysis of partial sequences from expressed major histocompatibility complex (MHC) class I genes isolated from a range of equid species and more distantly related members of the mammalian order Perissodactyla. Phylogenetic analysis revealed a minimum of six groups, five of which contained genes and alleles that are found in equid species and one group specific to the rhinoceros. Four of the groups contained only one, or very few sequences, indicating the presence of relatively nonpolymorphic loci, while another group contained the majority of the equid sequences identified. These data suggest that a diversification of MHC genes took place after the split between the Equidae and the Rhinocerotidae yet before the speciation events within the genus Equus.

Animals↗

Phylogenetics of Perissodactyla and tests of the molecular clock.

Two mitochondrial genes, the protein-coding cytochrome c oxidase subunit II (COII) gene and a portion of the 12S rRNA gene, were used for phylogenetic investigation of the mammalian order Perissodactyla. The primary objective of the study was to utilize the extensive fossil record of perissodactyls for calibrating molecular clocks and comparing estimates of divergence times using both genes and two fossil calibration points. Secondary objectives included clarification of previously unresolved relationships within Tapiridae and comparison of the results of separate and combined analyses of two genes. Analyses included several perissodactyl lineages representing all three families (Tapiridae, Equidae, and Rhinocerotidae), most extant genera, all four species of tapirs, two to four species of rhinoceros, and two species of Equus. The application of a relatively recent fossil calibration point and a relatively ancient calibration point produced greatly different estimates of evolutionary rates and divergence times for both genes, even though a relative rates test did not find significant rate differences among taxa. A likelihood-ratio test, however, rejected a molecular clock for both genes. Neither calibration point produced estimates of divergence times consistent with paleontological evidence over a range of perissodactyl radiations. The combined analysis of both genes produces a well-resolved phylogeny with Perissodactyla that conforms to traditional views of interfamilial relationships and supports monophyly of neotropical tapirs. Combining the data sets increases support for most nodes but decreases the support for a neotropical tapir clade because the COII and 12S rRNA data sets are in conflict for tapir relationships.

Animals↗

Plasma disposition and faecal excretion of oxfendazole, fenbendazole and albendazole following oral administration to donkeys.

Fenbendazole (FBZ), oxfendazole (fenbendazole sulphoxide, FBZSO), and albendazole (ABZ) were administered orally to donkeys at 10mg/kg bodyweight. Blood and faecal samples were collected from 1 to 120 h post-treatment. The plasma and faecal samples were analysed by high performance liquid chromatography (HPLC). The parent molecule and its sulphoxide and sulphone (FBZSO(2)) metabolites did not reach detectable concentrations in any plasma samples following FBZ administration. ABZ was also not detected in any plasma samples, but its sulphoxide and sulphone metabolites were detected, demonstrating that ABZ was completely metabolised by first-pass mechanisms in donkeys. Maximum plasma concentrations (C(max)) of FBZSO (0.49microg/mL) and FBZSO(2) (0.60microg/mL) were detected at (t(max)) 5.67 and 8.00h, respectively, following administration of FBZSO. The area under the curve (AUC) of the sulphone metabolite (10.33microg h/mL) was significantly higher than that of the parent drug FBZSO (5.17microg h/mL). C(max) of albendazole sulphoxide (ABZSO) (0.08g/mL) and albendazole sulphone (ABZSO(2)) (0.04microg/mL) were obtained at 5.71 and 8.00h, respectively, following ABZ administration. The AUC of the sulphoxide metabolite (0.84microg h/mL) of ABZ was significantly higher than that of the sulphone metabolite (0.50microg h/mL). The highest dry-faecal concentrations of parent molecules were detected at 32, 34 and 30h for FBZSO, FBZ and ABZ, respectively. The sulphide metabolite was significantly higher than the parent molecule after FBZSO administration. The parent molecule was predominant in the faecal samples following FBZ administration. After ABZ administration, the parent molecule was significantly metabolised, probably by gastrointestinal microflora, to its sulphoxide metabolite (ABZSO) that showed a similar excretion profile to the parent molecule in the faecal samples. The AUC of the parent FBZ was significantly higher than that of FBZSO and ABZ in faeces. It is concluded that the plasma concentration of FBZSO was significantly higher than that of FBZ and ABZ. Although ABZ is not licensed for use in Equidae, its metabolites presented a greater plasma kinetic profile than FBZ which is licensed for use in horses. A higher metabolic capacity, first-pass effects and lower absorption of benzimidazoles in donkeys decrease bioavailability and efficacy compared to ruminants.

