Validation of betapropiolactone (BPL) as an inactivant for infectious bovine rhinotracheitis (IBR) virus

Infectious bovine rhinotracheitis (IBR) virus causes vulvovaginitis, abortion and respiratory disease in cows and heifers. Betapropiolactone (BPL) is a disinfectant, effective against bacteria, fungi and viruses. It is also used to prepare inactivated vaccines because it destroys the nucleic acid core of viruses but does not damage the capsid. For the validation of BPL when used as an inactivant, it is more important to assure the quality of inactivating agent and the validity of the inactivation process. In the present study, the inactivation kinetics of IBR virus was determined with different concentration of BPL (1:250, 1:500, 1:1000, 1:1500, 1:2000 and 1:2500) at 4 and 37 °C. The result indicated that the BPL at 4 °C was able to inactivate the IBR virus within 4, 5 and 12 h with the concentration of 1:250, 1:500 and 1:1000, respectively. BPL at 37 °C was able to inactivate virus within 30 min with the concentration of 1:250. BPL with the concentration of 1:500 and 1:1000 were able to inactivate the virus within 120 min at 37 °C. Based on the kinetic study seven formulations were prepared and a sero conversion study of IBR inactivated vaccine was carried out. Serological response in animals to different formulations did not differ significantly (P > 0.05).     

The stability of the reactive oxygen metabolites (d-ROMs) and biological antioxidant potential (BAP) tests on stored horse blood

Increasing interest in the role of oxidative stress (OS) in equine medicine has highlighted the need to develop reliable methods to quantify it. In this study we describe the effect of refrigeration (at 4 °C) on the stability of the reactive oxygen metabolites (d-ROMs) and biological antioxidant potential (BAP) tests carried out on 15 healthy horses. Blood samples, collected from the jugular vein, were immediately placed on ice and analysed using both the d-ROMs and BAP tests. Samples were also refrigerated at 4 °C and tested after 3, 7 and 24 h. The average results were similar for up to 24 h and minimal variations were found for each horse. The findings suggest that refrigeration is suitable for preserving equine blood samples for these assays and this approach will provide veterinarians with a technically simple, reliable test to measure OS under field conditions.     

In vivo efficacy of tamarind (Tamarindus indica) fruit extract on experimental fluoride exposure in rats

The study was undertaken to determine the efficacy of hydro-methanolic (1:1) extract of tamarind (Tamarindus indica L.) fruit pulp in removing body fluoride burden. Thirty rats were divided into five groups. Keeping no fluoride group as the control, rats of no treatment, low dose, middle dose and high dose groups received sodium fluoride orally at the rate of 200 mg per kg body weight daily for 14 weeks. Rats of low dose, middle dose and high dose group simultaneously received tamarind fruit pulp extract at three doses, viz. 25 (low), 50 (medium) and 100 mg (high) per kg body weight orally, respectively. Fluoride concentration in blood, urine and long bone of experimental rats was monitored to assess the efficacy of the extract. Mean serum fluoride concentration in fluoride exposed rats was 0.145 ± 0.009 and 0.783 ± 0.042 μg/ml on days 0 and 98. In comparison, fluoride concentrations in tamarind treated rats were 0.179 ± 0.021 and 0.633 ± 0.015; 0.179 ± 0.021 and 0.502 ± 0.025 and 0.176 ± 0.021 and 0.498 ± 0.030 μg/ml in low, medium and high dose groups, respectively on day 0 and day 98 of the experiment. There was a significant (p ⩽ 0.01) increase in urinary fluoride excretion from day 28 onwards. The mean fluoride concentration in long bones of treated rats was significantly lower than the values recorded in fluoride exposed rats. These findings suggest that concomitant use of tamarind fruit pulp extract can reduce fluoride concentration in blood and bone and enhanced urinary excretion, indicating the ameliorative potential of fruits of tamarind in fluoride toxicity.     

Antinociceptive efficacy and plasma concentrations of transdermal buprenorphine in dogs

To assess the antinociceptive efficacy of transdermal (TD) buprenorphine (B) in dogs, a prospective, positive-controlled experimental study was performed in 10 healthy Beagles. In an open label crossover design, the dogs initially received intravenous B (IVB, 0.02 mg kg^?1) as a positive control, followed by TDB (52.5 μg h^?1) 4 months later. Blood was collected at regular intervals for determination of the plasma concentrations of B ([B]) and its metabolite norbuprenorphine. The antinociceptive efficacy was assessed using thermal and mechanical models of nociception. The peak concentration [B] was 1.54 ng mL^?1 (±1.98) 60 h after TDB application, although three dogs had no measurable [B] after TDB. Maximum thermal threshold (TT) was 52.6 °C (±0.48) at 1 h after IVB administration and 51.63 °C (±1.01) 72 h after TDB application. The significant increase in TT indicated that effective antinociception was achieved beyond 36 h after the application of TDB, lasting until patch removal. There was hysteresis between [B] and the antinociceptive effect.     

