Additional diagnostic procedures

This article reviews recent diagnostic procedures that have arisen over the last 10 years. Videoendoscopy of horses on a high-speed treadmill allows observation of some of the changes that take place in a horse's airway during exercise. Measurements of upper airway airflows and transupper airway pressure, the use of an esophageal balloon and a Ventigraph to measure changes in pleural pressure, and pulmonary function testing are new techniques that aid the researcher in understanding the mechanics and pathologic characteristics of airway diseases and help the practitioner in assessing the severity of a problem, measuring response to therapy, and accurately determining a prognosis.     

Participation of H1-receptors in histamine-induced contraction and relaxation of horse coronary artery in vitro.

The mechanisms of histamine-induced contraction and relaxation were investigated in rings isolated from a middle part of the left descending coronary arteries of horses. Intact and endothelium-denuded preparations were compared. Rings of horse coronary arteries contracted in response to histamine in a concentration dependent manner, but some of them relaxed with lower concentrations and contracted with higher concentrations. Removal of the endothelium abolished the relaxation and potentiated the contraction. The pD2 values were 4.70 +/- 0.08 in the rings with intact endothelium and 4.95 +/- 0.08 in endothelium-denuded rings. Histamine-induced contractions in intact and denuded preparations were not affected by an H2-antagonist, cimetidine, but were inhibited by an H1-antagonist, diphenhydramine in non-competitive manner in the rings with endothelium and in competitive manner in denuded rings. After precontraction with PGF2 alpha or norepinephrine, histamine relaxed preparations with intact endothelium (pD2 value, 7.80 +/- 0.11), although histamine-induced relaxations were not observed in denuded preparations. The relaxation was competitively inhibited by diphenhydramine. Relaxing response was significantly attenuated by methylene blue, quinacrine, L-nitro-arginine, gossypol and AA861 but not by indomethacin. These results suggest that the histamine-induced contraction and relaxation in horse coronary arteries are mediated mainly by H1-receptors in the smooth muscle and endothelium, respectively, and H1-receptor activation of endothelial cells may liberate vasodilator substances.     

Genetic Engineering Approaches to Pig Production*

Modern biotechnology promises a number of new applications in animal breeding and production. Although conventional pig breeding has achieved a high level of efficiency and productivity numerous problems have been encountered with animal health and the loss of meat quality. Selection based on phenotypic performance data of individual animals does not take into account the importance of specific genes and their relevance within a complex regulatory system. In most cases it is therefore difficult to trace back the genetic origins of clinically important disorders. The application of genetic engineering techniques in pig production will facilitate diagnosis, improvement of productivity, and animal health by allowing direct genetic manipulation. Attention must be focussed on the physical and genetic analysis of the procine genome. The isolation and characterisation of genes, DNA-markers, polymorphic DNA-fragments, and their chromosomal assignment will be important prerequisites and tools for the elucidation of genetic disorders. Especially the detection of heterozygous carriers of recessive disorders and their elimination from the breeding stock will increase selection accuracy and decrease the generation intervals. But also the rapid and simple detection of infectious diseases, which is sometimes difficult if not impossible at present, will improve animal health and welfare. Although the production of transgenic animals either by DNA-microinjection into zygotes or the use of embryonal stem cells manipulated in vitro is less straightforward than DNA-based diagnosis it will play an important role in the direct manipulation of the porcine genome and genes. Breeding programmes including the use of transgenic livestock have already been developed. There is no doubt that genetic engineering has reached a degree of practical feasibility, allowing it to play an important role in pig breeding in particular and animal production in general.     

