Subchromosomal karyotype evolution in Equidae.

Equidae is a small family which comprises horses, African and Asiatic asses, and zebras. Despite equids having diverged quite recently, their karyotypes underwent rapid evolution which resulted in extensive differences among chromosome complements in respective species. Comparative mapping using whole-chromosome painting probes delineated genome-wide chromosome homologies among extant equids, enabling us to trace chromosome rearrangements that occurred during evolution. In the present study, we performed subchromosomal comparative mapping among seven Equidae species, representing the whole family. Region-specific painting and bacterial artificial chromosome probes were used to determine the orientation of evolutionarily conserved segments with respect to centromere positions. This allowed assessment of the configuration of all fusions occurring during the evolution of Equidae, as well as revealing discrepancies in centromere location caused by centromere repositioning or inversions. Our results indicate that the prevailing type of fusion in Equidae is centric fusion. Tandem fusions of the type telomere-telomere occur almost exclusively in the karyotype of Hartmann's zebra and are characteristic of this species' evolution. We revealed inversions in segments homologous to horse chromosomes 3p/10p and 13 in zebras and confirmed inversions in segments 4/31 in African ass, 7 in horse and 8p/20 in zebras. Furthermore, our mapping results suggested that centromere repositioning events occurred in segments homologous to horse chromosomes 7, 8q, 10p and 19 in the African ass and an element homologous to horse chromosome 16 in Asiatic asses. Centromere repositioning in chromosome 1 resulted in three different chromosome types occurring in extant species. Heterozygosity of the centromere position of this chromosome was observed in the kiang. Other subtle changes in centromere position were described in several evolutionary conserved chromosomal segments, suggesting that tiny centromere repositioning or pericentric inversions are quite frequent in zebras and asses.

A comparative study of meiotic recombination in cattle (Bos taurus) and three wildebeest species (Connochaetes gnou, C. taurinus taurinus and C. t. albojubatus).

The karyotypic evolution in the family Bovidae is based on centric fusions of ancestral acrocentric chromosomes. Here, the frequency and distribution of meiotic recombination was analyzed in pachytene spermatocytes from Bos taurus (2n = 60) and 3 wildebeest species (Connochaetes gnou, C. taurinus taurinus and C. t. albojubatus) (2n = 58) using immunofluorescence and fluorescence in situ hybridization. Significant differences in mean numbers of recombination events per cell were observed between B. taurus and members of the genus Connochaetes (47.2 vs. 43.7, p < 0.001). The number of MLH1 foci was significantly correlated with the length of the autosomal synaptonemal complexes. The average interfocus distance was influenced by interference. The male recombination maps of bovine chromosomes 2 and 25 and of their fused homologues in wildebeests were constructed. A significant reduction of recombination in the fused chromosome BTA25 was observed in wildebeests (p = 0.005). This was probably caused by interference acting across the centromere, which was significantly stronger than the intra-arm interference. This comparative meiotic study showed significant differences among the species from the family Bovidae with the same fundamental number of autosomal arms (FNa = 29) which differ by a single centric fusion.

Molecular insights into X;BTA5 chromosome rearrangements in the tribe Antilopini (Bovidae).

For a clade that includes Antilope, Gazella,Nanger and Eudorcas (Antilopinae), X;BTA5 translocation is a synapomorphy. Using a combination of fluorescence in situ hybridization (FISH) probes and polymerase chain reaction techniques, we provide (i) the first insight into the X;BTA5 architecture which differs in the species under study: Antilope cervicapra (genus Antilope), Gazella leptoceros (genus Gazella) and Nanger dama ruficollis (genus Nanger), (ii) determination of interstitial satellite DNA at the X;BTA5 junctions, and (iii) determination of repetitive sequences occupying constitutive heterochromatin of Xp arms in the studied species. The distribution of 2 repetitive DNA families in the centromeric regions of all chromosomes has been investigated by FISH with probes representing satellite I and satellite II DNA in all studied species. In this context, we discuss a markedly smaller centromere in the BTA5 (Y2) unfused chromosomes in males in the XY1Y2 determining system in comparison with other acrocentrics. An analysis of karyotypic data described in current published studies revealed a disparity with the data determined by FISH. In this report, we document chromosomal fusions in the 3 species mentioned resulting from FISH with painting probes prepared from cattle (Bos taurus). The number and chromosomal location of nucleolus organizer regions were determined by FISH. In the present study, we emphasize the importance of chromosomal rearrangement verification, particularly, if they are used for phylogenetic analysis.