Administration, Oral↗

African horse sickness in Portugal: a successful eradication programme.

African horse sickness (AHS) was diagnosed for the first time in southern Portugal in autumn 1989, following outbreaks in Spain. AHS virus presence was confirmed by virus isolation and serotyping. An eradication campaign with four sanitary zones was set up by Central Veterinary Services in close collaboration with private organizations. Vaccination began on 6 October. In February 1990, vaccination was extended to all Portuguese equines (170000 animals). There were 137 outbreaks on 104 farms: 206 of the equidae present died (16%) or were slaughtered (14%); 81.5% were horses, 10.7% were donkeys and 7.8% were mules. Clinical AHS occurred more frequently in horses than donkeys and mules. In the vaccinated population, 82 animals (62.2% horses and 37.8% mules and donkeys), died or were slaughtered due to suspected or confirmed AHS. One year after ending vaccination, December 1991, Portugal was declared free of AHS. Cost of eradication was US$1955513 (US$11.5/Portuguese equine).

African Horse Sickness↗

[The phylogenesis of equine teeth].

Phylogenetic research on the teeth of equidae beginning with dagger Hyracotherium ('Eohippus') will be summarized, subdivided into its components, analysed in a new way and finally interpreted according to several theories of evolution. In this context, specific cells (ameloblasts, odontoblasts, cementoblasts) are discussed and valued according to Preuss (1987) as active carriers of the evolution of teeth.

Animals↗

A blocking ELISA for detection of antibody to a subgroup-reactive epitope of African horsesickness viral protein 7 (VP7) using a novel gamma-irradiated antigen.

A novel gamma irradiated inactivated cell culture derived African horsesickness viral (AHSV) antigen was used in a blocking ELISA (B-ELISA) for detecting antibody to a subgroup-reactive epitope of AHSV. A monoclonal antibody (MAB), class IgM, against an epitope on African horsesickness (AHS) viral protein 7 (VP7) was developed in BALBc mice and used in the B-ELISA. The MAB, designated F9H, was blocked by 69 serums from equidae with antibody to AHS, but its binding activity was not appreciably affected by 301 serums that did not contain antibodies to AHS virus. An ELISA protocol using a blocking format is described.

African Horse Sickness↗

Estimation of the liveweight of working mules in Morocco from their body measurements.

During March 2001, 117 working mules, randomly chosen from four markets in Morocco, were weighed and a series of body measurements recorded; their age and body condition score were also recorded. Linear regression techniques were used to derive a 'best fit' equation for predicting liveweight from the other variables. For the working mules weighing between 131 and 391 kg the best prediction equation using two variables was: liveweight (kg) = -33 + 2.8 x heart girth (cm)+1.36 x length (cm). Other prediction equations available for estimating the liveweight of equidae were tested on the data, but in all cases they significantly overestimated the weights of the Moroccan mules.

Animals↗

Karyotypic relationships of horses and zebras: results of cross-species chromosome painting.

Complete sets of chromosome-specific painting probes, derived from flow-sorted chromosomes of human (HSA), Equus caballus (ECA) and Equus burchelli (EBU) were used to delineate conserved chromosomal segments between human and Equus burchelli, and among four equid species, E. przewalskii (EPR), E. caballus, E. burchelli and E. zebra hartmannae (EZH) by cross-species chromosome painting. Genome-wide comparative maps between these species have been established. Twenty-two human autosomal probes revealed 48 conserved segments in E. burchelli. The adjacent segment combinations HSA3/21, 7/16p, 16q/19q, 14/15, 12/22 and 4/8, presumed ancestral syntenies for all eutherian mammals, were also found conserved in E. burchelli. The comparative maps of equids allow for the unequivocal characterization of chromosomal rearrangements that differentiate the karyotypes of these equid species. The karyotypes of E. przewalskii and E. caballus differ by one Robertsonian translocation (ECA5 = EPR23 + EPR24); numerous Robertsonian translocations and tandem fusions and several inversions account for the karyotypic differences between the horses and zebras. Our results shed new light on the karyotypic evolution of Equidae.

