[Cytogenetical and molecular analyses in a horse with SRY-negative sex reversal]. / Zytogenetische und molekulargenetische Abklärungen bei einem Pferd mit SRY-negativer Sex-Umkehr.

Numerical and structural aberrations of chromosomes may cause malformations, embryonal losses and reduced or missing fertility. In male horses the fertility is rather well controlled through their semen evaluation. For mares there are no mandatory regulations which specify their use in a breeding programme. Therefore, mares with chromosomal aberrations, which exclude reproduction success may be chosen for breeding. The present case describes a horse, offically registered as a female, which was presented for a breeding exam. On this occasion, small and inactive ovaries were diagnosed. Although the cytogenetical analysis excluded a numerical or gross structural chromosome aberration, the karyotype (64,XY) corresponded to the male sex. The diagnosis male to female sex-reversal (SRY-negative) excludes reproductive success for this horse. This case underlines the importance of a cytogenetic analysis for female horses, before they enter a breeding programme.

Interleukin-8 concentration and neutrophil chemotactic activity in bronchoalveolar lavage fluid of horses with chronic obstructive pulmonary disease following exposure to hay.

OBJECTIVE: To analyze effects of hay dust exposure on interleukin-8 (IL-8) concentration, percentage of neutrophils, and neutrophil chemotactic activity in bronchoalveolar lavage fluid (BALF) of horses with chronic obstructive pulmonary disease (COPD). ANIMALS: 16 healthy horses and 29 horses with COPD. PROCEDURE: IL-8 concentration, percentage of neutrophils, and neutrophil chemotactic activity in BALF were measured. Values were analyzed with respect to hay dust exposure. These variables were also measured in 5 asymptomatic horses with COPD after the induction of clinical signs by changing feed from silage to hay. RESULTS: L-8 concentrations and chemotactic activity in BALF were greater in horses with COPD, compared with healthy horses, and greater in horses with COPD exposed to hay dust, compared with nonexposed affected horses. An increase in IL-8 concentration accompanied by an increase in percentage of neutrophils in BALF and development of clinical signs of COPD were induced in asymptomatic horses with COPD by changing feed from silage to hay. CONCLUSIONS AND CLINICAL RELEVANCE: Exposure of horses with COPD to hay dust components resulted in an increase in IL-8 secretion at the bronchoalveolar surface. This chemokine may play a role in the pathogenesis of COPD, because it causes neutrophil accumulation in the bronchoalveolar space. Our results underscore the importance of eliminating dust sources for the treatment and prevention of COPD in horses.

Gastrointestinal and endocrine function during 'foal heat diarrhoea' in healthy foals.

Gastrointestinal function was assessed in normal foals before, during and after the onset of 'foal heat diarrhoea' by haematological, biochemical, microbiological and parasitological methods, and by a combined lactose tolerance-breath hydrogen excretion test. Warmblood mares (n=11) and their foals were used. The foals were assessed according to a standard protocol after birth and at regular intervals thereafter. All except one of the foals suffered from diarrhoea that started between day 8 and day 12 after birth, which lasted for 1-4 days. Rotavirus was detected in the faeces of five foals, but there was no association with the occurrence of diarrhoea. A significant increase in aldosterone concentration compared with that at days 6 and 18 was observed in five foals on the second day of diarrhoea. There were no differences between age groups in plasma glucose concentrations after lactose administration. Peak breath hydrogen excretion after lactose administration ranged from 17 to 232 p.p.m., and increased with age of foal. There was no decrease in lactose absorption or increase in breath hydrogen excretion in foals suffering from diarrhoea.

[Borna disease in Switzerland and in the principality of Liechtenstein]. / Die "Bornasche Krankheit" in der Schweiz und im Fürstentum Liechtenstein.

Borna disease (BD) is a rare immunopathological disorder of the central nervous system (CNS) caused by infection with Borna disease virus (BDV) and histologically characterized by mononuclear encephalomyelitis. BD primarily affects equines and sheep in well defined endemic areas of central Europe, but BDV infections have also been reported in other host species including humans, as well as in non endemic regions. In this paper recent data on the pathogenesis of BD are reviewed and the current situation in Switzerland and the Principality of Liechtenstein is summarized.

