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.     

Pharmacokinetics of sulphadimidine in carp (Cyprinus carpio L.) and rainbow trout (Salmo gairdneri Richardson) acclimated at two different temperature levels

The influence of temperature (10 degrees C and 20 degrees C) on pharmacokinetics and metabolism of sulphadimidine (SDM) in carp and trout was studied. At 20 degrees C a significantly lower level of distribution (Vdarea) and a significantly shorter elimination half-life (T(1/2)beta) was achieved in both species compared to the 10 degrees C level. In carp the body clearance parameter (ClB(SDM)) was significantly higher at 20 degrees C compared to the value at 10 degrees C, whereas for trout this parameter was in the same order of magnitude for both temperatures. N4-acetylsulphadimidine (N4-SDM) was the main metabolite of SDM in both species at the two temperature levels. The relative N4-SDM plasma percentage in carp was significantly higher at 20 degrees C than at 10 degrees C, whereas there was in trout no significant difference. In neither species was the peak plasma concentration of N4-SDM (Cmax(N4-SDM)) significantly different at two temperatures. The corresponding peak time of this metabolite (Tmax(N4-SDM)) was significantly shorter at 20 degrees C compared to 10 degrees C in both carp and trout. In carp at both temperatures, acetylation occurs to a greater extent than hydroxylation. Only the 6-hydroxymethyl-metabolite (SCH2OH) was detected in carp, at a significant different level at the two temperatures. Concentrations of hydroxy metabolites in trout were at the detection level of the HPLC-method (0.02-micrograms/ml). The glucuronide metabolite (SOH-gluc.) was not detected in either species at the two temperatures.     

Effects of thromboxane synthetase inhibition on immune complex glomerulonephritis.

To determine the role of thromboxane A2 in the pathogenesis of experimentally induced immune complex glomerulonephritis, 12 concanavalin A-immunized Beagles were infused with 1 mg of concanavalin A via each renal artery and treated twice daily for 8 days with either 30 mg of CGS 12970/kg, PO, a specific thromboxane synthetase inhibitor, or placebo. The effect of treatment was assessed by measuring endogenous creatinine clearance and urine protein and eicosanoid excretion, and by evaluating changes in glomerular morphometric characteristics. On postinfusion day 8, urine protein, thromboxane B2, and 11-dehydro-thromboxane B2 excretion, glomerular epithelial crescent formation, and glomerular cell proliferation in the CGS 12970-treated dogs were significantly decreased when compared with values in the placebo-treated group. Differences were not observed in endogenous creatinine clearance, urine prostaglandin E2 and 6-keto-prostaglandin F1 alpha excretion, or glomerular polymorphonuclear leukocyte infiltration between groups in this study. These findings suggest thromboxane A2 has a role in the development of immune complex glomerulonephritis and that thromboxane synthetase inhibition may be beneficial in attenuating some of the functional and histological changes associated with immune complex glomerulonephritis.     

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 .     

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 .     

Familial stomatocytosis--hypertrophic gastritis (FSHG), a newly recognised disease in the dog (Drentse patrijshond)

A newly recognised disease, which we have given the provisional name of familial stomatocytosis-hypertrophic gastritis (FSHG), is described in two families of dogs of the Drentse partrijshond breed. The affected dogs consisted of 3 females and 5 males, 3 to 19 (mean 9.5) months of age at admission. The main clinical problems were diarrhoea, icterus, and ataxia and paresis of the pelvic limbs. Laboratory evaluation revealed abnormal red cell shape (stomatocytosis), increased osmotic fragility, haemolytic anaemia, and increased liver enzymes and serum bilirubin. Gastroscopic and histopathologic examination of the gastric mucosa revealed hypertrophic gastritis resembling Ménétrier's disease in man. Histologic findings in the liver were suggestive of progressive liver disease. Cysts were found in the kidneys of the five oldest patients. Electroneurography in 2 dogs revealed polyneuropathy. In the parents of 2 patients (sister and brother), there were no clinical or laboratory abnormalities. An autosomal recessive hereditary defect of lipid metabolism is suspected.     

A Comparison of Injectable Anesthetic Combinations in Horses

Six combinations of injectable anesthetic agents were administered to six adult horses in a Latin square design. The drug combinations were xylazine-ketamine, xylazine-butorphanol-ketamine, xylazine-tiletamine-zolazepam, xylazine-butorphanol-tiletamine-zolazepam, detomidine-ketamine, and detomidine-butorphanol-ketamine. Measured variables were heart rate, respiratory rate, systolic blood pressure, arterial pH (pHa), PaCO2, PaO2, recumbency time, and number of attempts necessary to stand. Quality of induction and recovery, muscle relaxation, and response to stimulus were evaluated subjectively. The horses required significantly more attempts to stand after administration of xylazine-tiletamine-zolazepam, xylazine-butorphanol-tiletamine-zolazepam, and detomidine-ketamine than after xylazine-ketamine, xylazine-butorphanol-ketamine, or detomidine-butorphanol-ketamine. Mean recumbency times varied from 23.0 minutes with xylazine-ketamine to 41.3 minutes with xylazine-butorphanol-tiletamine-zolazepam. There were significant differences in mean heart rates at minute 15, mean respiratory rates at minutes 5, 10 and 15, and mean systolic blood pressures at minute 10 of anesthesia. There were no significant differences in pHa, PaCO2 or PaO2.