EcPV2 DNA in equine genital squamous cell carcinomas and normal genital mucosa

Squamous cell carcinoma (SCC) represents the most common genital malignant tumor in horses. Similar to humans, papillomaviruses (PVs) have been proposed as etiological agents and recently Equine papillomavirus type 2 (EcPV2) has been identified in a subset of genital SCCs. The goals of this study were (1) to determine the prevalence of EcPV2 DNA in tissue samples from equine genital SCCs, penile intraepithelial neoplasia (PIN) and penile papillomas, using EcPV2-specific PCR, (2) to examine the prevalence of latent EcPV2 infection in healthy genital mucosa and (3) to determine genetic variability within EcPV2 and to disentangle phylogenetic relationships of EcPV2 among PVs. EcPV2 DNA was detected in all but one penile SCC (15/16), in all PIN lesions (8/8) and penile papillomas (4/4). Additionally, EcPV2 DNA was demonstrated in one of two metastasized lymph nodes, one contact metastasis in the mouth, two vaginal and one anal lesion. In healthy horses, EcPV2 DNA was detected in 10% (4/39) of penile swabs but in none of vulvovaginal swabs (0/20). This study confirms the presence of EcPV2 DNA in equine genital SCCs and shows its involvement in anal lesions, a lymph node and contact metastases. Latent EcPV2 presence was also shown in normal male genital mucosa. We found that different EcPV2 variants cocirculate among horses and that EcPV2 is related to the Delta+Zeta PVs and is only a very distant relative of high-risk human PVs causing genital cancer. Thus, similar viral tropism and similar malignant outcome of the infection do not imply close evolutionary relationship.     

MM3-ELISA evaluation of coproantigen release and serum antibody production in sheep experimentally infected with Fasciola hepatica and F. gigantica

During an experimental infection of sheep with Fasciola hepatica or F. gigantica, MM3-SERO and MM3-COPRO ELISA tests were applied to compare the kinetics of antibody production and coproantigen release between the 2nd and 32nd week post-infection (wpi). The Kato-Katz technique was used to measure the kinetics of egg shedding by both Fasciola species (eggs per gram of feces, epg). The kinetics of IgG antibodies for all sheep infected with F. hepatica and F. gigantica followed a similar pattern. Optical density (OD) increased rapidly between the 4th until the 12th wpi, when the highest values were reached and then decreased slowly until the 32nd wpi. Coproantigen levels increased above the cut-off value between 6 and 9 wpi in the F. hepatica group, and between 9 and 11 wpi in the F. gigantica group. The comparison between coproantigen levels and epg indicated that F. hepatica-infected sheep had detectable amounts of coproantigens 4–7 weeks before patency (egg shedding), while F. gigantica-infected sheep had detectable amounts of coproantigens 3–6 weeks before patency. When comparing the kinetics of coproantigen release vs the kinetics of epg, a similar pattern emerged, but with a two-week time-lag in epg, for both F. hepatica and F. gigantica infections. The amount of coproantigen release by each adult was not burden dependent for F. hepatica infection (burden of 33–66 adults), while it was for F. gigantica infection (burden of 17–69 adults). The results demonstrate the usefulness of the MM3-SERO and MM3-COPRO ELISAs as tools for the diagnosis of early as well as long-term fascioliasis infections, and suggest that they can potentially be applied to human fascioliasis even in countries where F. hepatica and F. gigantica co-exist. These tests can be employed not only in the diagnosis, but also in studies on epidemiology as well as pathogenesis and treatment in animals and humans since they allow post-treatment infection monitoring.     

