Detection of antibodies against Pasteurella multocida using immunohistochemical staining in an outbreak of rabbit pasteurellosis

To detect serum antibody against Pasteurella multocida (P. multocida) in infected rabbits. a modified immunoperoxidase assay was applied. An outbreak of P. multocida infection in rabbits started from sudden death. The infected rabbits had severe fibrinous and purulent pneumonia with hemorrhage, and a large number of P. multocida (A:12) was isolated from the trachea and lungs of the animals. Antibodies of IgM and IgG to P. multocida were assessed by immunohistochemical staining using the sera of the animals as primary antibodies and applying them to formalin-fixed, paraffin-embedded sections of P. multocida attached to calf fibrin. IgM antibodies to P. multocida were first detected 7 days after the onset of the disease. IgG antibodies began to rise on the 7th or 14th day. These results suggested that the modified immunoperoxidase assay could detect antibodies against P. multocida.     

Pharmacokinetic/pharmacodynamic modelling of robenacoxib in a feline tissue cage model of inflammation

Pelligand, L., King, J. N., Toutain, P. L., Elliott, J., Lees, P. Pharmacokinetic/pharmacodynamic modelling of robenacoxib in a feline tissue cage model of inflammation. J. vet. Pharmacol. Therap.  35 , 19–32. Robenacoxib is a novel nonsteroidal anti‐inflammatory drug developed for use in cats. It is a highly selective COX‐2 inhibitor. Results from previous feline studies showed that, despite a short half‐life in blood, the effect of robenacoxib persisted for 24 h in clinical studies. A tissue cage model of acute inflammation was used to determine robenacoxib’s pharmacokinetics and its ex vivo and in vivo selectivity for COX‐1 and COX‐2 using serum TxB₂ and exudate PGE₂ as surrogate markers for enzyme activity, respectively. After intravenous, subcutaneous and oral administration (2 mg/kg), the clearance of robenacoxib from blood was rapid (0.54–0.71 L·h/kg). The mean residence time (MRT) in blood was short (0.4, 1.9 and 3.3 h after intravenous, subcutaneous and oral administration, respectively), but in exudate MRT was approximately 24 h regardless of the route of administration. Robenacoxib inhibition of COX‐1 in blood was transient, occurring only at high concentrations, but inhibition of COX‐2 in exudate persisted to 24 h. The potency ratio (IC₅₀ COX‐1: IC₅₀ COX‐2) was 171:1, and slopes of the concentration–effect relationship were 1.36 and 1.12 for COX‐1 and COX‐2, respectively. These data highlight the enzymatic selectivity and inflamed tissue selectivity of robenacoxib and support the current recommendation of once‐daily administration.     

Infectious bovine keratoconjunctivitis II. Antibodies in lacrimal secretions of cattle naturally or experimentally infected with Moraxella bovis.

One or both eyes of 20 calves were inoculated one or more time with variou(s combinations of microorganism (live oor killed Moraxella bovis, infectious bovine rhinotracheitis virus, bovine adenovirus, bovine parainfluenza-3 virus and Mycoplasma bovoculi) by conjunctival instillation or direct inoculation of the conjunctivea or cornea. The eyes of all the calves received natural or artificial ultraviolet irradiation. Neither the adenovirus nor parainfluenza-3 virus became established in the eye or produced keratoconjunctivitis. Both M. bovis and infectious bovine rhinotracheitis virus became established in the bovine eye and produced disease. Subconjunctival or intracorneal inoculation of M. bovis caused a severe disease, simulating natural infectious bovine keratoconjunctivitis. Only the intracorneal inoculation of mycoplasma produced severe keratoconjunctivits. Eyes that on initial exposure to M. bovis became severly inflamed were more resistant to a second or third exposure to M. bovis, presumably by enhanced local defence mechanisms.     

Decreased serum apolipoprotein A-I concentrations in cows infected with Salmonella typhimurium.

