Molecular epidemiology of Pasteurella multocida in turkeys.

Pasteurella multocida isolated from turkeys during an outbreak of fowl cholera was characterized by serotype and heterogeneity of genes encoding rRNA (ribotype) to investigate the epidemiology of the organism. Isolates were collected between October 1985 and July 1986. The M9 or Clemson University fowl cholera vaccine-like strain was detected in 17% of the flocks with fowl cholera. One particular strain, isolated only from breeder flocks, was recovered from 7 of the 10 breeder flocks examined in this study. Intracompany transmission appeared to be common, implying a failure in biosecurity. Circumstantial evidence indicated that in the field; the incubation period of P multocida in a turkey flock may be between 2 to 7 weeks. Wildlife did not appear to be an important reservoir of P multocida for turkeys during this study period. Ribotyping results tended to discount several of the possible interflock transmissions, as suggested by examination of serotyping results alone; however, serotyping in combination with ribotyping proved helpful in understanding the epidemiology of P multocida in turkeys.     

Sedative and cardiovascular effects of romifidine, alone and in combination with butorphanol, in the horse

The behavioural and sedative effects of intravenous (iv) romifidine (40 and 80 μg/kg bodyweight [bwt]) alone or in combination with iv butorphanol (50 μg/kg bwt) were investigated in four ponies and one Thoroughbred horse. Apparent sedation, as judged by the lowering of the head, and by the response to imposed touch, visual and sound stimuli was assessed. The combination with butorphanol reduced the animals' response to imposed stimuli when compared with the effect of the same dose of romifidine alone. Following the administration of romifidine/butorphanol combinations muzzle tremor was noted and some animals attempted to walk forward. In a separate series, the cardiopulmonary effects of iv romifidine (80 μg/kg bwt) alone, or in combination with butorphanol (50 μg/kg bwt) were investigated. Romifidine and the romifidine/butorphanol combination caused similar cardiovascular changes, these being bradycardia with heart block, and hypertension followed by hypotension. Romifidine caused a transient decrease in arterial oxygen tensions and arterial carbon dioxide tensions had increased significantly by the end of the 90 min recording period. Romifidine/butorphanol combinations produced significantly higher arterial carbon dioxide tensions during the first 15 mins after drug administration than did romifidine alone. Butorphanol at 50 μg/kg bwt iv reduced the response to imposed stimuli in horses sedated with romifidine. The combination produced no cardiovascular changes beyond those induced by romifidine alone, but did increase the degree of respiratory depression.     

Confirmation of rinderpest in experimentally and naturally infected cattle using microtitre techniques

Summary Virulent rinderpest virus was detected by immunoperoxidase staining of microtitre bovine kidney cell cultures within 24 to 48 hours of inoculation with prescapular lymph node and spleen homogenates from experimentally infected steers. Rinderpest virus specific cytopathic effects were evident from 48 hours in microtitre plates and from 72 hours in rolled tube cultures. Nasal and ocular secretions collected from cattle naturally infected with rinderpest and inoculated into bovine kidney cell cultures did not readily yield cytopathic virus in both tubes and microtitre plates, but immunoperoxidase staining of microtitre cultures on the fourth day of inoculation detected replication of virus in cultures inoculated with ocular and nasal secretions from seven of 17 cattle tested.

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Resumen Se detectó el virus virulente de mediante la tinción con inmunoperoxidasa de cultivos de células de ri?ón bovino en bandejas de microtitulación, después de la inoculación de estos con suspensiones homogenizadas de ganglios linfáticos preescapulares y de bazo provenientes de novillos infectados experimentalmente. El efecto citopático del virus de la peste bovina fue evidente desde las 48 horas en bandejas de microtitulación y desde las 72 horas en tubos de cultivo giratorios. Secreciones oculares y nasales colectadas de ganado infectado en forma natural con la peste bovina e inoculadas en cultivos de células de ri?ón bovino, no mostraron efecto citopático fácilmente en tubos giratorios o bandejas de microtitulación, pero la tinción de las bandejas con inmunoperoxidasa reveló replicación del virus a partir del cuarto día de inoculación con secreciones oculares y nasales en siete de los 17 animales examinados.

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Résumé Un virus bovipestique virulent a été décelé par le test de coloration à l'immunoperoxydase de cellules rénales bovines en culture dans des plaques de microtitrage et infectées 48 heures plus t?t avec des homogénats de ganglions lymphatiques et de rate provenant de bouvillons infectés expérimentalement. Les effets cytopathogènes du virus étaient évidents au bout de 48 h dans les plaques de microtitrage et 72 h dans les tubes en rollers. Les sécrétions nasales et oculaires prélevées sur du bétail infecté naturellement par la peste bovine et inoculées sur des cellules rénales bovines n'ont pas toujours montré d'effet cythopathogène aussi bien dans les tubes que dans les plaques de microtitrage. Cependant, la coloration à la peroxydase au jour 4 après l'inoculation a permis de déceler la présence de virus dans 7 cas sur 17.

     

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.     

Effect of short-term calf removal at three stages of a follicular wave on fate of a dominant follicle in postpartum beef cows.

The hypothesis that ovulation in response to short-term (48 h) calf removal (CR) is dependent on the developmental stage of the dominant follicle was tested in two studies. The objective of Exp. 1 was to characterize the fate of a dominant follicle following 48-h CR on d 2, 4, or 8 of a postpartum follicular wave. Ovaries of 61 beef cows were examined daily by transrectal ultrasonography starting at d 20 to 21 postpartum. Treatments were no CR (n = 14) and CR on d 2 (n = 12), 4 (n = 16), or 8 (n = 10) of first detected follicular wave. Percentage of cows that ovulated a dominant follicle following treatment was not different among groups (P = 0.62). Maximum size of dominant follicles was larger in cows that ovulated (P = 0.002) than in cows that did not ovulate. The objectives of Exp. 2 were 1) to determine whether a follicular wave could be synchronized in anestrous cows following injection of 1 mg of estradiol benzoate (EB) and 200 mg of progesterone (P4; EB + P4); 2) to characterize the fate of dominant follicles following 48-h CR at three stages of a synchronized follicular wave; and 3) to determine whether estrous cycles of normal length followed ovulation in cows pretreated with EB + P4. Ovaries of 50 anestrous beef cows were examined daily as in Exp. 1. Treatments were sesame oil (SO) injected (i.m.) on d 25 postpartum and no CR (n = 9); EB + P4 and no CR (n = 9); EB + P4 and CR on 6 (n = 12), 8 (n = 9), or 12 (n = 11) d after injection. The EB and P4 injections were given on d 25 postpartum. Variability in day of emergence of subsequent follicular waves was lower in cows receiving EB + P4 than in SO-injected cows (P < 0.05). The percentage of cows that ovulated was not different (P = 0.16), but CR increased the percentage of cows that ovulated when groups that received EB + P4 were compared to the EB + P4 group that did not have CR (53.1 vs 11.1%, respectively; P < 0.05). Maximum diameter of dominant follicles was larger (P = 0.05) in ovulatory follicles. The luteal phase was longer (P < 0.03) in cows receiving EB + P4 injection (10.6 +/- 1.2 d) than in cows receiving SO (4.4 +/- 2.2 d). In summary, the maximum size of ovulatory follicles was greater than that of nonovulatory follicles, the ovulatory response of postpartum anestrous cows was maintained through d 8 of a follicular wave, synchronization of follicular waves was accomplished in postpartum cows using EB + P4, and the subsequent luteal phase length was increased in animals that were administered EB + P4.     

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.