Deep Freezing of Boar Semen Packaged in Plastic Bags and Straws

For practical reasons, a large volume (i.e. 5 ml) of frozen boar semen per insemination dose is desirable, but successful freezing has not been achieved, since optimal cooling rates have not yet been established. Post-thaw motility and the acrosome intep'ty of semen from four boars frozen with a programmable freezin machine, in mini-(0.25 ml), maxi-(5 ml) plastic straws and in 10 × 5 cm PVC- or Teflon FEP-plastic bags (0.35 – 0.12 mm thick, 5 ml) was studied. The freezing of the semen was monitored using thermocouples placed in the straws and the bags. The freezing curve started from +5°C, at a rate of −3°C/min, to – 6°C, it was held for 1 min at −6°C, and was followed by further drop to −100°C at a rate of −20°C/min, with subsequent storage in LN₂. The bags had a much shorter freezing point plnteau, compared to the maxi-straws. Post-thaw sperm motility was significantly higher when semen was frozen in mini-straws or in bags than in maxi-straws. The freezing procedure did not cause major acrosomal damages, significantly more normal apical ridges being present in the bags and mini-straws than in the maxi-straws. This in vitro evaluation indicates that the freezing method employed is satisfactory for freezing large volumes of boar semen into plastic bags .     

Distributions of extracellular matrix and carbonic anhydrase-III during bovine palatine ridge development.

The present study describes histological alterations and immunohistochemical distributions of extracellular matrices (ECMs) and the carbonic anhydrase isozyme-III (CA-III) during the period of bovine palatine ridge formation. Morphogenesis of bovine palatine ridges was preceded by epidermal placodes and the mesenchymal condensation (MC). During the early stages of less than 44 cm crown rump length (CRL), fibronectin (FN) was distributed densely in the MC. Strong reactions against type I collagen (C-1) were detected outer to the FN positive site. In the stages of more than 44 cm CRL, FN and C-1 were distributed diffusely in subepithelial mesenchyme. Laminin (LN) and type IV collagen were distributed in the epithelial and endothelial basement membranes (BMs) in all of the stages examined, except in the stage of 7 cm CRL, where LN was not detected only in the BM just beneath the epidermal placode. CA-III was detected in basal epithelial cells except for palatine ridge rudiments in the stages of more than 21 cm CRL. It is suggested that the expressions of LN and CA-III might play a role in the spatial determination of rudiments of bovine fetal palatine ridges.     

Changes in the MHC Class II+ Dendritic Cell Population of Ovine Skin In Response to Orf Virus Infection

Abstract— Class II+ dendritic cells were widely distributed throughout normal ovine skin in two main locations: a) in or immediately adjacent to the epidermis and epidermal appendages and b) in the vicinity of the blood vessels. They are unlikely to represent a homogeneous population particularly since Langerhans cells, which previously have been found throughout the epidermal appendages, were located only in the epidermis using acetylcholinesterase staining. Following infection with orf virus, a dense mass of closely associated class II+ dendritic cells develops in the exposed necrotising dermis, adjacent to infected hair follicles and under infected degenerating epidermis. These cells interact and appear to form a barrier to invasion, a framework for immune defence and a template for subsequent epidermal repair; they seem to provide the basis of a highly integrated local dermal defence system. Résumé— Des cellules dendritiques de classe II+étaient largement réparties dans la peau ovine normale dans deux principales zones a) dans l'épiderme et les annexes épidermiques ou dans leur voisnage immédiat b) au voisinage des vaisseaux sanguins. Il est peu probable qu'elles représentant une population homogène particulièrement parce que les cellules de Langerhaps, qui ont été découvertes précédemment dans l'ensemble des annexes épidermiques, furent localisées seulement dans l'épiderme en utilisant une coloration à l'acétylcholinesterase. Après une infection par le virus de l'ecthyma, il se forme une masse dense de cellules dendritiques de classe II+étroitement associées, dans le derme nécrotique atteint, adjacente aux follicules pileux infectés et sous l'épiderme dégénératif infecté. Ces cellules interagissent et apparaissent former une barrière à l'invasion, un cadre pour les défenses immunitaires et un patron pour la réparation épidermique ultérieure; elles semblent fournir les bases d'un système de défense dermique local hautement intégré. Zusammenfassung— Klasse II+-Dendritenzellen waren in der gesamten Haut von normalen Schafen in vorwiegend zwei Bereichen verbreitet: a) in oder unmittelbar neben der Epidermis und der epidermalen Anhangsgebiete und b) in dor Nähe dar Blutgefäße. Sie stellen wahrscheinlich keine homogène Population dar, da die Langerhanszellen, die früher übarall in den epidermalen Anhangsgebilden nachgewiesen wurden, bei Acetylcholinesterase-Färbung nur in der Epidermis zu finden waren. Nach einer Orf-Virus-Infektion formiert sich eine dichte Masse aus eng verbundenen Klasse II+-Dendritenzellen in der betroffenen nekrotisierenden Dermis unmittelbar neben den infizierten Haarfollikeln und unter der infizierten, degenerierenden Epidermis. Diese Zellen stehen untereinander in Verbindung und bilden anscheinend eine Barrière gegen die Invasion, ein Gorüst für die Immunabwehr und einen Ausgangs punkt für die anschließenden Reparaturvorgänge in der Epidermis. Sie scheinen als Basis eines hochentwickelten lokalen Abwehrsystems der Haut zu fungieren. Resumen Células dendríticas de class II+ se observaron en gran cantidad en la piel de la oveja especialmente en dos localizaciones: a) en la epidermis, en la dermis muy próxima a la epidermis y en anejos cutáneos y b) en las proximidades de los vasos sanguíneos. No parece tratarse de una población homogénea de células puesto que las células de Langerhans, que previamente se habían encontrado en los anejos epidérmicos, se encontraron únicamente en la epidermis utilizando técnicas de detección del acetilcol-inesterasa. Después de la infección con el virus del ectima contagioso ovino se observó una masa de células dendríticas de clase II, dispuestas de forma muy densa, en las proximidades de la dermis necrosada y de los folículos pilosos y de la epidermis infectada. Eatas células interaccionan entre si y parecen formar una barrera contra la invasión, una red inmunitaria de defensa y participar en la reparación de la epidermis; parece ser que esta población de células dendriticas son la base de un sistema de defensa dérmico local altamente integrado.     

