The NS3 proteins of global strains of bluetongue virus evolve into regional topotypes through negative (purifying) selection

Comparison of the deduced amino acid sequences of the genes (S10) encoding the NS3 protein of 137 strains of bluetongue virus (BTV) from Africa, the Americas, Asia, Australia and the Mediterranean Basin showed limited variation. Common to all NS3 sequences were potential glycosylation sites at amino acid residues 63 and 150 and a cysteine at residue 137, whereas a cysteine at residue 181 was not conserved. The PPXY and PS/TAP late-domain motifs were conserved in all but three of the viruses. Phylogenetic analyses of these same sequences yielded two principal clades that grouped the viruses irrespective of their serotype or year of isolation (1900-2003). All viruses from Asia and Australia were grouped in one clade, whereas those from the other regions were present in both clades. Each clade segregated into distinct subclades that included viruses from single or multiple regions, and the S10 genes of some field viruses were identical to those of live-attenuated BTV vaccines. There was no evidence of positive selection on the S10 gene as assessed by reconstruction of ancestral codon states on the phylogeny, rather the functional constraints of the NS3 protein are expressed through substantial negative (purifying) selection.     

Comparison of protocols to synchronize estrus and ovulation in estrous-cycling and prepubertal beef heifers

The objective of the experiment was to compare follicular dynamics, ovulatory response to GnRH, and synchrony of estrus and ovulation among estrous-cycling and prepubertal beef heifers synchronized with a controlled internal drug-release (CIDR)- based or GnRH-PGF(2alpha) (PG) protocol. Estrous-cycling beef heifers were randomly assigned to 1 of 4 treatments (C1, C2, C3, C4), and prepubertal beef heifers were randomly assigned to 1 of 2 treatments (P1, P2) by age and BW. Blood samples were taken 10 and 1 d before treatment to confirm estrous cyclicity status (progesterone > or =0.5 ng/mL estrous cycling). The CIDR Select (C1, n = 12; P1, n = 14)-treated heifers received a CIDR insert (1.38 g of progesterone) from d 0 to 14, GnRH (100 microg, i.m.) on d 23, and PG (25 mg, i.m.) on d 30. Select Synch + CIDR (C2, n = 12; P2, n = 11)-treated heifers received a CIDR insert and GnRH on d 23 and PG at CIDR removal on d 30. The CIDR-PG (C3, n = 12)-treated heifers received a CIDR insert on d 23 and PG at CIDR removal on d 30. Select Synch (C4, n = 12)-treated heifers received GnRH on d 23 and PG on d 30. HeatWatch transmitters were fitted at CIDR removal (C1, C2, C3, P1, and P2) or at GnRH administration (C4) for estrus detection. Ultrasound was used to determine the response to GnRH and the timing of ovulation after estrus. Among the estrous-cycling heifers, ovulatory response to GnRH and estrous response did not differ (P > 0.05). Among the prepubertal heifers, more (P = 0.02) P1 heifers responded to GnRH than P2 heifers, but estrous response did not differ (P > 0.05). Among the estrous-cycling heifers, variance for interval to estrus after PG was reduced (P < 0.05) for C1 compared with each of the other treatments, and C3 [corrected] was reduced (P < 0.05) compared with C2 [corrected] Variance for interval to ovulation after PG was reduced (P < 0.05) for C1 compared with each of the other treatments. Among the prepubertal heifers, there was no difference (P > 0.05) in variance for interval to estrus or ovulation. Results from C1 and P1 (T1) and C2 and P2 (T2) were combined to compare T1 and T2 among mixed groups of estrous-cycling and prepubertal heifers. Response to GnRH was greater (P < 0.01; 81% T1 and 39% T2), and variances for interval to estrus and ovulation for T1 were reduced (P < 0.01) compared with T2. In summary, CIDR Select improved (P < 0.01) the synchrony of estrus and ovulation compared with Select Synch + CIDR.     

Report of the 3rd Havemeyer workshop on allergic diseases of the Horse, Hólar, Iceland, June 2007

Allergic diseases occur in most mammals, although some species such as humans, dogs and horses seem to be more prone to develop allergies than others. In horses, insect bite hypersensitivity (IBH), an allergic dermatitis caused by bites of midges, and recurrent airway obstruction (RAO), a hyperreactivity to stable born dust and allergens, are the two most prevalent allergic diseases. Allergic diseases involve the interaction of three major factors: (i) genetic constitution, (ii) exposure to allergens, and (iii) a dysregulation of the immune response determined by (i) and (ii). However, other environmental factors such as infectious diseases, contact with endotoxin and degree of infestation with endoparasites have been shown to influence the prevalence of allergic diseases in humans. How these factors may impact upon allergic disease in the horse is unknown at this time. The 3rd workshop on Allergic Diseases of the Horse, with major sponsorship from the Havemeyer Foundation, was held in Hólar, Iceland, in June 2007 and focussed on immunological and genetic aspects of IBH and RAO. This particular venue was chosen because of the prevalence of IBH in exported Icelandic horses. The incidence of IBH is significantly different between Icelandic horses born in Europe or North America and those born in Iceland and exported as adults. Although the genetic factors and allergens are the same, exported adult horses show a greater incidence of IBH. This suggests that environmental or epigenetic factors may contribute to this response. This report summarizes the present state of knowledge and summarizes important issues discussed at the workshop.     

