Development of the skeleton in mouse fetuses of both sexes between 14.5 and 18.5 days post coitum

Ossification during embryogenesis of female and male mouse fetuses between 14.5 and 18.5 days post coitum (d.p.c.) was investigated with two methods. The Alizarin-red based method allows differential staining of cartilage and bone tissue on whole animal, however standard histochemical methods allow detailed identification of cells involved in ossification process. There were minor differences in the time of occurrence of some ossification centers between male and female mouse fetuses in a period of 14.5 - 18.5 d.p.c. At 14.5 d.p.c. ossification of female but not male mandible was observed and at 18.5 d.p.c. histochemistry demonstrated ossification in the phalanges in male but not in female.     

Hematology of equine fetuses with comparisons to their dams

The Hematologic values of 19 equine fetuses between 202 and 238 days gestation were compared with those of their dams. The red blood cell (RBC) count, hemoglobin concentration, hematocrit, and mean corpuscular hemoglobin concentration were significantly lower in fetal blood, while the mean corpuscular volume, mean corpuscular hemoglobin, and red cell distribution width were significantly higher. Mares had a significantly higher nucleated blood cell count than fetuses, and all nucleated cells were leukocytes (WBC). Most WBC in mare blood were segmented neutrophils and lymphocytes. In contrast, over one-half of the nucleated cells in fetal blood were nucleated RBC, and the majority of WBC in fetal blood were lymphocytes. Mares also had significantly higher plasma protein and fibrinogen concentrations than their fetuses. Mild macrocytosis and mild polychromasia were observed in most fetal blood samples, but not in blood samples from mares. All fetal blood contained reticulocytes, and most samples contained Heinz bodies and Howell-Jolly bodies. The results of this study will contribute to the development of hematologic reference values that may be useful in equine fetal research and, possibly, in the diagnosis of equine fetal disease.     

Pharmacokinetics of ceftiofur sodium in equine pregnancy

Eleven pregnant pony mares (D270‐326) were administered ceftiofur sodium intramuscularly at 2.2 mg/kg (n = 6) or 4.4 mg/kg (n = 5), once daily. Plasma was obtained prior to ceftiofur administration and at 0.5, 1, 2, 4, 8, 12, and 24 hr after administration. Eight pony mares were re‐enrolled in the study at least 3 days from expected foaling to ensure steady‐state concentrations of drug at the time of foaling. Mares were administered ceftiofur sodium (4.4 mg/kg, IM) daily until foaling. Parturition was induced using oxytocin 1 hr after ceftiofur sodium administration. Allantoic and amniotic fluid, plasma, and colostrum samples were collected at time of foaling. Serial foal plasma samples were obtained. Placental tissues were collected. Desfuroylceftiofur acetamide (DCA) concentrations were measured in samples by high‐performance liquid chromatography (HPLC). Mean (±SD) peak serum concentrations of DCA were 3.97 ± 0.50 μg/ml (low dose) and 7.45 ± 1.05 μg/ml (high dose). Terminal half‐life was significantly (p = .014) shorter after administration of the low dose (2.91 ± 0.59 hr) than after administration of the high dose (4.10 ± 0.72 hr). The mean serum concentration of DCA from mares at time of foaling was 7.96 ± 1.39 μg/ml. The mean DCA concentration in colostrum was 1.39 ± 0.70 μg/ml. DCA concentrations in allantoic fluid, amniotic fluid, placental tissues, and foal plasma were below the limit of quantification (<0.1 μg/ml) and below the minimum inhibitory concentration of ceftiofur against relevant pathogens. These results infer incomplete passage of DCA across fetal membranes after administration of ceftiofur sodium to normal pony mares.     