Serum apolipoprotein A-I concentrations in cows infected with Salmonella Typhimurium were evaluated to assess its relevance in salmonellosis. Apolipoprotein A-I has been shown in rats to be secreted by the intestine as well as the liver. Clinical symptoms such as diarrhea revealed an outbreak of salmonellosis in 22 cows on a farm, and sera were obtained at 6 (acute phase), 16, 28 (convalescent period) and 42 d (postconvalescent period) after the outbreak. Apolipoprotein A-I concentrations (mean +/- SD, mg/mL), determined by ELISA, were 0.598 +/- 0.497 (day 6), 0.111 +/- 0.060 (day 16), 0.432 +/- 0.311 (day 28) and 0.727 +/- 0.516 (day 42). Compared with the concentration at day 42, those at 16 and 28 d were significantly (P < 0.01, P < 0.05) lower, but that at day 6 was not. The serum concentration of apolipoprotein B-100 (of liver origin in cattle) was unaltered during the course of salmonellosis. The concentration of apolipoprotein A-I was positively correlated with those of serum total cholesterol (r = 0.589, P < 0.01) and phospholipids (r = 0.590, P < 0.01). These results suggest that apolipoprotein A-I in cattle is in part of intestinal origin, and also that its decreased serum concentration in salmonellosis can be attributed to the reduced intestinal synthesis or secretion of this apolipoprotein. Moreover, as a potential carrier for dietary lipids such as cholesterol, determination of serum apolipoprotein A-I concentration is suggested to be useful when assessing the nutritional status of the affected cows.     

Isolation of Mycobacterium species from raw milk of pastoral cattle of the Southern Highlands of Tanzania

A study to determine the secretion of Mycobacterium spp. in milk from indigenous cattle was carried out in pastoral cattle reared in the Southern Highlands of Tanzania. The study was aimed at elucidating the dangers associated with milk-borne zoonoses in a society where milk is normally consumed raw. Out of 805 milk samples, 31 (3.9%) were positive for mycobacteria. There was a preponderance of atypical mycobacteria (87%) whereas only two isolates (6.5%) were confirmed as M. bovis. Atypical mycobacteria included: M. terrae (n=7), M. fortuitum (n=2), M. flavescens (n=13), M. gordonae (n=1) and M. smegmatis (n=4). Although the number of M. bovis positive samples was low, the habit of pooling milk may still pose great public health dangers to milk consumers in this part of the world. Moreover, isolation of atypical mycobacteria should also be considered to be a danger to human health in countries such as Tanzania, where the number of people with lowered immunity due to HIV infection is on the increase.     

Effect of dietary protein content on growth performance, feed utilization and carcass composition in the Australian freshwater crayfish, Cherax albidus Clark and Cherax destructor Clark (Decapoda, Parastacidae)

Cherax albidus (A) and Cherax destructor (D) male juveniles (mean weight 0.95 ± 0.03 g) were reared for 20 weeks on isoenergetic diets containing 150 g kg^?1 protein (A 15, D15) or 300 g kg^?1 protein (A30, D30). Mean weight, percentage weight gain, and specific growth rate (%) were substantially higher for both species on the 300 g kg^?1 protein diet. Mean percentage weight gain ranged from 2.39% day^?1 (D15) to 17.59% day^?1 (A30). A maximum weight of 33.81 g was attained by C. albidus on the higher protein diet. The most effective utilization of food was observed in C. albidus when fed the higher protein diet (food conversion ratio, 0.79; protein efficiency ratio, 4.21; apparent net protein utilization, 44.64%). Carcass composition was influenced by feed type. The higher protein diet resulted in an increase in carcass protein and ash and a decrease in carcass lipid and energy relative to the low-protein diet (150 g kg^?1 protein diet –C. albidus: 37.15% protein, 15.00% lipid, 25.20% ash, 15.55kJ g^?1 energy; C. destructor: 38.10% protein, 15.43% lipid, 25.70% ash, 15.65kJ g^?1 energy; 300 g kg^?1 protein diet –C. albidus: 46.10% protein, 8.71% lipid, 27.36% ash, 14.94kJ g^?1 energy; C. destructor: 42.99% protein, 8.56% lipid, 26.44% ash, 14.71kJ g^?1 energy). Carcass moisture and calcium were not affected by feed type. The time spent in the intermoult phase of growth was highly dependent on the premoult weight and varied according to diet and to species. A comparison of animals of similar weight (< 8 g) revealed that elevated dietary protein caused a reduction in the intermoult period by 11 days in C. albidus and 7 days in C. destructor. The moult increment, however, was independent of animal weight, and the highest percentage weight increment occurred for C. albidus fed the 300 g kg^?1 protein diet (per cent weight increase; A15, 33.1%; A30, 61.3%; D15, 31.2%; D30, 56.5%). Dietary induced morphological changes were also recorded. Animals of a standard carapace length had significantly larger abdomens (both species) and larger claws (C. albidus) when fed the higher protein diet.     