Saliva chromogranin A in growing pigs: A study of circadian patterns during daytime and stability under different storage conditions

Salivary chromogranin A (CgA) is considered to be a biomarker of activation of the sympatho-adrenomedullary system, and has recently been proposed as a useful indicator of the acute stress response in pigs. The aim of the present study was to determinate whether salivary CgA concentrations in healthy growing pigs exhibits any circadian pattern during the daytime, and to evaluate its stability under different storage conditions. A total of 80 pigs (40 in spring and another 40 in autumn) of two different ages and genders were used. To establish the circadian pattern, saliva samples were collected at 07.00, 11.00, 15.00 and 19.00 h on two consecutive days. Pooled samples were used for the stability study and were measured on the day of sampling and periodically for up to 360 days later. Samples were stored at 4 °C, ?20 °C or ?80 °C and the effect of repeated freezing and thawing was also evaluated.No circadian pattern was detected for salivary CgA in either season and there were no significant effects of gender or age. However, mean salivary CgA concentrations were significantly higher (P < 0.0001) in the pigs sampled in autumn, compared to those sampled in the spring. Short term storage at 4 °C is recommended for up to 2 days, whereas frozen samples can be stored for 1 year at ?20 °C or ?80 °C, without substantial reduction in CgA values. In addition, samples can be frozen and thawed up to seven times without significant loss of the biomarker.     

Cryptosporidium scrofarum n. sp. (Apicomplexa: Cryptosporidiidae) in domestic pigs (Sus scrofa)

We describe the morphological, biological, and molecular characteristics of Cryptosporidium pig genotype II and propose the species name Cryptosporidium scrofarum n. sp. to reflect its prevalence in adult pigs worldwide. Oocysts of C. scrofarum are morphologically indistinguishable from C. parvum, measuring 4.81–5.96 μm (mean = 5.16) × 4.23–5.29 μm (mean = 4.83) with a length to width ratio of 1.07 ± 0.06 (n = 400). Oocysts of C. scrofarum obtained from a naturally infected pig were infectious for 8-week-old pigs but not 4-week-old pigs. The prepatent period in 8-week-old Cryptosporidium-naive pigs was 4–6 days and the patent period was longer than 30 days. The infection intensity of C. scrofarum in pigs was generally low, in the range 250–4000 oocysts per gram of feces. Infected pigs showed no clinical signs of cryptosporidiosis and no pathology was detected. Cryptosporidium scrofarum was not infectious for adult SCID mice, adult BALB/c mice, Mongolian gerbils (Meriones unguiculatus), southern multimammate mice (Mastomys coucha), yellow-necked mice (Apodemus flavicollis), or guinea pigs (Cavia porcellus). Phylogenetic analyses based on small subunit rRNA, actin, and heat shock protein 70 gene sequences revealed that C. scrofarum is genetically distinct from all known Cryptosporidium species.     

Identification and characterization of serum complement activity in the American crocodile (Crocodylus acutus)

Incubation of unsensitized sheep red blood cells with serum from the American crocodile (Crocodylus acutus) resulted in a concentration-dependent hemolysis. The hemolytic activity was heat-sensitive, and inhibited by EDTA in a concentration-dependent manner. The EDTA-inhibited SRBC hemolysis could be restored by the addition of excess Ca²⁺ or Mg²⁺, but not Ba²⁺ or Cu²⁺, revealing the specificity of this activity for these two divalent cations. The hemolytic activity of crocodile serum was titer-dependent, with 329 μL producing 50% of maximal SRBC hemolysis. The complement activity was also temperature-dependent, with decreased activity at lower temperatures (5–15 °C) and maximal activity occurred at 30–40 °C. The hemolysis occurred relatively slowly, with near zero activity after 10 min, 40% of activity observed within 15 min of exposure to SRBCs, and maximal activity at 30 min.     