The role of neutrophil chemotactic cytokines in the pathogenesis of equine chronic obstructive pulmonary disease (COPD).

Chronic obstructive pulmonary disease (COPD) is defined as a chronic obstructive inflammatory disease affecting the small airways associated with hay dust exposure (Lowell, F.C., 1964. Observation on heaves. An asthma like syndrome in the horse, J. Allergy 35, 322-330). The disease corresponds histopathologically to a chronic bronchiolitis (Gerber, H., 1973. Chronic pulmonary disease in the horse, Equine Vet. J. 5, 26-33; Winder, N.C., Grünig, G., Hermann, M., Howald, B., von Fellenberg, R., 1989. Comparison of respiratory secretion cytology and pulmonary histology in horses, J. Vet. Med., A36, 32-38) and is mainly characterized by the presence of neutrophil granulocytes in the small bronchioles. Around 12-50% of all horses in Europe and the northern United States suffer from this disease (Mc Pherson, E.A., Lawson, G.H.K., Murphy, J.R., Nicholson, J.M., Fraser, J.A., Breeze, R.G., Pirie, H.M., 1978. Chronic obstructive pulmonary disease (COPD): Identification of affected horses, Eq. Vet. J. 10, 47-53; Larson, V.L., Busch, R.H., 1985. Equine tracheobronchial lavage: Comparison of lavage cytologic features in horses with chronic obstructive pulmonary disease, Am. J. Vet. Res., 46, 144-146; Bracher, V., von Fellenberg, R., Winder, N.C., Grünig, G., 1991. An investigation of the incidence of chronic obstructive pulmonary disease (COPD) in random populations of swiss horses, Equine Vet. J. 23, 136-141). The number of neutrophils in the bronchoalveolar lavage (BAL) and in tracheobronchial secretions (TBS) correlates with the severity of the disease. The present study is focused on the mechanisms which lead to the infiltration of neutrophil granulocytes in the lung of horses. We found that: (1). A strong chemotactic activity in the BAL fluid is associated with high levels of dust exposition. (2). In vitro stimulated alveolar macrophages have impaired phagocytosis efficiency and secrete two chemo-attractants specific for neutrophil granulocytes: Interleukin-8 (IL-8) (Wuyts, A., Proost, P., Put, W., Lenaerts, J.-P., Paemen, L., van Damme, J., 1994. Leucocyte recruitment by monocyte chemotactic proteins (MCPs) secreted by human phagocytes, J. Immunol. Meth. 174, 237-247) and macrophage inflammatory protein-2 (MIP-2) (Wolpe, S.D., Sherry, B., Juers, D., Davatelis, G. Yurt, R.W., Cerami, A., Identification and characterisation of macrophage inflammatory protein-2, Proc. Natl. Acad. Sci. USA 86, 612-616; Tekamp-Olson, P., Gallegos, C., Bauer, D., 1990. Cloning and characterisation of cDNAs for murine macrophage inflammatory protein-2 and its human homologues, J. Exp. Med., 172, 911-927; Driscoll, K.E., 1994. Macrophage inflammatory proteins: Biology and role in pulmonary inflammation. Exp. Lung Res., 20, 473-490). This is associated with the appearance of chemotactic activity in the supernatant. These data confirmed our working hypothesis that bronchiolar neutrophilia may be the consequence of a (over)stimulation of pulmonary macrophages leading to expression of cytokines chemotactic for neutrophil granulocytes.

Characterisation of gamma herpesviruses in the horse by PCR.