Telomere length and telomerase activity in canine mammary gland tumors

OBJECTIVE: To measure telomere length and telomerase activity in naturally occurring canine mammary gland tumors. SAMPLE POPULATION: 27 mammary gland tumor specimens obtained during resection or necropsy and 12 mammary gland tissue specimens obtained from healthy (control) dogs. PROCEDURE: Telomere length in tissue specimens was measured by use of restriction endonuclease digestion and Southern blot analysis. Telomerase activity was measured by use of a telomeric repeat amplification protocol assay. RESULTS: Telomere length in mammary gland tumors ranged from 11.0 to 21.6 kilobase pairs (kbp; mean +/- SEM, 14.5+/-0.5 kbp) but did not differ among tumor types. Telomeres in mammary gland tumors were slightly shorter than in normal tissue specimens, but telomere length could not be directly compared between groups, because mean age of dogs was significantly different between groups. Age was negatively correlated with telomere length in control dogs but was not significantly correlated with length in affected dogs. Telomerase activity was detected in 26 of 27 mammary gland tumors and in 4 of 12 normal tissue specimens. However, telomerase activity and telomere length were not correlated in tumor specimens. CONCLUSIONS AND CLINICAL RELEVANCE: Telomere length is maintained in canine mammary gland tumors regardless of the age of the affected dog. Measurement of telomere length may be a useful tool for monitoring the in vivo effects of telomerase inhibitors in dogs with tumors.     

Proteomics: A new tool in bovine claw disease research

Claw horn disruption (CHD) is a common underlying cause of lameness in dairy cattle which leads to compromised animal welfare and production losses. Despite an intense research effort over the last two decades, progress in reducing the prevalence of lameness due to CHD has been limited. In addition to current research strategies there is a need to develop novel approaches and methods that expand understanding of the disease mechanisms involved in CHD. The objectives of the present study were to explore the potential of liquid chromatography tandem mass spectrometry (LC-MS/MS) in mapping protein expression in three different bovine claw tissues, and to provide a relevant functional annotation of the proteins characterized in these tissues. LC-MS/MS was used to characterize protein expression in coronary band skin (C), claw dermal (D) and lamellar (L) tissues from two heifers. A total of 388 different proteins were identified, with 146 proteins available for identification in C, 279 proteins in D and 269 proteins in L. A functional annotation of the identified proteins was obtained using the on-line Blast2GO tool. Three hundred and sixteen of the identified proteins could be subsequently grouped manually to one or more of five major functional groups related to metabolism, cell structure, immunity, apoptosis and angiogenesis. These were chosen to represent basic cell functions and biological processes potentially involved in the pathogenesis of CHD. The LC-MS/MS-based proteomic analysis presented here is the largest published survey, so far, of the bovine claw tissue proteome.     

Swine fever. Immunisation of piglets

Vaccination against Swine Fever using the CL Chinese strain can be done in 7-day-old piglets if they are born of non-immune sows. The simultaneous weaning and vaccination emphasises the safety of this strain. The excellent immunity observed confirms the immunocompetence of 7-day-old piglets. In piglets born of immune sows and also weaned at 7 days, passive protection can extend beyond the age of 2 months if the sow is vaccinated several months prior to gestation. The immune level of the piglets would seem to depend on the interval between vaccination of the sow and farrowing and can be attributed to the quality of the antibodies transmitted by the colostrum. Piglets born of sows vaccinated 10 months prior to farrowing can be vaccinated as early as 5 weeks; the protection percentage observed at the age of about 6 months is over 80%. A booster injection at this age then confers immunity to future breeders throughout their economic life, i.e. 4 years in the reported experiment.     

Relationships between disease levels on seed tubers,on crops during growth and in stored potatoes. 1. Introduction and black scurf

Summary In the Introduction general methods are described to assess inocolum of several potato diseases on seed tubers; on the progeny tubers during growth, at harvest and after 4 different storage regimes. Assessments were made in up to 26 commercial King Edward crops in each of 5 years, and up to 13 crops from ‘healthier’ seed, derived from stem cuttings, in 4 years. Results are reported from the work on black scurf (Rhizoctonia solani). Stem canker incidence was more closely related to the incidence of sclerotia on seed tubers than was the frequency ofRhizoctonia hyphae on growing tuber surfaces. Incidence of sclerotia on stored tubers was better related to the amount of surface colonization during growth than that of stem canker. Assessments on harvested and stored progeny of ‘healthier’ seed were correlated with those on adjacent commercial crops, indicating spread or, more likely, a common source of inocolum.

