Safety of an attenuated West Nile virus vaccine, live Flavivirus chimera in horses

REASON FOR PERFORMING STUDY: West Nile virus (WNV) infection is endemic and able to cause disease in naive hosts. It is necessary therefore to evaluate the safety of new vaccines. OBJECTIVES: To establish: 1) the safety of a modified live Flavivirus/West Nile virus (WN-FV) chimera by administration of an overdose and testing for shed of vaccine virus and spread to uninoculated sentinel horses; 2) that this vaccine did not become pathogenic once passaged in horses; and 3) vaccine safety under field conditions. METHODS: There were 3 protocols: 1) In the overdose/shed and spread study, horses were vaccinated with a 100x immunogenicity overdose of WN-FV chimera vaccine and housed with sentinel horses. 2) A reversion to virulence study, where horses were vaccinated with a 20x immunogenicity overdose of WN-FV chimera vaccine. Horses in both studies were evaluated for abnormal health conditions and samples obtained to detect virus, seroconversion and dissemination into tissues. 3) In a field safety test 919 healthy horses of various ages, breeds and sex were used. RESULTS: Vaccination did not result in site or systemic reactions in either experimental or field-injected horses. There was no shed of vaccine virus, no detection of vaccine virus into tissue and no reversion to virulence with passage. CONCLUSIONS: WN-FV chimera vaccine is safe to use in horses with no evidence of ill effects from very high doses of vaccine. There was no evidence of reversion to virulence. In addition, administration of this vaccine to several hundred horses that may have been previously exposed to WNV or WNV vaccine resulted in no untoward reactions. POTENTIAL RELEVANCE: These studies establish that this live attenuated Flavivirus chimera is safe to use for immunoprophylaxis against WNV disease in horses.     

Outcomes after extrahepatic portosystemic shunt ligation in 49 dogs

OBJECTIVE: To evaluate outcomes after attenuation of extrahepatic portosystemic shunts in dogs using surgical silk. DESIGN: Retrospective study. PROCEDURE: Case records were reviewed for degree of surgical attenuation, experience of the primary surgeon, perioperative mortality and problems related to persistent portosystemic shunting or shunt ligation. Presence of portosystemic shunting after surgery was evaluated by ammonia tolerance testing, measurement of postprandial serum bile acid, plasma urea and cholesterol concentrations and liver enzyme activity. The influence of age, postocclusion portal pressure, primary surgeon, degree of attenuation and postoperative biochemical findings on the occurrence of postoperative problems was assessed. RESULTS: The mortality rate was 2.1%. Shunt attenuation was complete in 34% and partial in 66% of dogs. Portal hypertension necessitating ligature removal was encountered in only one dog. Five dogs experienced neurological abnormalities (seizures or ataxia), possibly as a manifestation of 'postligation seizure syndrome'. Postoperative liver function was normal in 78% of dogs, including 70% with partial shunt attenuation. Experience of the surgeon was related positively to outcome after partial attenuation (P = 0.002). Postoperative biochemical evidence of abnormal liver function was the most sensitive predictor of recurrence of clinical signs referable to persistent portosystemic shunting. CONCLUSIONS: In the hands of an experienced surgeon, surgical attenuation of single extrahepatic shunts was safe and effective, even in animals with partial attenuation. Most dogs with biochemical evidence of persistent shunting suffer relapse of clinical signs within 18 months of surgery. Postligation neurological syndromes of variable intensity may be more common than previously thought.     

Effect of oxytocin on expression of cytosolic phospholipase A2 mRNA and protein in ovine endometrial tissue in vivo

The induction of endometrial prostaglandin (PG) F₂ synthesis by oxytocin is dependent upon activation of phospholipase (PL) A₂ and mobilization of arachidonic acid. The objective of this study was to determine if oxytocin stimulates PGF₂ synthesis by inducing synthesis of cytosolic PLA₂ (cPLA₂). In Experiment 1, 15 ovariectomized ewes were given progesterone and estradiol to simulate an estrous cycle. Ewes were then given an injection of oxytocin on Day 14 of the simulated estrous cycle. Jugular blood samples were collected and assayed for 13,14-dihydro-15-keto-prostaglandin F₂ (PGFM). Uteri were collected at 0, 7.5, 25, 90, or 240 min postinjection (n = 3 ewes/time point). Total RNA was isolated from caruncular endometrium and subjected to dot-blot analysis. Oxytocin induced a rapid and transient increase in serum PGFM (P < 0.01). However, endometrial concentrations of cPLA2 mRNA did not change following oxytocin administration (P > 0.10). In Experiment 2, 11 ovary-intact ewes were given oxytocin (n = 5) or saline (n = 6) on Day 15 after estrus. Jugular blood samples were collected and assayed for serum concentrations of PGFM. Uteri were collected at 15 min postinjection. Homogenates were prepared from caruncular endometrium and subjected to Western blot analysis. Concentrations of PGFM were higher in oxytocin treated ewes compared to saline treated ewes at 15 min postinjection (P < 0.01). Endometrial concentrations of cPLA₂ protein were greater in the cytosolic than in the microsomal fraction (P < 0.01). Oxytocin did not affect the amount of cPLA₂ protein in either fraction (P > 0.10). In conclusion, oxytocin did not effect expression of either cPLA₂ mRNA or protein in ovine endometrium. Oxytocin may stimulate PGF₂ synthesis by activating cPLA₂ protein that is already present in an inactive form.     