Effects of hypotaurine,cysteamine and aminoacids solution on post-thaw microscopic and oxidative stress parameters of Angora goat semen

This study was conducted to determine the effects of cysteamine, hypotaurine and aminoacids solution (BME) on standard semen parameters, lipid peroxidation and antioxidant activities of Angora goat semen after the freeze–thawing process. Ejaculates collected from four Angora goats were evaluated and pooled at 37 °C. Semen samples, which were diluted with a Tris-based extender containing the antioxidants hypotaurine (5 mM) and cysteamine (5 mM), and an aminoacid solution (13%), and an extender containing no antioxidants (control), were cooled to 5 °C and frozen in 0.25-ml French straws in liquid nitrogen. Frozen straws were thawed individually at 37 °C for 20 s in a water bath for evaluation.Supplementation with cysteamine, hypotaurine and BME caused significant (P < 0.05) increases in sperm motility, and significant (P < 0.05) decreases in total abnormality rates in comparison to the control group. While all in vitro treatments did not affect the acrosomal abnormality rates, hypotaurine and BME but not cysteamine significantly (P < 0.05) increased the HOST results as compared to the control group. Supplementation with antioxidants and BME did not significantly affect MDA levels and CAT activity in comparison to the control group (P > 0.05). The antioxidants hypotaurine and cysteamine decreased SOD activity when compared to the BME group and controls (P < 0.001).     

In vitro and ex vivo inhibition of canine cyclooxygenase isoforms by robenacoxib: A comparative study

In vitro whole blood canine assays were used to quantify the inhibitory actions of the novel non-steroidal anti-inflammatory drug (NSAID) robenacoxib on the cyclooxygenase (COX) isoenzymes, COX-1 and COX-2, in comparison with other drugs of the NSAID class. COX-1 activity was determined by measuring serum thromboxane (Tx)B₂ synthesis in blood samples allowed to clot at 37 °C for 1 h. COX-2 activity was determined by measuring prostaglandin (PG)E₂ synthesis in blood samples incubated at 37 °C for 24 h in the presence of lipopolysaccharide. The rank order of selectivity for inhibition of COX-2 versus COX-1 (IC₅₀ COX-1:IC₅₀ COX-2) for veterinary drugs was highest with robenacoxib (128.8) compared to deracoxib (48.5), nimesulide (29.2), S+ carprofen (17.6), meloxicam (7.3), etodolac (6.6), R? carprofen (5.8) and ketoprofen (0.88). Selectivity expressed as the clinically relevant ratio IC₂₀ COX-1:IC₈₀ COX-2 was highest for robenacoxib (19.8) compared to deracoxib (2.3), S+ carprofen (2.5), R? carprofen (2.1), nimesulide (1.8), etodolac (0.76), meloxicam (0.46) and ketoprofen (0.21).An in vivo pharmacokinetic ex vivo pharmacodynamic study in the dog established dosage and concentration–effect relationships for single oral doses of robenacoxib over the dosage range 0.5–8.0 mg/kg. Values of C_max and AUC were linearly related to dosage over the tested range. Robenacoxib did not inhibit serum TxB₂ synthesis (COX-1) ex vivo at dosages of 0.5–4.0 mg/kg and produced only transient inhibition (at the 1 h and 2 h sampling times) at the 8 mg/kg dosage. All dosages of robenacoxib (0.5–8 mg/kg) produced marked, significant and dose related inhibition of PGE₂ synthesis (COX-2) ex vivo.The data demonstrate that in the dog robenacoxib is a highly selective inhibitor of the COX-2 isoform of COX, and significantly inhibits COX-2 and spares COX-1 in vivo when administered orally over the dosage range 0.5–4.0 mg/kg.     

Estimation of genetic parameters of racing performance in Iranian Thoroughbred horses

The aim of this study was to estimate the genetic parameters of racing performance and investigate the fixed effects that influence the performance of Thoroughbred race horses. The data included the performance of two, three and four year old horses raced in flat races and consisted of 10,800 time records and 10,912 ranking value. Two criteria were used in order to analyze the performance: racing time and a normalized ranking value. The effects of age of horse at racing and post position were significant for all traits (P < 0.05). The effect of hippodrome also was significant for racing time (P < 0.001). The genetic parameters were estimated by the restricted maximum likelihood (REML) method using DFREML program. The heritability estimates for racing time at 1000, 1400 and + 1600 m were 0.13 (± 0.009), 0.11 (± 0.007) and 0.09 (± 0.009) and for ranking value were 0.15 (± 0.010), 0.17 (± 0.008), 0.13 (± 0.009) and 0.11 (± 0.013) for entire data set, 1000, 1400 and + 1600 m respectively. Estimates of repeatability of racing time for 1000, 1400 and + 1600 m were 0.26, 0.19 and 0.17; and for ranking value were 0.28, 0.32, 0.25 and 0.20 for entire data set, 1000, 1400 and + 1600 m respectively. The genetic correlation of two criteria was 0.68–0.79 depended on distances of races.     