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Zusammenfassung In einer Studie wurde das Auftreten verschiedener Krankheiten im Erntegut der Sorte King Edward untersucht, die zwischen 1971 und 1975 in dem Fenland-Gebiet von Ostengland gewachsen war. Jedes Jahr wurden zwischen 15 und 26 Proben von Marktware untersucht und zwischen 1972 und 1975 wurde ‘gesünderes’ Pflanzgut (das von Stecklingen abstammte) auf dem selben Feld wie einige der Proben der Marktware gepflanzt (Tab. 1). Die Feststellung des Inokulums oder der Krankheit wurde an Pflanzgutproben, an Pflanzen w?hrend des Wachstums und an Knollen der Nachkommenschaft zum Erntezeitpunkt genommen. Geerntete Knollen wurden auch einheitlich verletzt und bei 3°C und 10°C gelagert, mit oder ohne zweiw?chige Wundheilperiode bei 15°C. Nach 3–4 Monaten erfolgte die Feststellung des Krankheitsbefalls. Die Auswertung der Ergebnisse wurde durch Regression zwischen allen m?glichen Paaren der Ergebnisse für jede Krankheit durchgeführt, wobei jedes Jahr sowohl getrennt als auch alle Jahre kombiniert genommen wurden. Die Regressionen wurden auch für das ‘gesündere’ Erntegut, das mit der entsprechenden Marktware übereinstimmte, durchgeführt. Rhizoctonia solani wurde auf dem Pflanzgut und auf dem nachfolgenden Erntegut als Prozentbefall mit Sklerotien angegeben, der Myzelbelag auf der Knollenoberfl?che rund um die Augen wurde durch mikroskopische Untersuchung von Augenstücken festgestellt und das Auftreten von Weisshosigkeit wurde w?hrend des Wachstums gesch?tzt. Das durchschnittliche Auftreten von Inokulum oder Krankheitsbefall auf Marktware (Tab. 2) zeigte, dass 1975 der Befall der Augen am h?ufigsten war und dass 1971 und 1975 der Krankheitsbefall im Lager am gr?ssten war. ‘Gesünderes’ Pflanzgut hatte gew?hnlich weniger Pocken als der Durchschnitt der Marktware aber der Befall der Augen der Tochterknollen w?hrend des Wachstums und zur Ernte war gr?sser und auf gelagerten Knollen wurden mehr Pocken gefunden (Tab. 3). Unter den nassen Erntebedingungen 1974 traten mehr Pocken auf den bei 10°C gelagerten Knollen auf als auf den bei 3°C, aber es ergaben sich keine Unterschiede in anderen Jahren. Die Signifikanz der Regressionen zwischen den Erhebungen, die nach verschiedenen Methoden oder zu verschiedenen Zeitpunkten (Tab. 4) gemacht wurden und Diagramme der signifikanten Regressionen in einzelnen Jahren (Abb. 1) zeigten, dass makroskopische und mikroskopische Bestimmungen auf dem Pflanzgut (Abb. la) und Befall der Schale rund um die Augen und Weisshosigkeit w?hrend des Wachstums (Abb. 1d) in allen Jahren miteinander verbunden waren. Die Weisshosigkeit stand h?ufiger in Beziehung zum Ausmass des Pockenbesatzes des Pflanzgutes (Abb. 1b) als der Befall der Augen (Abb. 1c). Dies führt zu der Annahme, dass die meisten Stengelinfektionen vom Inokulum des Pflanzgutes ausgingen w?hrend der Befall der Schale rund um die Augen auch durch andere Inokulumquellen (z.B. den Boden) beeinflusst wurden. Der Befall der Augen zur Ernte stand in jedem Jahr in Beziehung zum Befall w?hrend des Wachstums (Abb. 1e) und in 4 Jahren stand das Ausmass des Pockenbesatzes auf gelagerten Knollen in Beziehung zum Augenbefall w?hrend des Wachstums und zur Ernte (Abb. 1f und 1g). Die mikroskopische Untersuchung w?hrend des Wachstums gibt daher eine vernünftige Sch?tzung des Krankheitsbefalls im Lager. Wenn die Erhebungen des ‘gesünderen’ Pflanzgutes mit denen gleichartiger Marktware verglichen wurden (Abb. 2), ergaben sich signifikante Verbindungen für den Augenbefall w?hrend des Wachstums und zur Ernte, für den Pockenbesatz auf gelagerten Knollen, aber nicht für die Weisshosigkeit. Dies weist auf eine Verbreitung des Inokulums, auf die Bedeutung der allgemeinen Bodenbeschaffenheit oder, wahrscheinlicher, eine gemeinsame Quelle des Inokulums, vielleicht im Boden hin.