A polymerase chain reaction (PCR) based on a combination of oligonucleotide primers selected using the octamer frequency disparity method with primers specific for EHV-5 (described by other authors) recognized all of a series of gamma herpesvirus field isolates. This PCR produced only three fragments: (1) one EHV-2-specific; (2) one EHV-5-specific; and (3) a fragment that occurred alone or in combination with the other two. Cloning and sequencing of four different isolates yielding only the last PCR product showed that this corresponds to a deletion/insertion mutant of EHV-2. The fact that this mutant was also plaque-purified from a culture producing all three PCR fragments demonstrated that the virus producing this fragment was distinct from the other two and that this specific DNA fragment was not an artefact due to PCR amplification. These data show that equine gamma herpesviruses are genetically more heterogeneous than previously assumed. The PCR failed to directly detect gamma herpesviruses from the DNA extracted from the same starting material used for the isolation of gamma herpesvirus by cocultivation with indicator cells. This demonstrates that the most reliable method for detection of equine gamma herpesviruses is the cocultivation with indicator cells.

Comparative analysis of equine lymphocyte subsets in whole blood and gradient-purified samples.

In the present study, two methods of lymphocyte preparation, whole blood lysis and Ficoll-Paque separation, prior to FACS analysis were compared. The comparison was done with single and dual-colour staining techniques. Monoclonal antibodies (mAb) against eCD4, eCD5, eCD8 and eMHC class II were used. There was no significant difference in the results obtained by these two methods.

Relation between stallion sperm binding to homologous hemizonae and fertility.

The hemizona assay (HZA) has been developed as a diagnostic test to predict the fertilisation potential of human spermatozoa. The aim of this study was to develop an HZA for stallion spermatozoa and to investigate a possible relationship between fertility and the outcome of the HZA in this species. Equine oocytes were obtained from ovaries collected at a slaughterhouse and by transvaginal, ultrasound-guided follicle aspiration. They were then denuded from cumulus cells and stored in salt solution at 4 degrees C until use. On the day of the experiments the oocytes were bisected, thus providing 2 equal matching hemizonae from each oocyte. Semen samples from Dutch Warmblood stallions with known fertility data were used to assess the number of spermatozoa bound to the outer side of the hemizona after incubation in vitro. Sperm binding to matching hemizonae of a particular stallion was similar and confirmed the feasibility of using the HZA for the horse. Sperm hemizona binding capacity of 10 pairs of stallions was compared by incubating 1 hemizona with the semen of a stallion and the matching hemizona with the semen of another stallion from the same stud farm. Five matching pairs of hemizonae were used for each pair of stallions. There was a significant relationship between the mean number of spermatozoa bound to matching hemizonae and the fertility indices of stallions from each stud farm (P < 0.0001). It is concluded that HZA can be used as a valuable parameter in stallion semen analysis.

[Preliminary results using a combined xylose absorption/hydrogen exhalation test in horses]. / Erste Erfahrungen mit dem Kombinierten Xylose-Absorptions-/Hydrogen-Exhalationstest beim Pferd.

In the present study the breath hydrogen (H2) excretion test was combined with the xylose absorption test in 4 normal horses and 9 clinical patients with chronic diarrhea (n = 3) or chronic weight loss without diarrhea (n = 6). All horses underwent a thorough clinical examination. Laboratory evaluations consisted of haematology and serum biochemistry as well as bacteriological and parasitological examination of feces. In addition, serum electrophoresis and abdominocentesis was performed in all the clinical patients. Gastroscopy was carried out in 6 patients and rectal biopsies were obtained from 4 animals. Two animals were euthanized within 4 weeks after the evaluation. The diagnosis of chronic granulomatous enteritis was confirmed in one of them at necropsy, the cause of weight loss in the other remained unexplained. In the remaining animals, specific causes such as management, parasites, chronic infections and diseases of liver, kidneys and heart were excluded, but no specific diagnosis could be obtained. For the combined xylose absorption/H2 excretion test, the animals were starved overnight and given 0.5/kg/bwt xylose as a 10% solution by stomach tube the next morning. Blood samples were collected for plasma xylose analysis at 30 min intervals for 4 hrs, and breath samples were also collected at 30 min intervals for 8 hrs. With the exception of the animal with granulomatous enteritis, the diseased horses showed only slight alterations in either peak concentrations or times to reach peak levels of plasma xylose. None of the healthy animals showed an increase in breath H2 production after xylose administration, whereas five of the diseased animals showed distinct increases of variable heights. In addition, the diseased horses showed higher fasting breath H2 levels (range 7.5-61.5 ppm) than normal horses (range 0-5 ppm). It is concluded that gastrointestinal disorders might be influenced or even induced by a change in intestinal microbial composition, as evidenced by an increased hydrogenic metabolism.