Cytotypes of Kirk's dik-dik (Madoqua kirkii,Bovidae) show multiple tandem fusions.

Madoqua kirkii, a miniature African antelope, is noted for extensive chromosomal variation that has been categorized in four distinct cytotypes (A-D). In this investigation, we analyzed the A cytotype (2n = 46, FN = 48) using a suite of molecular cytogenetic approaches that entailed (i) whole chromosome and subchromosomal painting by fluorescence in situ hybridization (FISH), (ii) the study of Madoqua centromeric-specific DNA derived from pooled DNA obtained from the centromeric regions of the acrocentric chromosomes, and (iii) DNA from the telomere:centromere junctions of tandemly fused chromosomes. DNA from these sources was used to probe for the persistence of interstitial satellite DNA and residual centromeric sequences in the tandem and centric fusion junctions by PCR and FISH. The analyses show centromeric sequences at two of the six tandem fusion junctions. These data, and those of hybrid specimens (A × B cytotypes) in conjunction with published information permitted an interpretation of the probable sequence of chromosomal rearrangements among the M. kirkii cytotypes. We discuss the findings in the context of chromosomal evolution in these antelopes, and the implications that these hold for ex-situ breeding programs of the species.

Different fusion configurations of evolutionarily conserved segments in karyotypes of Potamochoerus porcus and Phacochoerus africanus.

The karyotype of the red river hog Potamochoerus porcus (2n = 34) differs from that of the domestic pig by the presence of 2 fusion chromosomes homologous to pig chromosomes 13/16 and 15/17. Moreover, chromosomes corresponding to pig chromosomes 13/16 and 1 are both acrocentric. Hybridization with region-specific painting probes confirmed tandem fusion of pig chromosomes 13 and 16, and a pericentric inversion of the pig chromosome 1p equivalent in P. porcus. The chromosome complement of the wart hog Phacochoerus africanus (2n = 34) differs from the pig karyotype in 2 centric fusions, 13/16 and 15/17. Karyotypic relationships among different Suidae species are discussed in the article. Besides fusions 13/16 and 15/17, which are common to several suids, another fusion of pig chromosomes 14 and 18 is suggested to exist in the karyotype of Sus cebifrons.

Karyotype, centric fusion polymorphism and chromosomal aberrations in captive-born mountain reedbuck (Redunca fulvorufula).

Chromosomes of fourteen captive-born mountain reedbucks (Redunca fulvorufula) have been investigated. The diploid chromosome number was 2n = 56 (FN = 60). The mountain reedbuck karyotype consists of 26 acrocentric and two biarmed chromosome pairs resulting from two centric fusions involving chromosomes 2 and 25, and 6 and 10, respectively. In some animals, 57 chromosomes were detected. Variation in the diploid number was found to be due to polymorphism for the centric fusion 6;10. Both X and Y chromosomes are large and acrocentric. The entire Y chromosome and the proximal part of the X chromosome consist of heterochromatin. The chromosomes X, 9 and 14 appeared to be of caprine type. Chromosome aberrations have been detected in two of the 14 animals investigated. A de novo formed Robertsonian translocation rob(6;13) was found in one female heterozygous for the fusion 6;10. CBG-banding revealed one block of centromeric heterochromatin in the de novo formed translocation rob(6;13) and also in the evolutionarily fixed centric fusions 6;10 and 2;25. One examined male homozygous for fusion 6;10, had a mosaic 56,XY/57,XYY karyotype, with 11% of analyzed cells containing two Y chromosomes. The findings were confirmed by cross-species fluorescence in situ hybridization (FISH) with bovine (Bos taurus L.) chromosome painting probes. The study demonstrates the relevance of cytogenetic screening in captive animals from zoological gardens.