Animals↗

The current challenges of dourine: difficulties in differentiating Trypanosoma equiperdum within the subgenus Trypanozoon.

During its 20th annual meeting in Paris in May 1999, the OIE (World organisation for animal health) Ad Hoc Group on Non-Tsetse Transmitted Animal Trypanosomoses expressed the following concerns about dourine: the discrepancies in some of the results of the complement fixation test (CFT), which is the only international diagnostic test officially recognised by the International Organisation for the Transportation of Equidae; the persistence of suspected cases of dourine in some Asian, European and African countries; the impossibility of differentiating Trypanosoma equiperdum from Trypanosoma evansi and of isolating new strains of T. equiperdum from clinical cases that have appeared in various parts of the world since 1982. In the light of these concerns, it was decided, in agreement with the Directorate of the Federal Veterinary Services of Russia in Moscow, to perform comparative trials on the value of CFT/dourine at the OIE Reference Laboratory for dourine in Moscow (The All-Russian Research Institute of Experimental Veterinary Medicine) using reagents (antigens and sera) from seven countries with extensive experience in the field of dourine diagnosis, namely, South Africa, France, Italy, Germany, Russia, the United States of America and the People's Republic of China. It is thanks to the successful co-operation of these countries that the trials were made possible. Results showed an overall concordance and were submitted for consideration to the OIE Biological Standards Commission, the commission which is in charge of the Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. These trials serve as a starting point for further study, particularly in the following areas: the isolation of new strains of T. equiperdum from clinical dourine cases; the identification of specific markers for T. equiperdum which would make it possible to differentiate it from among the other species within the subgenus Trypanozoon; the experimental infection of horses with newly isolated T. equiperdum strains to compare their pathogenicity with those currently used in national diagnostic laboratories and with that of T. evansi; phylogenetic studies; the proposal and validation of new, internationally recognised diagnostic test(s) for dourine.

Animals↗

Control and eradication of African horse sickness with vaccine.

African horse sickness (AHS) is an infectious but no-contagious viral disease of equidae with high mortality in horses. The disease is caused by an arthropod-borne double-stranded RNA virus within the genus Orbivirus of the family Reoviridae transmitted by at least two species of Culicoides. Nine different serotypes have been described. The nine serotypes of AHS have been described in eastern and southern Africa. Only AHS serotypes 9 and 4 have been found in West Africa from where they occasionally spread into countries surrounding the Mediterranean. Examples of outbreaks that have occurred outside Africa are: in the Middle East (1959-1963), in Spain (serotype 9, 1966, serotype 4, 1987-1990), and in Portugal (serotype 4, 1989) and Morocco (serotype 4, 1989-1991). Laboratory diagnosis of AHS is essential. Although the clinical signs and lesions are characteristic, they can be confused with those of other diseases. Several techniques have been adapted for the detection of RNA segments, antibodies and antigen. Two types of vaccines have been described for AHS virus. Attenuated live vaccines (monovalent and polyvalent) for use in horses, mules and donkeys, are currently available, as well as a monovalent, serotype 4, inactivated vaccine, produced commercially but no longer available. New vaccines, including a subunit vaccine, have been evaluated experimentally. In this paper a review of the last AHS outbreaks in Spain, occurring during 1987-1990, and affecting the central and south part of the country, is presented. The role that vaccination played for the control and eradication of the disease, as well as other aspects such as climatological conditions, number of vectors and horse management, are also presented and evaluated.

African Horse Sickness↗

Rapid evolution of horse satellite DNA.

The major satellite of the horse genome consists of about 1 million copies of a 221-bp tandem repeat unit. By fluorescence in situ hybridization it has been localized in the centromeres of 58 of the 64 horse chromosomes. The donkey genome contains a similar but not identical satellite. Strikingly, the equine repeat did not hybridize to DNA of the Grevy zebra, despite the divergence of the horse and zebra only 3 to 5 million years ago and the ability of these species to crossbreed. The evolution of satellite DNA in the Equidae is more rapid than that in other mammalian families, which may be explained by their rapid karyotypic evolution.