Molecular characterisation of the ORF68 region of equine herpesvirus-1 strains isolated from aborted fetuses in Hungary between 1977 and 2008

Equine herpesvirus-1 (EHV-1) can be classified into distinct groups by single nucleotide polymorphisms (SNPs) in their genomes. Only a few of these can be associated with a special attribute of the virus. Differences in the ORF30 region can determine the neuropathogenic potential, while by substitutions in the ORF68 region several strain groups can be made. In previous studies no connection was found between the neuropathogenic potential and the SNPs in ORF68, but the occurrence of members of distinct groups in different outbreaks can facilitate epidemiological investigations because the geographical distribution of a particular group is very often specific. The present study aimed at the molecular examination and grouping of 35 EHV-1 strains isolated from aborted equine fetuses in Hungary between 1977 and 2008. Genotyping was based on the comparison of nucleotide sequences of a polymorphic segment located in the ORF68 region, which had previously been found to be a useful tool for classification. After sequencing this region, the Hungarian EHV-1 isolates could be classified into seven groups. Only 23 of the 35 isolates belonged to the formerly described groups, while the SNPs of 12 isolates diverged, and four new groups could be set up. In addition, phylogenetic analysis was performed to compare the ORF68 sequences of the Hungarian strains with the sequences of isolates from Europe, America and Australia. The number of newly formed groups suggests that the further analysis of unknown EHV-1 isolates would involve the emergence of extended numbers of new groups, which can impair the usability of this grouping method.     

Laboratory methods of equine pregnancy diagnosis.

Rectal examination is a reliable method of diagnosing pregnancy in the mare. Also, test kits are available for the simple quick detection of pregnant mare serum gonadotrophin. Nevertheless there is a considerable demand by practitioners for an independent laboratory service in equine pregnancy diagnosis, particularly during the gestational phase when placental gonadotrophin is concentrated in the blood. An initial attempt to provide a service by means of the agar gel diffusion test was disappointing and alternatives were sought. The primary requirements for an ideal alternative technique were defined as: accuracy, sensitivity, applicability to the optimum request period, elimination of subjective interpretation and a minimal inconclusive rate. Additional considerations included cost, practicality and test duration. The various tests available are summarised and their published accuracies are discussed. It was decided to reverse the current trend from biological to immunological techniques and to modify the test by which Cole and Hart announced the discovery of PMSG in 1930. The utero-ovarian response in the immature female mouse was quantified simply to produce a numerical result. The reason for this is explained, the method is described and its accuracy is evaluated. The primary requirements, defined above, were achieved.     

马传染性贫血驴强毒gp90基因的克隆和序列分析

以EIAV驴强毒株D-AmRNA为模板，利用RT-PCR技术，扩增了约1．4kb的gp90基因。将其克隆后进行了测序。测序结果表明所扩增的1338个核苷酸片段含有完整的gp90基因全序列。核苷酸和氨基酸序列比较分析结果表明：D-A EIAV与国内分离株辽系强毒L株差异率仅有1．8％，而与国外毒株(克隆1369，WENVl7、WENVl6、PSPEIAVl9)核苷酸差异率在35．5％～37．2％之间；D-A株与国内分离株L株氨基酸水平差异率在2．9％，而与国外毒株氨基酸水平上的差异率在42．6％—46．0％；D-AEIAV有19个N-连接糖基化位点，L株、WENVl7和WENVl6是18个，克隆1369、WENVl6和WENVl7亲本毒株PSPEIAVl9是12个。     

Two related cases of cerebellar abnormality in equine fetuses associated with hydrops of fetal membranes

Hydrops allantois was diagnosed in two Haflinger mares with severe abdominal distension. Both mares were seven months pregnant. Abortion was induced with two injections of prostaglandin six hours apart followed by further manual dilation of the cervix and administration of oxytocin the next day. There were 90 and 95 litres of fluid, respectively, in the allantoic cavities which resembled extracellular fluid with regard to concentrations of urea, creatinine, sodium, potassium, calcium, magnesium, phosphate and chloride, but not total protein. Both fetuses had severe brain abnormalities which were diagnosed as cerebellar and cerebral hypoplasia associated with bilateral hydrocephalus internus and hydranencephaly and cerebellar aplasia, respectively. Both mares were pregnant by the same stallion, but a clear hereditary link was not found.     