Faecal progesterone metabolite analysis for non-invasive monitoring of reproductive function in the white rhinoceros (Ceratotherium simum).

The two subspecies of white rhinoceros, southern (Ceratotherium simum simum) and northern (Ceratotherium simum cottoni), breed poorly in captivity, and estimates of oestrous cycle length vary considerably (range, 25-90 days). To characterise reproductive patterns, faecal samples were collected 2-3 times/week for up to 56 months from non-pregnant animals (n=21) of both subspecies. Immununoreactive pregnanes containing a 20-oxo-group (20-oxo-P) were analysed in a group-specific enzyme immunoassay using an antibody against 5alpha-pregnane-3beta-ol-20-one 3HS:BSA. Reproductive patterns were highly variable among and within individual animals. However, rhinoceroses could be classified into four major categories on the basis of oestrous cycle length and luteal phase 20-oxo-P concentrations: (1) regular oestrous cycles of 10 weeks duration and > 800 ng/g (n=2 animals); (2) oestrous cycles between 4-10 weeks and 250-750 ng/g (n=6); (3) no apparent cycle regularity, but luteal activity indicated by 20-oxo-P concentrations of 100-200 ng/g (n=6); (4) no apparent luteal activity as indicated by 20-oxo-P of < 100 ng/g (n=7). In two attempts to induce ovarian activity, chlormadinone acetate was fed daily to one animal for 35 and 45 days, respectively. Each treatment was followed by a subsequent hCG injection which resulted in luteal phases of 17 and 18 days, respectively, beginning about 10 days after hCG. Concentration of faecal 20-oxo-P in one pregnant animal during the 4th and 5th month of gestation were markedly higher than those observed during the luteal phase of the cycle. In conclusion, two thirds of white rhinoceroses in this study had erratic or missing luteal activity, whereas variable cycles of 4-10 weeks in length were evident in six females, and regular oestrous cycles of 10 weeks in length were found in two animals.

Assessment of the risk of phosphorus loading due to resuspended sediment.

Resuspension is a multiphase phenomenon where suspended solids encounter water layers differing in physico-chemical properties that affect the reactions of phosphorus (P). The role of resuspended sediment as a sink or source of dissolved P was determined in a laboratory study of P desorption-sorption equilibria. Gradual mixing was simulated using decreasing solid concentrations and varying environmental conditions (pH, redox, ionic strength). To describe the P exchange when the particles encounter dissimilar water layers, the extent of P sorption to or desorption from solids was expressed as a function of P concentration in the bath solutions. The equilibrium phosphorus concentration (EPC), at which there is no net P release from or retention to the particles, proved to be a suitable parameter for assessment of P load risk. Under oxic conditions at pH 7, commonly prevailing in lakes, the EPC values ranged from 11 to 27 microg P L(-1). The larger the water volume the suspended material was mixed with, the higher the P concentration, allowing desorption to occur. As for chemical factors affecting P mobilization, EPC followed the order: pH 7 < pH 7 anoxic < pH 9. A separate extraction experiment revealed that elevated pH enhanced P mobilization more as the concentration of solids decresed. The results demonstrate that high pH (a common characteristic in eutrophic lakes during summer), when linked with intensive resuspension, may markedly increase the internal P loading risk. As for the risk assessment, the quantification of the internal P loading would be improved by isotherm studies combined with field observations.

Phosphorus mobilization from various sediment pools in response to increased pH and silicate concentration.