Technical note: Constrained optimization of breed composition in composite populations to balance net merit and risk

We developed a method to derive optimum composite genotypes when the proportion of a specific breed (or combination of breeds) in the composite is constrained to a desired level. The method allows development of composites that balance net merit according to available parameters and reflects the production system and(or) risk preference, particularly when traits in the breeding objective do not represent all traits important for production. The method can also be used to evaluate the cost of constraining breed use. For example, when the true optimal composite is difficult to obtain in the time frame available to the producer, the predicted merit of the composite with breed proportion restricted to those that are obtainable in practice can be compared with the predicted merit of the true optimum composite. In a case study the method is used to optimize a tropical composite using Brahman, Charolais, and Belmont Red breeds. The proportion of Charolais is to be constrained to 50%. Using growth parameters from a tropical crossbreeding experiment, the unconstrained optimum composite was 32.5% Brahman, 11.1% Belmont Red, and 56.3% Charolais. When the proportion of Charolais in the composite was constrained to 50%, optimum proportions of Brahman and Belmont Red were 33.9% and 16.1% respectively. In the case study net merit (annualized profit per cow exposed, $), as defined by the growth parameters, was .2% less than that of the unconstrained optimum composite.     

Low concentrations of zearalenone in diets of boars for a prolonged period of time

Thirty-two boars were fed diets containing 0, 3, 6 or 9 ppm zearalenone from 32 d of age up to 145 or 312 d of age. Rate of gain, feed intake and gain:feed were not significantly different among treatments. Feeding of diets containing 9 ppm zearalenone did not affect the libido in seven boars. There was a tendency for boars fed 9 ppm dietary zearalenone to produce lower total and gel free volumes of semen, with lower total motile sperm; however, only the percent motility of the sperm was reduced (P less than .05).     

COMPARISON OF ULTRASONOGRAPHIC AND RADIOGRAPHIC MEASUREMENTS OF CANINE PROSTATE DIMENSIONS

The purposes of this study were to: 1) compare prostatic dimensions measured on radiographic and ultrasonographic images and 2) compare a subjective radiographic assessment of prostate size with a previously-described objective method. Thirty-four male dogs undergoing investigation of prostatic disorders were used. Prostate length and depth were measured from ultrasonographic and radiographic images. A subjective assessment of prostate size ('small', 'normal', or 'enlarged') was made in 29 animals by one of the authors who was unaware of radiographic or ultrasonographic measurements. In addition, the distance from sacral promontory to the pubic brim was also measured. A prostate length or depth of >70% of this distance was defined as 'enlarged' and <70% as 'normal'. After the effects of magnification on radiographic measurements were eliminated, there were no significant differences between prostatic length measured by the two methods. However, a significant difference was obtained between prostatic depth measurements. The subjective assessment agreed with a previously described objective assessment of prostatic size in 21/29 dogs for prostate length but in only 12/29 for depth. Prostatic length varied from 46.6 to 116.4% (mean 75.7%) of the distance from the pubic brim to the sacral promontory. Prostatic depth varied from 33.0% to 94.6% (mean 59.7%) of the same distance. It is recommended that prostate length, rather than depth, be used when evaluating prostate size from lateral abdominal radiographs.     