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Résumé Une étude a été entreprise sur la variété King Edward, pour conna?tre l'incidence de différentes maladies. Cette étude s'est déroulée de 1971 à 1975 dans la région de Fenland en Angleterre Orientale. Chaque année, on a examiné entre 15 et 26 lots commerciaux; entre 1972 et 1975 des semences ‘plus saines’ (provenant de bouturage) ont été plantées dans le même champ que les lots commerciaux (tableau 1). Les estimations d'inoculum ou de maladie ont été faites sur des échantillons de tubercules de semence, sur des plantes en course de croissance, et sur les tubercules-fils à la récolte. Les tubercules-fils récoltés ont été également endommagés de manière uniforme et conservés à 3°C et 10°C, avec ou sans période initiale de cicatrisation des blessures de 2 semaines à 15°C. L'incidence de la maladie a été estimé après 3–4 mois. Les résultats ont été analysés par régressions entre toutes les paires possibles de données pour chaque maladie, en prenant séparément chaque année et toutes les années combinées. Les régressions ont aussi été faites à partir des estimations provenant des lots ‘plus sains’, et des estimations correspondantes provenant des lots commerciaux adjacents. L'estimation du rhizoctone a été faite par notation du pourcentage de sclérotes sur les tubercules de semence et sur les tubercules-fils en conservation. La colonisation de la peau, autour des yeux, par du mycelium deRhizoctonia solani a été déterminée par examen microscopique au niveau des yeux. L'incidence des nécroses sur tige a été estimée au cours de la croissance. L'incidence moyenne de l'inoculum ou de la maladie sur les lots commerciaux (tableau 2) a montré que la colonisation du tubercule au niveau des yeux a été plus fréquente en 1975, et que l'incidence de la maladie en cours de conservation a été plus forte en 1971 et 1975. Les tubercules de semence ‘plus saine’ ont eu moins de rhizoctone que la moyenne des lots commerciaux, mais la colonisation des tubercules-fils, au niveau des yeux pendant la période de croissance et à la récolte a été plus grande. On a également trouvé plus de sclérotes sur les tubercules conservés (tableau 3). En 1974, les conditions de récolte étant plus humides, il y avait plus de sclérotes sur les tubercules conservés à 10°C que sur ceux conservés à 3°C; mais, pour les autres années, il n'y avait aucune différence entre les traitements. La signification des régressions entre les estimations faites par différentes méthodes ou à différentes époques (tableau 4), et les graphiques relatifs aux régressions significatives pour chacune des années (Fig. 1) ont montré qu'il y avait toutes les années des relations entre les estimations macroscopiques et microscopiques sur la semence (Fig. 1 a), la colonisation de la peau autour des yeux et les nécroses des tiges en cours de végétation (Fig. 1d). Les nécroses sur tige étaient plus fréquemment en rapport avec les sclérotes portés par la semence (Fig. 1b) qu'ac la colonisation au niveau des yeux du tubercule (Fig. 1c). Les auteurs suggèrent donc que la plupart de la contamination de la tige provient de l'inoculum présent sur le tubercule de semence mais que la colonisation de la peau autour des yeux est également influencée par d'autres sources d'inoculum (par ex. le sol). Chaque année, la colonisation des yeux à la récolte était en relation avec celle en cours de croissance (Fig. 1c). Pour les 4 années, la quantité de sclérotes sur les tubercules conservés était en rapport avec l'incidence de la colonisation au niveau des yeux pendant la croissance et à la récolte (Figures 1f et 1g). Les estimations faites en cours de croissance donne donc une estimation correcte de l'incidence de la maladie en cours de conservation. Quand on compare les résultats des lots ‘plus sains’ et ceux des lots commerciaux adjacents, il y a des associations significatives pour la colonisation des yeux durant la croissance et à la récolte, pour les sclérotes sur les tubercules conservés, mais pas pour les nécroses sur tige. Ceci suggère la dispersion de l'inoculum, l'importance des conditions de sol ou plus vraisemblablement la présence d'une source commune d'inoculum au niveau du sol.