Videoendoscopic evaluation of the mare's uterus: I. Findings in normal fertile mares.

The new generation of videoendoscopes uses an electronic, instead of an optical, system for image transmission. Advantages over conventional fibre-optic endoscopes include increased image quality, handling robustness and direct display of the image on a TV monitor for multiple simultaneous viewing. In the present study, hysteroscopy was performed on 14 normal fertile Welsh Pony and Thoroughbred mares at various times during the annual and ovarian breeding cycles. Oestrus was characterised by an oedematous, relaxed cervix lying on the floor of the vagina, diffuse oedema of the endometrium and the occasional accumulation of small amounts of clear secretions in the uterine lumen. In dioestrus, the cervix appeared tight and pale and was located in the centre of the vaginal fornix. The endometrium appeared thinner due to the reduction in interstitial fluid, which allowed the underlying blood vessels to become visible, but it still had a glistening surface as a result of endometrial gland secretions. The appearance of the uterotubal papillae reflected the changes in the cervix, being relaxed, pinkish and oedematous during oestrus but remaining pale, tight and erect in dioestrus (between Days 7 and 12 after ovulation). A flexible polythene cannula was passed into the uterine lumen through the working channel of the endoscope for directed collection of uterine fluid for cytological and bacteriological examinations. Site-directed biopsies of the endometrium were taken, with flexible forceps inserted through the working channel of the endoscope, or with rigid forceps passed through the cervix alongside the endoscope. Both methods enabled accurate visual selection of the sampling area but the rigid forceps were preferred due to the larger size of the piece of endometrium recovered.

Videoendoscopic evaluation of the mare's uterus: II. Findings in subfertile mares.

Videoendoscopy of the reproductive tract was performed in 87 Thoroughbred mares with histories of reduced fertility. During hysteroscopy samples for cytological, microbiological and histological examinations were obtained under visual control. Common findings in these broodmares included: (a) endometrial degeneration, as assessed by an uneven distribution or atrophy of endometrial folds and/or a scarred appearance of the endometrium (49 mares, 56%); (b) endometrial cysts of various sizes and locations within the uterus with the most common location being at the base of the uterine horns (48 mares, 55%); (c) fluid accumulation in the uterine lumen (28 mares, 32%). A few mares had transluminal adhesions (7 mares, 8%) and in 2 mares the adhesions appeared to obstruct one uterine horn completely. A solitary discrete lump was detected in the wall of the uterine body in one mare and the suspicion of it being a leiomyoma was confirmed histologically with the aid of a visually directed biopsy sample. Free intraluminal structures were present in the uterine lumen in 3 mares, including one inspissated blood clot and two suspected remnants of resorbing pregnancies. Flexible biopsy forceps and scissors passed through the working channel of the endoscope were used to sever small thin adhesions, but this method proved inadequate for multiple adhesions or cysts. Solitary endometrial cysts were removed by means of conventional rigid biopsy forceps passed alongside the endoscope, although bleeding from the operation site usually limited this type of intervention.

Videoendoscopic evaluation of the mare's uterus: III. Findings in the pregnant mare.