Animals↗

Spinal hyperostosis in comparative pathology. A useful approach to the concept.

Spinal hyperostosis, an anatomical and radiological concept primarily described in man, is characterized by enthesopathic bony overgrowth on vertebral bodies in the form of spurs or intervertebral bridges. It can also be part of a more diffuse enthesopathic condition, including the appendicular skeleton. These changes are distinct from those of osteoarthrosis. Similar changes can be observed in all kinds of mammals, independent of their type of locomotion (bipodic, quadrumanous, quadrupedic, or aquatic). An anatomical and radiological study is presented of six cases (with histological examination of two dogs and one horse, and observation of macerated specimens of one horse, one equida, and one whale). Comparative pathology allows for a better understanding of the concept of hyperostosis and, even more clearly than in man, this can be distinguished from that of discovertebral pathology although both conditions could be associated in older subjects. The knowledge of spinal hyperostosis in veterinary medicine could--as in man--be of interest to differentiate between this process and other features of vertebral pathology.

Animals↗

The equine enteric nervous system--neuron characterization and distribution in adults and juveniles.

A study of myenteric and submucosal plexuses was undertaken in the jejunum and ileum of horses and ponies in which no clinical or pathological evidence of intestinal abnormality was apparent. Complete transverse sections of the intestine, stained by a modified haematoxylin and eosin method, were examined using up to 20 sequential sections per animal. Information was gathered from adult, juvenile and fetal equidae. In adults, the longitudinal muscle layers were thinner than the circular muscle layers and the ileum had thicker layers compared to the jejunum. In adults, the submucosal plexus had more neurons per section than the myenteric plexus by mean ratios of 1:3 in the jejunum and 1:1.9 in the ileum. In juveniles, the ratios were respectively 1:1.8 and 1:1.5 and in the fetus 1:2.5 and 1:1.3. The three-dimensional distribution of neurons in both plexuses varied from animal to animal and no consistent pattern was observed. Groups of neurons contained between one and 42 cells per section examined and their length in a cranio-caudal direction varied from 10 to over 100 microns. There were few statistical differences observed between the cranial, middle and caudal portions of either the jejunum or the ileum when neuron groups or neuron numbers per section were examined in 10 adult animals.

Aging↗

Molecular timing of primate divergences as estimated by two nonprimate calibration points.

The complete mitochondrial DNA (mtDNA) molecule of the hamadryas baboon, Papio hamadryas, was sequenced and included in a molecular analysis of 24 complete mammalian mtDNAs. The particular aim of the study was to time the divergence between Cercopithecoidea and Hominoidea. That divergence, set at 30 million years before present (MYBP) was a fundamental reference for the original proposal of recent hominoid divergences, according to which the split among gorilla, chimpanzee, and Homo took place 5 MYBP. In the present study the validity of the postulated 30 MYBP dating of the Cercopithecoidea/Hominoidea divergence was examined by applying two independent nonprimate molecular references, the divergence between artiodactyls and cetaceans set at 60 MYBP and that between Equidae and Rhinocerotidae set at 50 MYBP. After calibration for differences in evolutionary rates, application of the two references suggested that the Cercopithecoidea/Hominoidea divergence took place >50 MYBP. Consistent with the marked shift in the dating of the Cercopithecoidea/Hominoidea split, all hominoid divergences receive a much earlier dating. Thus the estimated date of the divergence between Pan (chimpanzee) and Homo is 10-13 MYBP and that between Gorilla and the Pan/Homo linage approximately 17 MYBP. The same datings were obtained in an analysis of clocklike evolving genes. The findings show that recalculation is necessary of all molecular datings based directly or indirectly on a Cercopithecoidea/Hominoidea split 30 MYBP.

Animals↗

Assessment of pregnancy in Kiang mares (Equus hemionus holdereri ) using estrogen determination in feces.