Leptospirosa pomona Abortion Storm in a Cattle Herd in Saskatchewan

Abortions occurred in 18% of 131 beef cows and heifers during two months, on a farm in southern Saskatchewan. The losses began two weeks after acute febrile illness and agalactia in a dairy cow to which the beef herd had been exposed. A diagnosis of Leptospira interrogans serovar pomona infection was made on the basis of serology in cows and the finding of leptospires in fetal tissues by fluorescent antibody test. Tentative diagnosis of infectious bovine rhinotracheitis delayed treatment and prophylaxis until infection attained high intensity in the herd and severe losses to the farmer occurred. Abortions ceased after vaccination against pomona and oxytetracycline treatment of pregnant cows, although chronic debility followed the acute phase of the disease in some cows. Recrudescence of infection was suspected four months later, when acute agalactia occurred in one cow and debility in calves and cows was recurring. Pomona infection was not proven, but dihydrostreptomycin treatment and revaccination were applied to the whole herd. Seroconversion and IgM antibody continued to indicate a persistent source of infection and susceptibility in a minority of the population one year after onset. The source of the original infection is believed to have been a carrier beef cow, or a dairy cow which was leptospiruric at the time of contact with the beef herd. With the exception of one aborted calf, no evidence of pomona infection was found outside the farm, in cattle or wild mammals tested serologically within a radius of 30 km, during one year following the outbreak.     

Idiopathic granulomatous pneumonia in seven horses

The history, clinical signs and pathological findings in seven adult horses with histologically confirmed idiopathic granulomatous disease, primarily of the lungs, are reviewed. They ranged in age from eight to 21 years, five were geldings and two were females, they belonged to five breeds and there were no seasonal or geographical associations. The primary clinical signs were chronic weight loss, exercise intolerance and respiratory distress which did not respond to conventional treatment. The most consistent physical findings were depression, anorexia, tachycardia, tachypnoea and adventitious lung sounds. Thoracic radiographs revealed a diffuse, structured, nodular, interstitial pulmonary pattern in each horse. Haematological measurements suggested a chronic inflammatory process and the cytology of transtracheal washes was consistent with a mild suppurative inflammation. Idiopathic granulomatous pneumonia was confirmed histologically in each of the horses, either postmortem or by a lung biopsy. The horses responded poorly to medical treatment and only one of three treated horses is still alive.     

Pharmacokinetics of firocoxib after intravenous administration of multiple consecutive doses in neonatal foals

The purpose of this study was to determine the pharmacokinetic profile of intravenous firocoxib in neonatal foals. Six healthy foals were administered 0.09 mg/kg firocoxib intravenously once a day for 7 days. Blood was collected for plasma firocoxib analysis using high‐performance liquid chromatography with fluorescence detection at times 0 (day 1 of study only) and 0.08, 0.25, 1, 2, 4, 6, 8, 16 and 24 hr on dose numbers 1, 5 and 7. Blood was also collected immediately prior to doses 3, 4, 5 and 7. Final samples were collected at 36, 48, 72 and 96 hr following the final dose. Noncompartmental analysis using the trapezoidal method with linear interpolation revealed a moderate half‐life (15.9 ± 9.1 hr) with a large volume of distribution at steady state (1.79 ± 0.57 L/kg) and a clearance (96.0 ± 59.2 ml h^?1 kg^?1) that was more rapid than that observed in adult horses.     

Predicting percentage of intramuscular fat using two types of real-time ultrasound equipment

In the present study, 500 steers were used to develop models for predicting the percentage of intramuscular fat (PIMF) in live beef cattle. Before slaughter, steers were scanned across the 11th and 13th ribs using Aloka 500V (AL-500) and Classic Scanner 200 (CS-200) machines. Four to five images were collected per individual steer using each machine. After slaughter, a cross-sectional slice of the longissimus muscle from the 12th rib facing was used for chemical extraction to determine actual carcass percentage of intramuscular fat (CPIMF). Texture analysis software was used by two interpreters to select a region for determination of image parameters, which included Fourier, gradient, histogram, and co-occurrence parameters. Four prediction models were developed separately for each of AL-500 and CS-200 based on images captured by the respective machines. These included models developed without transformation of CPIMF (Model I), models based on logarithmic transformation of CPIMF (Model II), ridge regression procedure (Model III), and principal component regression procedure (Model IV). Model R2 and root mean square error of AL-500 Models I, II, III, and IV were 0.72, 0.84%; 0.72, 0.85%; 0.69, 0.91%; and 0.71, 0.86%; respectively. The corresponding R2 and root mean square error values of CS-200 Models I, II, III, and IV were 0.68, 0.87%; 0.70, 0.85%; 0.64, 0.94%; and 0.65, 0.91%; respectively. Initially, AL-500 and CS-200 prediction models were validated separately on an independent data set from 71 feedlot steers. The overall mean bias, standard error of prediction, and rank correlation coefficient across the four AL-500 models were 0.42%, 0.84%, and 0.88, respectively. For the four CS-200 models, the corresponding overall mean values were 0.67%, 0.81%, and 0.91, respectively. In a second validation test, only Model II of AL-500 and CS-200 was evaluated separately based on data from 24 feedlot steers. The overall mean bias, absolute difference, and standard error of prediction of AL-500 Model II were 0.71, 0.92, and 0.98%. For CS-200 Model II, the corresponding values were 0.59, 0.97, and 1.03%. Both AL-500 and CS-200 equipment can be used to accurately predict PIMF in live cattle. Further improvement in the accuracy of prediction equations could be achieved through increasing the development data set and the variation in PIMF of cattle used.     