During a 5-year period 65 Pony and 20 Thoroughbred pregnant mares were subjected to videoendoscopic hysteroscopy from 10 to 266 days of gestation. The aims of these examinations were to 1) observe foetal and placental development in vivo (60 Pony and 10 Thoroughbred mares); 2) eliminate one of unicornuate twin conceptuses (9 Thoroughbred mares); 3) recover embryonic foetal and placental tissues non-surgically for experimental purposes (47 Pony mares); 4) induce focal separation of the placenta in late gestation as an experimental model of placentitis (5 pony mares and 1 Thoroughbred mare). It was possible to view the embryo and all constituent extra-embryonic membranes of the conceptus between Days 10 and 87 of gestation without having to perforate the allantochorion. This allowed study of physiological processes such as the coordinated uterine contractions responsible for conceptus motility between Days 7 and 17, active foetal movements, which began as early as Day 34, and invasion of chorionic girdle cells into the endometrium, which occurred between Days 34 and 38. From Day 90, vision of the foetus was reduced or prevented by the increased thickness of the allantochorion. Transendoscopic recovery of the conceptus was successful in all of 10 mares under 30 days of gestation, whereas only 10 of 18 attempts between Days 30 and 45 produced the conceptus without resorting to uterine lavage after initial rupture of the allantochorion. All 9 attempts to eliminate one of unilateral twin conceptuses were unsuccessful and the technique was abandoned as clinically unsuitable. Nevertheless, 9 of 22 (41%) single conceptuses remained viable after one or more hysteroscopic examinations.(ABSTRACT TRUNCATED AT 250 WORDS)

An investigation of the incidence of chronic obstructive pulmonary disease (COPD) in random populations of Swiss horses.

The incidence of Chronic Obstructive Pulmonary Disease (COPD) in Switzerland was determined in three groups of horses selected at random. Group A (97 horses) and Group B (93 horses) had no history of respiratory disease, but Group C (113 horses) had a history of lower respiratory tract disease. All horses were examined by auscultation of the respiratory tract under forced breathing, endoscopic examination of the upper and lower respiratory tract, arterial blood gas analysis at rest and cytological examination of respiratory secretions (RS). Fifty-four per cent of the horses in Group A and 54.8 per cent of Group B were diagnosed as suffering clinical COPD and only 12.4 per cent of horses in Group A and 8.6 per cent in Group B had no detectable abnormalities in their lower respiratory tracts. In Group B, 19.4 per cent were considered to be suffering from parasitic pulmonary disease indicated by elevated numbers of eosinophils in their RS. Pharyngeal lymphoid hyperplasia was exhibited in 70.1 per cent of Group A and 74.2 per cent of Group B. Cytological examination of RS was useful to determine if the secretions found in the trachea originated from the upper or lower respiratory tract. Arterial blood gas analysis revealed no significant differences between healthy horses and those with subclinical or mild COPD.

Prolactin secretion in lactating mares before and after treatment with bromocriptine.

The effect of treating lactating mares with the dopamine agonist bromocriptine was investigated. Seven pony and 4 Thoroughbred lactating mares were given a single intramuscular (i.m.) injection of 100 mg bromocriptine between Days 18 and 28 after foaling when the secretion rate of prolactin was elevated. Prolactin and luteinizing hormone (LH) concentrations were measured in serial peripheral plasma samples taken before and after the treatment and, in 5 of the pony mares, levels of these hormones were also measured in pituitary venous effluent obtained by cannulation of the cavernous sinus. In all the mares, prolactin concentrations fell sharply after treatment (Day 0) and they started to increase again on Day 2 in the Thoroughbreds and Day 3 in the ponies. Pituitary LH secretion rate also declined significantly for 2 days after treatment but there were no measureable influences on cyclic ovarian activity.

[The Weaver syndrome in cattle. Clinical, biochemical and pathologico-anatomic studies in a Braunvieh/Brown Swiss cow with bovine progressive degenerative myeloencephalopathy]. / Das Weaver-Syndrome beim Rind. Klinische, biochemische und pathologisch-anatomische Untersuchungen bei einem Braunvieh-/Brown Swiss-Rind mit boviner progressiver, degenerativer Myeloenzephalopathie (BPDME).

The clinical signs and pathological lesions, consistent with Bovine Progressive Degenerative Myeloencephalopathy (BPDME), are described in a heifer. The animal was an 18-month-old Braunvieh-/Brown Swiss crossbreed with 50% Brown Swiss. Both the sire as well as the dam were related to the bull "B.". The only neurological signs were a posterior ataxia. Mobility and coordination of the front legs were normal. The CK activity of the cerebrospinal fluid was enhanced. There was a moderate to severe degeneration of the white matter consisting of marked axonal degradation and distension and degradation of myelin sheats in all parts of the spinal cord as well as in the medulla oblongata. The lesions were more severe in the descending than in the ascending tracts.