Phosphorus (P) release from sediment particles to the interstitial water has been studied extensively, but the contribution of different inorganic P pools in sediment under differing environmental conditions is not fully understood. This study was undertaken to get more detailed information about the chemical mobilization mechanisms. Phosphorus mobilization from reserves bound by Al, Fe, and Ca compounds in response to increased pH and to inorganic silicon (Si) enrichments was investigated using a sequential fractionation analysis and an isotope-labeling technique. The aerobic sediment of Lake Vesijärvi had a high P retention capacity, and Fe-bound P was the largest inorganic P pool as well as the main source of released P. High Si addition (47 mg Si L-1 sediment) released more P to the interstitial water than did the elevation of pH from 6.6 to 9.5, since Si lowered the resorption of released P onto hydrated Al oxides. This finding reveals that P equilibrium between Fe-bound and Al-bound P in sediments regulates P net mobilization to the interstitial water under aerobic conditions. Furthermore, elevated pH combined with high Si enrichment had a positive synergistic effect, resulting in the most substantial P mobilization. This synergism may cause a self-fueled increase in the internal loading of P. It accentuates the effect of diatom sedimentation on P fluxes in eutrophic lakes with high pH and may favor the appearance of bloom-forming cyanobacteria.

Pathological process induced by Demodex sp. (Acari: Demodicidae) in the skin of eland, Taurotragus oryx (Pallas).

Taxonomic characters of Demodex sp. (near to D. kutzeri Bukva, 1987) are described and discussed. Pronounced, disperse, chronic, multinodular folliculitis induced by this hair follicle inhabitant in the skin of eland, Taurotragus oryx, is described at the histological level.

[Clinical experience and comparison of ketamine-medetomidine with ketamine-xylazine anesthesia in the African wild dog (Lycaon pictus) in captivity]. / Klinické zkusenosti a srovnání ketamín-medetomidin a ketamín-xylazín anestézie psa hyenového (Lycaon pictus) v zajetí.

The effects of two mixtures, ketamine-xylazine and ketamine-medetomidine, were compared in anesthesia of African wild dog (Lycaon pictus) in the Zoo park at Dvur Králové; the effects of these combinations were also investigated on the triad values and on the basic hematological (red blood counts, hematocrit, hemoglobin content, derived parameters MCM, MCHC, MCV, white blood counts, differential blood counting) and biochemical (total proteins, glucose, creatinine, urea, cholesterol, magnesium, calcium, phosphorus, chlorides, sodium, potassium, alkaline phosphatase, gamma-glutamyl transferase, alanine transferase, aspartate transaminase) parameters of blood and blood serum in anesthesia. Tab. I shows the reasons for anesthesia in 68 individuals of African wild dog in the years 1980-1990. As for both mixtures, application of drugs with a blowpipe was used. In the anesthetized animals, the outset of ataxia, laying down and the outset of sleep were followed (Tab. II), as well as the time of wakening up without and with antidote administration (Tabs. IV and V). The outset of ataxia was fast 1.5 +/- 0.6 min) in the ketamine-xylazine mixture administered at doses of 5.07 +/- 1.16 mg/kg ketamine and 2.11 +/- 0.53 mg/kg xylazine, similarly like lying down (3.2 +/- 1.0 min) and loosing sensation (6.3 +/- 1.6 min). At the start of the drug action, vomiting was often observed, and sometimes in the first ten minutes after drug harpooning short clonic convulsions of the limbs or the whole body occurred. In further course, immobilization and anesthesia were complete and satisfactory in all cases. The first reactions to outer stimuli during wakening up without antidote administration were observed in 135 +/- 11.9 minutes while the animals stood up in 210 +/- 44.5 minutes after drug harpooning (Tab. IV). When the nonspecific antidote xylazine-yohimbine was used, the first reactions after i.m. instillation at a dose of 0.31 +/- 0.02 mg/kg appeared in 20.3 +/- 0.57 min, after i.v. instillation at a dose of 0.11 +/- 0.17 it was in 9.3 +/- 4.16 min after antidote administration. The animals stood up and started walking in 53.3 +/- 24.3 min after i.v. instillation (Tab. V). No significant changes (Tab. III) were observed in the values of temperature, breathing rate and pulse rate when the variations of triad values were investigated in five individuals within the first 30 minutes (up to 10 min, 20 and 30 min) after drug harpooning.(ABSTRACT TRUNCATED AT 400 WORDS)

Demonstration of parvovirus in diarrhoeic African cheetahs (Acinonyx jubatus jubatus Schreber, 1775).