Serum selenium levels in sheep from birth to 90 days of age]

The results of the study of the dynamics of selenium in the blood serum were compared in 31 piglets from birth to the age of 90 days. The analyzed selenium values in the blood serum immediately after birth (0.14--0.20 ppm) provide evidence of the intra-uterine placental passage of selenium. In the period of colostral and milk nutrition, the serum concentrations of selenium showed a statistically insignificant variation, followed by an abrupt drop of the level of the microelement in the weaning period (60th day). The individual values in this period ranged from 0.08 to 0.12 ppm. Towards the end of the study, the level of selenium in the blood serum of the piglets returned to its initial value; this indicates that although the critical period connected with weaning has a transient nature, it should be averted for preventing the occurrence of a selenium-deficit disease; this can be done by adding a supplement of 0.2% solution of sodium selenate or a combined preparation of selenium and vitamin E-Selevit.     

Evaluation of the long-term oral consequences of equine exodontia in 50 horses

The aims of this study were to objectively evaluate and quantify the process of post-extraction cheek teeth (CT) dental drift in horses, and to report on associated disorders of CT wear and long-term periodontal health. Fifty horses that had CT oral extraction because of apical infection were prospectively re-examined and a full oral examination, including measurements of some dental parameters, was performed.Narrowing of the extraction space was noted in all cases with complete closure occurring in 18% of horses. The rate of dental drift was calculated as 15.7% of extraction space/year (range 4–50%) and was not associated with the age at extraction (P = 0.78) or frequency of dental care since extraction (P = 0.48). There was a significant negative relationship between the rate of dental drift and the duration of time since extraction (P = 0.008). Overgrowths were present on the opposite CT row in 98% of horses, including opposite the extracted CT and on the Triadan 06s and 11s. No significant difference was noted in either the number of diastemata (P = 0.9) or periodontal disease score (P = 0.8) between the extraction and the contralateral cheek tooth rows.     

High prevalence of chlamydial (Chlamydophila psittaci) infection in fetal membranes of aborted equine fetuses

Seventy-seven cases of equine abortion from 49 Hungarian farms that occurred between 1998 and 2000 were investigated for the presence of chlamydiae by immunohistochemistry, PCR and/or MZN staining. Evidence of the presence of these bacteria was obtained in 64 cases (83.1%) from 41 (83.7%) different farms. Partial ompA gene sequencing of PCR products revealed that the agent was Chlamydophila psittaci. Based on the findings of microbial diagnosis, pathology and case history, chlamydial infection was considered to be the most likely cause of abortion in at least 11 (14.3%) cases. In the remaining 53 Chlamydophila-positive cases, either other bacterial and viral agents (n = 22 or 28.6%) as well as non-infectious factors (n = 14 or 18.2%) were identified as more probable primary causes of disease, or the role of chlamydiae remained unclear because lesions in fetuses and fetal membranes were absent (n = 17 or 22.1%). When chlamydial antigen was detected in aborted equine placental tissue using immunohistochemistry it was seen only in the chorionic epithelial cells, but not in other parts of the fetal membranes nor in any of the fetal tissues. In conclusion, chlamydial infection of the genital tract should be considered a possible factor in equine reproductive disorders.     

Plasma and blood volume in the calf from birth till 90 days of age.

Determinations of plasma volume were made of 9 clinically healthy Swedish Red and White calves from birth to 90 days of age by means of the isotop dilution technique. Commercially available ¹³¹I labelled human serum albumin was used. Calculation of the total blood volume was based on the plasma volume and packed cell volume.The plasma and blood volumes increased per kg body weight in average 17 and 14% respectively from directly after birth to 24 hrs. old. From 1 to 90 days of age the plasma and blood volume fell steadily per kg body weight. Plasma volume expressed as a percentage of body weight was 5.3% at birth, 6.5% at 1 day old, and 4.9% at 90 days old. Corresponding values for blood volume were 8.4, 9.3 and 7.0%.