Analysis of fecal estrogens was used to diagnose pregnancy in 6 Kiang mares (Equus hemionus holdereri ) that were kept at Tierpark Berlin. Three extraction methods were compared and were followed by an established RIA for total estrogen. Extraction of desiccated feces with chloroform/n-hexane and KOH, with and without enzyme hydrolysis showed better results than extraction with diethylether without hydrolysis. Pregnancy was confirmed by observation of foaling in 2 mares that showed estrogen concentrations between 800 and 1800 ng/g and in 1 mare that showed widely fluctuating values between 500 and 1300 ng/g of feces. Two mares with estrogen concentrations below 500 ng/g were not seen to foal. The method using chloroform/n-hexane and KOH without enzyme hydrolysis seems practical for non-invasive evaluation of the endocrine status in this endangered Equidae species.

Journal Article↗

Calcium excretion in feces of ungulates.

1. Fecal excretion of calcium was examined in 122 individual ungulates representing 7 species of Equidae, 3 species of Tapiridae, 3 species of Rhinocerotidae, 2 species of Elephantidae, 2 species of Hippopotamidae, 12 species of Bovidae, 2 species of Cervidae, 3 species of Camellidae and 1 species of Giraffidae. 2. Animals were fed timothy hay, a low calcium diet or alfalfa hay, a high calcium diet. 3. In a few cases oat straw or prairie hay was used instead of timothy hay. 4. Samples of feces were obtained from individuals daily for 4 days following a 20 day dietary equilibration period. 5. Feces of equids, tapirs, rhinoceros and elephants had a lower calcium concentration and a lower Ca/P ratio than feces of ruminants when the animals were fed diets of equivalent calcium content. 6. The findings suggest that the non-ruminant ungulate equids, tapirs, rhinoceros and elephants absorb a larger proportion of dietary calcium than ruminants do.

Animals↗

Equine parafilariosis in Iran.

From March to September 1991, horses (n = 1567), donkeys (n = 112) and mules (n = 96) were examined for equine parafilariosis by random sampling from different areas of Iran. The clinical signs of equine parafilariosis were observed in 136 cases of two northern areas (1.4% and 41.3% infection rate) and one northeastern area (6.3% infection rate). Most of the infected cases were confirmed by laboratory findings. All infected cases were followed up to establish the epidemiological features. The results suggest that there is one enzootic area in Iran, the Caspian coast and Persian Turkman steppes. An annual period of appearance of clinical signs coinciding with warm months was observed in all infected areas. No clinical sign was observed in cases less than 2 or more than 15 years of age. The highest infection rates (41.1-73.3%) were observed in equidae 4-9 years of age. The enzootic area encompasses the Caspian littoral, steppes and forest steppes and has an altitude of up to 1500 m. It has a temperature-wet climate. The treatment of 15 infected cases with ivermectin subcutaneously at 0.2 mg kg-1, was 100% effective with only one injection.

Aging↗

African horse sickness in Spain.

The aetiology, pathogenesis and epizootiology of African horse sickness (AHS) are reviewed with special reference to recent outbreaks in the Iberian peninsula. AHS is a highly fatal insect-borne viral disease of Equidae. It is caused by an Orbivirus (family Reoviridae) and nine serotypes are recognised. Outbreaks occurred in central Spain in 1987 and in southern regions of the Iberian peninsula in 1988, 1989 and 1990. All were associated with serotype 4 of the virus, whereas other occurrences of AHS outside Africa have all been caused by serotype 9. The clinical picture in the outbreaks was mainly of the acute (pulmonary) form except in 1988 when the subacute (cardiac) form of disease predominated. Several hundred horses died or were destroyed as a result of the outbreaks. Further spread was contained by a combination of slaughter of sick animals, movement controls, and vaccination which was extended over an increasingly wide area in successive years. The 1987 outbreak is believed to be associated with infected zebras imported from Africa. Possible explanations for the recurrence of disease in Spain in successive years are considered to include (a) the climatic conditions in Southern Spain, which could permit continuous vector activity, (b) the relative clinical resistance of mules and donkeys, which may permit subclinical circulation of the virus, (c) incomplete population immunity among horses due to possible gaps in the vaccination strategy.

African Horse Sickness↗