Parvovirus was demonstrated in the intestinal content of diarrhoeic African cheetahs by electron microscopy. The virus was isolated in a feline kidney cell line inoculated with a filtrate of the intestinal content. Its growth characteristics, cytopathic effect, agglutination of porcine erythrocytes, structure, and results of immunoelectron microscopic examination were indistinguishable from those of feline panleukopenia virus.

Karyotypic relationships among Equus grevyi, Equus burchelli and domestic horse defined using horse chromosome arm-specific probes.

Using laser microdissection we prepared a set of horse chromosome arm-specific probes. Most of the probes were generated from horse chromosomes, some of them were derived from Equus zebra hartmannae. The set of probes were hybridized onto E. grevyi chromosomes in order to establish a genome-wide chromosomal correspondence between this zebra and horse. The use of arm-specific probes provided us with more information on the mutual arrangement of the genomes than we could obtain by means of whole-chromosome paints generated by flow sorting, even if we used reciprocal painting with probe sets from both species. By comparison of our results and results of comparative mapping in E. burchelli, we also established the chromosomal correspondence between E. grevyi and E. burchelli, providing evidence for a very close karyotypic relationship between these two zebra species. Establishment of the comparative map for E. grevyi contributes to the knowledge of the karyotypic phylogeny in the Equidae family.

Serum chemistry profiles for Lechwe waterbucks (Kobus leche): variations with age and sex.

1. Over an 8-year period, 19 biochemical parameters have been determined at various ages in the blood serum of 92 clinically healthy Lechwe waterbucks (Kobus leche), 33 males and 59 females. 2. Significant differences have been noted with age. In neonates, the lowest values of total proteins, glucose, creatinine, urea, AST, ALT and iron have been noted; the highest ones have been seen for cholesterol, alkaline phosphatase, calcium and phosphorus. 3. With regard to sex, raised values of glucose, urea, alkaline phosphatase and ALT, and lowered values of cholesterol, have been noted in juvenile females compared with males of the same age. 4. In adult females, higher levels of urea and cholesterol and lower levels of glucose, triglycerides and natrium have been recorded compared with males. 5. With sex and age, no significant changes have been found in the levels of GGT, magnesium, chlorides and copper. 6. Our findings are discussed with those abstracted from the literature for related species.

Time-dependent influence of some sedating agents on basic haematological values in various artio- and perissodactylids.

Basic haematological values in 32 animals of five species were estimated after administration of sedating agents. In all species under investigation, a time-dependent decrease of erythrocyte counts, haematocrit values and haemoglobin content was noted during the first 30 min after sedation, for the following 30 min the lowered values remained essentially without any change in zebras. Derived parameters, mean corpuscular haemoglobin, mean corpuscular Hb concentration and mean corpuscular volume did not change during the period of observation. Only insignificant changes in leukocyte count and in the proportion of lymphocytes and neutrophiles were registered. The shortest possible time between sedation and blood sampling is recommended to minimalize a distortion especially in the red blood picture.

Basic haematological values in the Cameroon goat (Capra hircus).

1. Basic haematological values in 165 Cameroon goats (Capra hircus) are reported. 2. The erythrocyte count, mean 14.36 x 10(12) l-1, ranged from 8.24 to 24.7 x 10(12) l-1; the haematocrit values, mean 0.304, varied from 0.20 to 0.38; the haemoglobin content, mean 113.4 g/l, was in the range from 83.0 to 143.0 g/l and the leukocyte count, mean 13.67 x 10(9) l-1, had lowest and highest values between 5.4 and 24.5 x 10(9) l-1. 3. Comparing these blood constituents in 47 male and 118 female Cameroon goats we demonstrated statistically significant lower values of the haematocrit and haemoglobin content and a statistically significant higher proportion of lymphocytes in the female animals. 4. In 16 pregnant and 30 non-pregnant female Cameroon goats, all animals older than 3 years, no statistically significant differences of the red blood picture were noted. 5. Also in three groups, assorted according to age, no significant changes in basic haematological parameters were seen. 6. During one year follow-up of some haematological parameters, statistically significant seasonal changes were found. 7. All presented data are compared with values abstracted from the literature and discussed.