Analgesic and motor-blocking action of epidurally administered levobupivacaine or bupivacaine in the conscious dog

Objective  To compare the analgesic and motor-blocking effects of epidurally administered levobupivacaine and bupivacaine in the conscious dog.
Study design  Prospective, randomized, cross-over study.
Animals  Six adult female Beagle dogs.
Methods  Each animal received three doses of levobupivacaine or bupivacaine (0.5, 1.0 and 1.5 mg kg⁻¹; concentrations 0.25%, 0.50%, and 0.75%, respectively) in a total volume of 0.2 mL kg⁻¹ by means of a chronically implanted epidural catheter. Onset, duration (through pinch response in the sacral, lumbar and toe areas) and degree of analgesia and motor-blocking status was determined with a scoring system and at regular intervals over 8.5 hours before (baseline) and after drug administration.
Results  Epidurally administered levobupivacaine and bupivacaine had a similar dose-dependent analgesic action with no significant differences in onset (range: 5–8 minutes), duration (bupivacaine: 42 ± 28, 135 ± 68 and 265 ± 68 minutes, and levobupivacaine: 28 ± 33, 79 ± 55 and 292 ± 133 minutes; 0.25%, 0.50%, and 0.75%, respectively) or maximum degree of analgesia. However, levobupivacaine tended to produce a shorter duration of motor block than bupivacaine and the difference in the motor to nociceptive blockade times was significant at the highest dose.
Conclusion  Epidural levobupivacaine produced an analgesic action similar to that of bupivacaine.
Clinical relevance  Epidural levobupivacaine is suitable for clinical use in dogs, mostly at the highest dose if a high degree of analgesia is required.     

Effects of Stocking Rate and Heather Supplementation on Gastrointestinal Nematode Infections and Host Performance in Naturally-Infected Cashmere Goats

The aim of this study, performed on 62 adult dry cashmere goats grazing upland perennial ryegrass–white clover pastures and naturally infected with gastrointestinal nematodes, was to investigate the effects of stocking rate (SR: 24 vs. 38 goats · ha^-1) and tannin-containing heather supplementation (H: Calluna vulgaris [L.] Hull, Erica spp.) vs. nonsupplementation on parasite burden, fecal egg counts (FEC), and live weight (LW) changes. Goats were randomly assigned to four treatments in a 2 × 2 factorial arrangement and grazed continuously from May to October. Six goats per treatment were slaughtered at the end of the grazing period, and adult worms in the abomasum and small and large intestines of each animal were recovered, counted, and identified. FEC was affected by SR (P < 0.01) but not by H. However, the SR × H interaction was significant (P < 0.05). FEC increased (P < 0.001) along the grazing season in all treatments, and the SR × time interaction was significant (P < 0.001). In general, mean total worm counts in abomasum and small intestine tended to be higher under high SR, although the differences were only significant (P < 0.01) in Trichostrongylus spp. counts. In goats managed under the high SR, the mean of total Teladorsagia circumcincta counts was lower (P < 0.01) in supplemented animals, but no differences were recorded for Trichostrongylus spp., Chabertia ovina, Oesophagostomum columbianum, and Trichuris ovis. The goats gained more LW (P < 0.001) under low SR and when they were heather-supplemented. No significant SR × H interaction was found for LW change. In conclusion, high stocking rate increases the infectivity risk of pasture and the supplementation of grazing goats with heather contributing to improve animals' performance. Notwithstanding, the effect of heather availability on nematode FEC reduction could be highly dependent on the climatic conditions.     

Polynomial order selection in random regression models via penalizing adaptively the likelihood

Orthogonal Legendre polynomials ( LP ) are used to model the shape of additive genetic and permanent environmental effects in random regression models ( RRM ). Frequently, the Akaike ( AIC ) and the Bayesian ( BIC ) information criteria are employed to select LP order. However, it has been theoretically shown that neither AIC nor BIC is simultaneously optimal in terms of consistency and efficiency. Thus, the goal was to introduce a method, ‘penalizing adaptively the likelihood’ ( PAL ), as a criterion to select LP order in RRM. Four simulated data sets and real data (60 513 records, 6675 Colombian Holstein cows) were employed. Nested models were fitted to the data, and AIC, BIC and PAL were calculated for all of them. Results showed that PAL and BIC identified with probability of one the true LP order for the additive genetic and permanent environmental effects, but AIC tended to favour over parameterized models. Conversely, when the true model was unknown, PAL selected the best model with higher probability than AIC. In the latter case, BIC never favoured the best model. To summarize, PAL selected a correct model order regardless of whether the ‘true’ model was within the set of candidates.     