Artificial Insemination with Frozen Semen in Ewes at Different Times of the Breeding Season

Totally 13575 ewes of two different breeds, Dala and Spel, were inseminated with semen, frozen in straws and thawed at 70°C for 8 sec. An insemination dose of 0.2 ml containing approx. 150 × 10⁶ spermatozoa with at least 45 to 50% progressive motility was imerted 5 to 12 mm into the cervix. The insemination was performed once between 12 and 30 h after the onset of heat. The NR rate of the Dala ewes increased significantly during the season. The NR rate of the ewes inseminated before 15. November was 44.3%, from 15. to 20. November 52.2%. from 20. to 25. November 55.3% and from 25. November and later 61.4%. The corresponding values for the ewes of the Spel breed were 57.3, 58.7, 61.5 and 71.0% respectively, and only the difference between the two last values was statistically significant. The difference between the fertility of the two breeds was significant within each of the periods .     

Vitamin deficiencies in cattle

Deficiencies of vitamins A, D, K, E and thiamin can cause severe limitations in beef production. In particular, vitamin A and E can be common causes of lost profit, secondary to limitations of reproductive and growth potential. Prolonged dry periods will reduce available A and E in pasture forage, as can ensiling and prolonged storage of harvested feedstuffs. Polioencephalomalacia is a thiamin responsive disorder, associated with high concentrate feeding and lush pastures. Antimetabolites, such as amprolium, will cause thiamine deficiency when fed in excess. Recent information has shown improved performance with supplemental beta carotene and niacin. The positive responses in reproductive performance, noted with cattle fed supplemental beta carotene, was independent of vitamin A. Supplementation of vitamins above National Research Council recommendations can be justified. However, proper evaluation of feed and animal status, and documentation of a response to supplementation is necessary before diagnosing deficiencies of specific nutrients.     

A novel HPLC method for glutathione-insulin transhydrogenase assay using fluorescence labeled insulin

The activity of rat liver glutathione-insulin transhydrogenase (GIT) was measured by HPLC. The degradation of fluorescein isothiocyanate-I (FITC-I)-labeled insulin is separated into several peaks, which are bound different amount of FITC-I. We selected mono-fluorescein-thiocarbamylated insulin to estimate the decrease of insulin content and it became possible to assay GIT activity. This novel method was time-saving and simple, and this system could utilize instead of previous method.