Inter-laboratory and inter-assay comparison on two real-time PCR techniques for quantification of PCV2 nucleic acid extracted from field samples

Several real-time PCR assays for quantification of PCV2 DNA (qPCR) have been described in the literature, and different in-house assays are being used by laboratories around the world. A general threshold of 10(7) copies of PCV2 per millilitre serum for postweaning multisystemic wasting syndrome (PMWS) diagnosis has been suggested. However, neither inter-laboratory nor inter-assay comparisons have been published so far. In the present study, two different qPCR probe assays used routinely in two laboratories were compared on DNA extracted from serum, nasal and rectal swabs. Results showed a significant linear association between the assays (p<0.0001), and a systematic difference of 1.4 log10 copies of PCV2 per millilitre of sample (p<0.0001). This difference indicated that the assay from laboratory 1 yielded a higher output than the one from laboratory 2. Results also showed that there was no linear association between the amount of PCV2 DNA and the amount of total DNA, neither in nasal (p=0.86) nor in rectal (p=0.78) swabs, suggesting that normalizing of PCV2 DNA load in swab samples to total DNA concentration is not suitable. The present exploratory study highlights the need for the performance of ring trials on qPCV2 protocols between laboratories. Meanwhile, the proposed thresholds for PMWS diagnosis should only be considered reliable for each particular laboratory and each particular assay.     

The pharmacokinetics and metabolism of ivermectin in domestic animal species

The pharmacokinetic properties of drugs are closely related to their pharmacological efficacy. The kinetics of ivermectin are characterised, in general terms, by a slow absorption process, a broad distribution in the organism, low metabolism, and slow excretion. The kinetics vary according to the route of administration, formulation, animal species, body condition, age, and physiological status, all of which contribute to differences in drug efficacy. Characterisation of ivermectin kinetics can be used to predict and optimise the value of the parasiticide effects and to design programmes for parasite control. This article reviews the pharmacokinetics of ivermectin in several domestic animal species.     

Comparison between Haemophilus parasuis infection in colostrums-deprived and sow-reared piglets

The aim of this study was to compare the development of Glasser's disease in sow-reared and colostrum-deprived piglets. Ninety piglets from a commercial pig farm in Spain were used. The farm was positive for Haemophilus parasuis. Fifty-two pigs were sow-reared (SR) and 38 were colostrum-deprived (CD) piglets. The animals were intratracheally inoculated with H. parasuis serovar 5 and sacrificed at 1, 2 and 3 days post-infection. To assess the development of disease, antibody titers, clinical signs, pathological lesions, microbiological isolation and PCR amplification were compared between the groups. Inoculation of SR pigs did not cause clinical signs or lesions of Glasser's disease. In SR pigs, H. parasuis isolation and specific PCR amplification from tissues showed a very low number of positive samples. In contrast, in CD pigs, inoculation resulted in the typical signs and lesions of Glasser's disease. Positive microbiological isolation and specific PCR products were obtained from the majority of the tissues tested, and no antibodies against H. parasuis were detected. The experimental infection using CD pigs describes a successful method to study this microorganism and confirms the important role that maternal antibodies play in protection against clinical signs and disease.     

Adhesion of Escherichia coli K88ab fimbriae to porcine enterocytes: effect on enzymatic activities of the brush border, and cyclic AMP and GMP levels

We have studied the cellular alterations, after in vitro adherence, of purified K88ab fimbriae to membranes of porcine enterocytes. Effects on enzymatic activities as disaccharydases and alkaline phosphatase show low changes. While cAMP levels were decreased (44%), guanylyl cyclase was increased (up to 200%), and levels of cGMP were in consequence significantly affected. This study support the role of cyclic GMP as intracellular mediator for adherence, and suggest their implication in disease, affecting a membrane-mediated mechanisms for guanylate cyclase activation, that is unique in the intestine.     

Septicaemia secondary to infection by Corynebacterium macginleyi in an Indian python (Python molurus)

A seven-year-old female Indian python (Python molurus) weighing about 35kg was euthanased after several clinical episodes of stomatitis, pneumonia, ophthalmitis and dystocia over a period of four years. The animal had been maintained in a terrarium in a circus truck at an adequate temperature. During shows, however, the snake was considered to be exposed to stressful conditions for several hours at a time at low temperatures and with noise and bright lights. A post-mortem examination indicated ulcerative stomatitis, osteomyelitis, severe pneumonia and numerous granulomata and multifocal necrosis in stomach and spleen. Corynebacterium macginleyi was isolated in pure culture from the ulcerative stomatitis, and mixed with Stenotrophomonas maltophilia from the lungs and spleen. The findings indicated that the snake had died from a septicaemic process caused by C. macginleyi, probably originating from the stomatitis. The role of S. maltophilia as a secondary agent is discussed. The stress of the circus show and poor husbandry may have predisposed the animal to infection and septicaemia. This is the first report of C. macginleyi causing disease in a snake.     