Vertical distribution, size and composition of the weed seedbank under various tillage and herbicide treatments in a sequence of industrial crops

Investigations were conducted during the 2003, 2004 and 2005 growing seasons in northern Greece to evaluate effects of tillage regime (mouldboard plough, chisel plough and rotary tiller), cropping sequence (continuous cotton, cotton–sugar beet rotation and continuous tobacco) and herbicide treatment on weed seedbank dynamics. Amaranthus spp. and Portulaca oleracea were the most abundant species, ranging from 76% to 89% of total weed seeds found in 0–15 and 15–30 cm soil depths during the 3 years. With the mouldboard plough, 48% and 52% of the weed seedbank was found in the 0–15 and 15–30 cm soil horizons, while approximately 60% was concentrated in the upper 15 cm soil horizon for chisel plough and rotary tillage. Mouldboard ploughing significantly buried more Echinochloa crus‐galli seeds in the 15–30 cm soil horizon compared with the other tillage regimes. Total seedbank (0–30 cm) of P. oleracea was significantly reduced in cotton–sugar beet rotation compared with cotton and tobacco monocultures, while the opposite occurred for E. crus‐galli. Total seed densities of most annual broad‐leaved weed species (Amaranthus spp., P. oleracea, Solanum nigrum) and E. crus‐galli were lower in herbicide treated than in untreated plots. The results suggest that in light textured soils, conventional tillage with herbicide use gradually reduces seed density of small seeded weed species in the top 15 cm over several years. In contrast, crop rotation with the early established sugar beet favours spring‐germinating grass weed species, but also prevents establishment of summer‐germinating weed species by the early developing crop canopy.     

The vitamin A requirement and the vitamin A status of growing cattle. 1. Studies of calves

Five experiments with 18 to 36 male calves each of the black and white dairy cattle breed (age: 14-21 days, initial live weight: approximately 45 kg per animal) were carried out in order to investigate the influence of various vitamin A supply (0-80,000 IU per 100 kg LW and day) on dry matter intake and weight gain as well as the vitamin A status of liver and blood plasma over 84 days. The calves consumed a diet free of carotene and vitamin A consisting of milk replacer, concentrate and chopped wheat straw. The calves were fed in three experiments for a longer time in order to observe the further vitamin A depletion. Nine animals consumed an unsupplemented ration, nine other one got 10,000 IU vitamin A per 100 kg LW and day. Biopsies of liver and plasma samples were taken from 4 animals per group every four weeks. The various vitamin A supplementation did not significantly influence the dry matter intake (Mean: 1.67; 1.48 to 1.80 kg DM per animal and day) and the weight gain of calves (Mean: 702, 599 to 770 g per animal and day). First vitamin A deficiency symptoms (reduced feed intake, decreased weight gain, diarrhoea etc.) were observed in animals of unsupplemented group after 100 days of experiments. After 84 days the vitamin A concentration of liver of animals of unsupplemented groups decreased to 1.3-32.2% compared with the begin of experiments (60.6-155.7 mumol/kg fresh matter). Up to 51% of initial concentration were found when 10,000 IU vitamin A per 100 kg LW and day were fed. About 25,000 IU vitamin A per 100 kg LW and day were required in order to keep the initial level of vitamin A concentration of liver. The plasma vitamin A concentration is unsuitable for estimation of vitamin A status of calves. The concentration of vitamin A of liver and plasma amounted to 114 mumol per kg and 0.25 mumol per litre at the begin of experiments. The vitamin A concentration of liver of unsupplemented group decreased to 20 mumol per kg, that of plasma increased to 0.28 mumol per 1 at the end. A strong vitamin A deficiency (liver concentration: less than 10 mumol/kg) may cause a decrease of vitamin A concentration of blood.     

Simultaneous intramammary and intranasal inoculation of lactating cows with bovine herpesvirus 4 induce subclinical mastitis

In this study, we examined whether an experimental bovine herpesvirus 4 (BHV4) infection can induce bovine mastitis, or can enhance bovine mastitis induced by Streptococcus uberis (S. uberis). Four lactating cows were inoculated intramammarily and intranasally with BHV4, and four lactating control cows were mock-inoculated. After 14 days, two of four cows from each group were inoculated intramammarily with S. uberis. No clinical signs were recorded in cows inoculated only with BHV4, and their milk samples showed no abnormal morphology, despite the fact that BHV4 replicated in inoculated quarters. Somatic cell count increased significantly in milk from three of six BHV4-inoculated quarters, compared to the non-inoculated quarters of the same cows (within-cow) and the quarters of mock-inoculated cows (control group) on days 8, 9 and 11 post-inoculation (pi). BHV4 was isolated from nasal swabs between days 2 and 9 pi. Clinical mastitis was observed in all four cows intramammarily inoculated with S. uberis. A preceding BHV4 infection did not exacerbate the clinical mastitis induced by S. uberis. S. uberis infections appeared to trigger BHV4 replication. From one quarter of each of two cows inoculated with BHV4 and S. uberis, BHV4 was isolated, and not from quarters inoculated with BHV4 only. In conclusion, BHV4 did not induce bovine clinical mastitis after simultaneous intranasal and intramammary inoculation. However, the BHV4 infection did induce subclinical mastitis in 50% of the cows and the quarters.