Constraints on dairy cattle productivity at the smallholder level in the Philippines

Survey data on dairy cattle production were gathered in two sites [Site I (three-year survey) and Site II (two-year survey)] in Southern Luzon, Philippines. Crossbred (Holstein-Friesian x Sahiwal) dairy cows (n = 122) managed by smallholder farmers belonging to five primary cooperatives under the federation of dairy farmers, were monitored monthly for milk production, feed intake and availability, and reproduction and health status. The purpose of the survey was to identify constraints to productivity. The reproductive status of the cows was monitored by measuring milk and plasma progesterone concentrations by radioimmunoassay and rectal palpation of the ovaries. Plasma concentrations of selected metabolites [beta-hydroxybutyrate (BHB), inorganic phosphorus, albumin, globulin, urea] were also measured at one month before calving and at one month and 2-3 months postpartum, to determine if these could serve as biochemical indicators of nutritional stress. A long calving interval (CI = > 400 days) was identified as the major constraint to productivity of dairy cattle on smallholder farms. The three main problems related to this reproductive constraint were: (1) poor breeding management, in particular lack of accurate estrus detection; (2) repeat breeding, i.e. three or more services were required before conception; and (3) poor ovarian function, shown by some cows with lose progesterone levels. An important cause of these problems was undernutrition, particularly at critical periods of the cow's reproductive life, reflected in the slow recovery from loss in body weight and condition score during the early postpartum period and the increased plasma BHB values at peripartum period in some cows, indicative of negative energy balance, and the flat lactation profile. These findings are useful and relevant as a database in the development of an appropriate management scheme aimed toward improving dairy cattle production and productivity at smallholder level. It highlights the importance of estrus detection, good breeding management and the use of a practical strategic nutritional supplementation, particularly during stressful periods in the cows's reproductive life.     

Comparison between meloxicam and transdermally administered fentanyl for treatment of postoperative pain in dogs undergoing osteotomy of the tibia and fibula and placement of a uniplanar external distraction device

OBJECTIVE: To compare the efficacy of meloxicam administered perioperatively with transdermal administration of fentanyl via a patch placed preoperatively in dogs undergoing orthopedic surgery. DESIGN: Prospective study. ANIMALS: 16 dogs. PROCEDURE: Unilateral or bilateral osteotomy of the tibia and fibula was surgically performed, and a uniplanar external distraction device was placed in each limb. Postoperative pain and lameness were assessed 24, 48, and 72 hours after administration of the first of 3 doses of meloxicam (0.2 mg/kg [0.09 mg/lb], IV, given preoperatively, followed by 0.1 mg/kg [0.045 mg/lb], IV, after 24 hours, and 0.1 mg/kg, PO, after 48 hours) or preoperative placement of a transdermal fentanyl patch (50 microg/h) left in place for 72 hours. RESULTS: No significant differences in total pain scores were detected between groups. Mean +/- SD lameness scores assessed at 24 and 72 hours were lower in dogs in the meloxicam group than dogs in the fentanyl group. Lameness scores decreased with time in a similar manner in both treatment groups. CONCLUSIONS AND CLINICAL RELEVANCE: Perioperative administration of meloxicam or preoperative placement of a transdermal fentanyl patch provided effective and similar postoperative analgesia in dogs undergoing orthopedic surgery. However, because of its anti-inflammatory effects, treatment with meloxicam reduced the degree of lameness and resulted in rapid functional recovery of the limb.     

Development of a genetic manipulation system for Haemophilus parasuis

Haemophilus parasuis is a member of the family Pasteurellaceae and an important respiratory-tract pathogen of swine, which is the etiological agent of Glasser's disease. Because no genetic manipulation system is available for H. parasuis so far, in vivo studies about the role of its genes involved in virulence are unfeasible. Here we demonstrate that H. parasuis has a cyclic AMP (cAMP)-dependent natural transformation system that enables the uptake of DNA in which the ACCGAACTC sequence signal must be present. After improving DNA transformation parameters, such as cAMP and DNA concentration and exposition time of the exogenous DNA, a knockout mutant of H. parasuis defective in the thy gene, encoding the thymidylate synthase enzyme, has been constructed. Data presented in this work open the possibility for the functional analysis of genes involved in the infectious process of this animal pathogen.