Non-invasive assessment of growth, gender and time of day related changes of respiratory pattern in healthy cats by use of barometric whole body plethysmography

The objective of this study was to establish a reference base for respiratory variables (respiratory rate [R(R)], inspiratory and expiratory time [T(i) and T(e)], peak inspiratory and expiratory pseudoflow [PIF and PEF], tidal volume [V(T)], minute ventilation [V(E)] and enhanced pause [Penh]) of healthy cats by use of barometric whole body plethysmography (BWBP). Eighteen healthy European cats (10 male, 8 female) were studied from the age of 3 to 13 months in order to assess growth- and gender-related changes of BWBP variables. Chest radiographs and bronchoalveolar lavage cytology were performed to confirm pulmonary health status. Diurnal changes were investigated every 2 h over a period of 24 h when the cats were adult. V(T), V(E), PIF and PEF significantly increased during somatic growth and were higher in males than in females, whereas R(R), T(i), T(e), T(e)/T(i) ratio, PEF/PIF ratio and Penh remained unchanged and were not affected by gender. When measured over 24 h, Penh, T(e) and T(i) were significantly increased in the early morning hours (04:00 h), whereas R(R), PIF and PEF were decreased at that time. This study provides reference values of BWBP variables for healthy male and female cats and indicates when circadian changes might be observed.     

Effects of boar presence on agonistic behavior, shoulder scratches, and stress response of bred sows at mixing

The objective of this study was to determine the effects of boar presence on aggression, shoulder scratches, and salivary cortisol in group-housed sows during the period after mixing, which is when dominance hierarchies are formed. A total of 225 York-shire sows were used. Five groups of 15 sows were each exposed to 1 of 3 levels of boar presence (n = 15): physical (boar in pen with sows; 2.15 m2 per sow), fenceline (boar housed adjacent to sows; 2.3 m2 per sow), or control (no boar in the room; 2.3 m2 per sow). The experiment was divided into 2 phases. In phase 1, behavioral measurements were taken for 48 h after mixing and boar introduction. In phase 2, behavioral measurements were taken beginning on d 6 after mixing and included 24 h before and 48 h after boar removal. In phases 1 and 2, shoulder scratches were scored 24 h before and 24 h after mixing and boar removal, respectively. Saliva samples were collected each morning (0600 to 0700) and afternoon (1600 to 1700) during both phases. Frequencies of intersow aggressive contact (bite, body knock, and head knock) and threats as well as frequencies and durations of fighting bouts were determined from video recordings during daily feeding (49.6 +/- 1.4 min) and nonfeeding (21 +/- 0.2 h) periods. During phase 1, boar presence did not affect the frequency of threats during either the feeding or nonfeeding periods. Control groups had fewer aggressive contacts at feeding during the 25- to 48-h period compared with the fenceline groups (P = 0.01). Total duration of fighting and average fighting bout duration were unaffected by treatment. However, the number of fights occurring during feeding 25 to 48 h postmixing was lower in the physical groups than in the fenceline groups (P = 0.023); measures for the control groups were intermediate. Physical sows also received fewer shoulder scratches postmixing than did control sows (P = 0.048). After mixing, physical sows showed greater morning (P = 0.08) and afternoon (6 h postmixing, P = 0.01; 32 h postmixing, P = 0.08) salivary cortisol concentrations compared with fenceline sows, although they were only numerically greater than those for control sows. During phase 2, removing the boar did not increase fighting among sows or affect shoulder scratches or salivary cortisol concentrations. The presence of a boar was minimally effective at reducing fighting and scratches during the postmixing period, and sows showed a greater stress response in the presence of a boar.     

Use of protected methionine (Mepron M 85) in cattle

The ruminal stability of Mepron M 85 and the effect of supplementation with Mepron M 85 on free methionine level of blood were studied in rumen-fistulated cows and rumen- and duodenum-fistulated growing bulls. In five rumen-fistulated cows in situ 69.5% and 64.6% of the methionine content of Mepron M 85 was found after ruminal incubation of 16 h and 24 h, respectively. Daily rations of the rumen-fistulated cows were supplemented with 15.0 g DL-methionine and 17.7 g Mepron M 85, which increased the free methionine level of blood from 13.64 mumol/L to 15.35 and 20.46 mumol/L, respectively, three hours after feeding. In the four rumen- and duodenum-fistulated growing bulls, supplementation with 15.0 g DL-methionine and 17.7 g Mepron M 85 increased the total methionine getting into the duodenum during 24 h from 14.99 g to 16.84 and 20.84 g, respectively. The influence of Mepron M 85 on milk production was studied in 35 pairs of Hungarian Fleckvieh x Holstein-Friesian cows. The animals were coupled on the basis of the number of finished lactations, milk production in the previous lactation, and the date of calving. Daily supplementation of 18.0 g Mepron M 85 increased daily milk production significantly (p < 0.05), by 1.24 litres. Milk fat content also increased significantly (from 3.10% to 3.19%, p < 0.05) in the experimental group. The supplementation did not influence milk protein content.