Laparoscopic ovariectomy using sequential electrocoagulation and sharp transection of the equine mesovarium

OBJECTIVE: To describe in horses and ponies a laparoscopic ovariectomy technique facilitated by electrosurgical instrumentation. STUDY DESIGN: Elective ovariectomy was performed in 23 mares using laparoscopic electrosurgical instrumentation. ANIMALS OR SAMPLE POPULATION: Twenty-three mares (13 horses, 10 ponies), aged from 2 to 21 years and weighing 90 to 545 kg. METHODS: Food was withheld for a minimum of 12 hours. Mares were sedated with detomidine hydrochloride (0.02 to 0.03 mg/kg) or xylazine hydrochloride (0.5 to 1.0 mg/kg). Excluding the pony mares, all other mares were restrained in stocks. Portal sites in the paralumbar fossa region were desensitized with 2% mepivacaine. Abdominal insufflation was achieved through a teat cannula positioned in the ventral abdomen or a Verres-type needle placed through the paralumbar fossa. After trocar and laparoscope insertion, the ipsilateral ovary and mesovarium were identified, and the mesovarium, tubal membrane, and proper ligament were infiltrated with 2% mepivacaine. The mesovarium was coagulated using bipolar or monopolar electrosurgical forceps and transected sequentially from cranial to caudal until the ovary was completely freed and then removed. The contralateral ovary was removed in a similar fashion through the opposite paralumbar fossa. RESULTS: Bipolar and monopolar electrosurgical forceps were easy to use and provided adequate coagulation of vessels within the mesovarium. Two mares were euthanatized after the procedure for unrelated reasons. One mare had mild signs of colic 24 hours after ovariectomy. In 1 pony mare, the incision used to remove one ovary dehisced on the 5th postoperative day and was allowed to heal by second-intention. No long-term complications had occurred in 11 horses and 10 ponies, 6 to 24 months after surgery. CONCLUSION: Laparoscopic ovariectomy and hemostasis of the mesovarium can be easily accomplished using electrosurgical instrumentation. CLINICAL RELEVANCE: Standing laparoscopic ovariectomy, using electrosurgical instrumentation, is an effective and safe technique to provide hemostasis of the mesovarium in mares.     

A review of tissue reference values used to assess the trace element status of farmed red deer (Cervus elaphus)

AIMS: This paper reviews the principles for the establishment of biochemical reference criteria for assessing the trace element status of farmed livestock and summarises data for copper, selenium, vitamin B12 and iodine for farmed red deer. COPPER: Enzootic ataxia and osteochondrosis occur when liver copper concentrations are below 60 micromol/kg fresh tissue, and serum copper concentrations are below 3-4 micromol/l. Growth responses to copper supplementation have been equivocal when blood copper concentrations were 3-4 micromol/l, but were significant when mean blood copper concentrations were 0.9-4.0 micromol/l. No antler growth or bodyweight response to copper supplementation was observed when blood ferroxidase levels averaged 10-23 IU/l (equivalent to serum copper concentrations of 6-13 micromol/l) and liver copper concentrations averaged 98 mumol/kg fresh tissue. These data suggest that 'deficient', 'marginal' and 'adequate' ranges for serum copper concentrations should be 5, 5-8, and 8 micromol/l, respectively, and those for liver copper concentrations should be 60, 60-100, and 100 micromol/kg, respectively. SELENIUM: White muscle disease has been reported in young deer with blood and liver selenium concentrations of 84-140 nmol/l and 240-500 nmol/kg fresh tissue, respectively. No growth-rate response to selenium supplementation occurred in rising 1-year-old deer when blood selenium concentrations were less than 130 nmol/l, the range in which a growth-rate response would be expected in sheep. VITAMIN B12: Vitamin B12 concentrations in deer are frequently below 185 pmol/l without clinical or subclinical effects. No growth response was observed in young deer with vitamin B12 concentrations as low as 75-83 pmol/l. A growth response to cobalt/vitamin B12 supplementation occurs in lambs with serum vitamin B12 concentrations 336 pmol/l. CONCLUSIONS: Data that can be used to establish reference ranges for assessing trace element status in deer are limited. More robust reference values for farmed red deer need to be established through further studies relating biochemical data to health and performance.