Basic haematological values in carnivores--II. The Felidae.

1. Basic haematological values in 34 animals of eight carnivorous species are reported. 2. In four Northern lynxs (Lynx lynx lynx), two male and two female animals, the mean values are given: erythrocyte counts 8.51 X 10(12)/l, haematocrit 0.392/l, haemoglobin content 148.0 g/l and leukocyte count 7.92 X 10(9)/l. 3. In six male pumas (Puma concolor missolensis) the mean values estimated are: erythrocyte count 9.35 X 10(12)/l, haematocrit 0.43/l, haemoglobin content 163.9 g/l and leukocyte count 7.73 X 10(9)/l. Individual values in one female puma are also given. 4. In six jaguars (Panthera onca), three male and three female animals, the mean values are given: erythrocyte count 8.27 X 10(12)/l, haematocrit 0.37/l, haemoglobin content 137.1 g/l and leukocyte count 15.15 X 10(9)/l. 5. Only individual values are reported in one clouded leopard (Neofelis nebulosa), in one leopard (Panthera pardus saxicolor), in one Corbett's tiger (Panthera tigris Corbetti) and in one Altaic tiger (Panthera tigris Altaica). 6. In four lions (Panthera leo leo), two male and two female animals, the mean estimated values are: erythrocyte count 10.14 X 10(12)/l, haematocrit 0.462/l, haemoglobin content 159.0 g/l and leukocyte count 11.05 X 10(9)/l. In six female cheetahs (Acinonox jubatus jubatus) the mean values estimated are: erythrocyte count 7.86 X 10(12)/l, haematocrit 0.373/l, haemoglobin content 142.8 g/l and leukocyte count 8.65 X 10(9)/l. For three male cheetahs only individual values are reported. 8. All results achieved are compared with those abstracted from the literature and discussed.

Basic haematological values in carnivores--I. The Canidae, the Hyaenidae and the Ursidae.

Basic haematological values for 32 animals of four carnivore species are reported. In six adult wolves (Canis lupus) the mean values estimated for the erythrocyte count 7.48 X 10(12)/l, haematocrit 0.465/l, haemoglobin 172.0 g/l and leukocyte counts 7.33 X 10(9)/l are given. For five young wolves, these parameters were markedly decreased, only the white cell count was raised. In 14 hunting dogs (Lycaon pictus) the mean values estimated are: erythrocyte count 9.15 x 10(12)/l haematocrit 0.435/l, haemoglobin 179.2 g/l and leukocytes 12.95 X 10(9)/l. In six striped hyaenas (Hyaena hyaena) the mean estimated values are: erythrocyte count 8.11 X 10(12)/l, haemoatocrit 0.445/l, haemoglobin 178.0 g/l and leukocytes 13.95 X 10(9)/l. Only individual values for the reported parameters are given in the Asiatic black bear (Selenarctos thibetanus). All results are compared with values derived from the literature for animals under investigation and for the domestic dog (Canis familiaris).

Basic haematological values in the African buffalo (Syncerus caffer caffer) and in the red buffalo (Syncerus caffer nanus).

Basic haematological values in 13 African buffaloes (Syncerus caffer caffer), 11 cows and 2 bulls, and in 2 red buffaloes (Syncerus caffer nanus), 1 bull and 1 cow, are reported. In the African buffalo, the erythrocyte count--mean 9.76 X 10(12) 1(-1)--ranged from 6.72 to 12.42 X 10(12) 1(-1), the haematocrit values--mean 0.391--varied from 0.32 to 0.47 and the haemoglobin content--mean 148.17--was in the range from 122.1 to 172.5 g 1(-1). For the red buffalo only individual values are given. In the African buffaloes, the leukocyte counts--mean 7.39 X 10(9) 1(-1)--ranging from 5.1 to 12.4 X 10(9) 1(-1) were slightly increased compared with man. For the red buffalo only individual values are reported. These values are compared with data abstracted from the literature for related buffalo species. The age-dependent decrease of erythrocyte counts and the age-dependent rise of the eosinophile proportion in the differential count are discussed.