Pharmacokinetic,metabolism and withdrawal time of orphenadrine in camels (Camelus dromedarius) after intravenous administration

The pharmacokinetics of orphenadrine (ORPH) following a single intravenous (i.v.) dose was investigated in six camels (Camelus dormedarius). Orphenadrine was extracted from the plasma using a simple sensitive liquid–liquid extraction method and determined by gas chromatography/mass spectrometry (GC/MS). Following i.v. administration plasma concentrations of ORPH decline bi-exponentially with distribution half-life (t_1/2α) of 0.50 ± 0.07 h, elimination half-life (t_1/2β) of 3.57 ± 0.55 h, area under the time concentration curve (AUC) of 1.03 ± 0.10 g/h l⁻¹. The volume of distribution at steady state (Vd_ss) 1.92 ± 0.22 l kg⁻¹, volume of the central compartment of the two compartment pharmacokinetic model (V_c) 0.87 ± 0.09 l kg⁻¹, and total body clearance (Cl_T) of 0.60 ± 0.09 l/h kg⁻¹. Three orphenadrine metabolites were identified in urine samples of camels. The first metabolite N-desmethyl-orphenadrine resulted from N-dealkylation of ORPH with molecular ion m/z 255. The second N,N-didesmethyl-orphenadrine, resulted from N-didesmethylation with molecular ion m/z 241. The third metabolite, hydroxyl-orphenadrine, resulted from the hydroxylation of ORPH with molecular ion m/z 285. ORPH and its metabolites in camel were extensively eliminated in conjugated form. ORPH remains detectable in camel urine for three days after i.v. administration of a single dose of 350 mg orphenadrine aspartate.     

Airway hyperresponsiveness to adenosine 5’-monophosphate in feline chronic inflammatory lower airway disease

Airway hyperresponsiveness is a key feature of human asthma and chronic bronchitis and response to the indirectly acting agonist adenosine 5′-monophosphate (AMP) is thought to reflect underlying airway inflammation. To examine whether airway responsiveness testing (ART) with AMP may be used to differentiate healthy cats from those with asthma (FA) and chronic bronchitis (CB), 24 cats (9 FA, 6 CB, 9 controls) underwent ART with AMP at concentrations of 0.1, 1, 10, 100 and 500 mg/mL using barometric whole body plethysmography.The defined endpoint of ART, an increase in enhanced pause (Penh) exceeding 300% of the post-saline value (baseline), was reached in 9/15 patients (7 FA, 2 CB), but in none of the controls. Mean Penh (±SD) at baseline (BL) was 0.49 ± 0.16 for cases, and 0.54 ± 0.16 for controls, and was significantly increased after AMP challenge in clinical cases (2.62 ± 2.20), but not in controls (0.63 ± 0.30, P < 0.05). After separating responder (R) and non-responder (NR) cases, a more pronounced difference after challenge was found (R: 3.96 ± 1.84, NR: 0.6 ± 0.21, P < 0.001). The provocative concentration of the agonist that increased Penh to 300% of BL (PC Penh 300) in R cases was 52.98 ± 48.04 mg/mL AMP. Age had no influence on the responder status or PC Penh 300. It was concluded that AMP challenge may offer a new method for the identification of cats with lower inflammatory airway disease, and possibly for monitoring disease progression or response to therapy.     

Novel and simple approach using synthesized nickel nanoparticles to control blood-sucking parasites

The present study was on assessment of the anti-parasitic activities of nickel nanoparticles (Ni NPs) against the larvae of cattle ticks Rhipicephalus (Boophilus) microplus and Hyalomma anatolicum (a.) anatolicum (Acari: Ixodidae), fourth instar larvae of Anopheles subpictus, Culex quinquefasciatus and Culex gelidus (Diptera: Culicidae). The metallic Ni NPs were synthesized by polyol process from Ni-hydrazine as precursor and Tween 80 as both the medium and the stabilizing reagent. The synthesized Ni NPs were characterized by Fourier transform infrared (FTIR) spectroscopy analysis which indicated the presence of Ni NPs. Synthesized Ni NPs showed the X-ray diffraction (XRD) peaks at 42.76°, 53.40°, and 76.44°, identified as 1 1 1, 2 2 0, and 2 0 0 reflections, respectively. Scanning electron microscopy (SEM) analysis of the synthesized Ni NPs clearly showed that the Ni NPs were spherical in shape with an average size of 150 nm. The Ni NPs showed maximum activity against the larvae of R. (B.) microplus, H. a. anatolicum, A. subpictus, C. quinquefasciatus and C. gelidus with LC₅₀ values of 10.17, 10.81, 4.93, 5.56 and 4.94 mg/L; r² values of 0.990, 0.993, 0.992, 0.950 and 0.988 and the efficacy of Ni-hydrazine complexes showed the LC₅₀ values of 20.35, 22.72, 8.29, 9.69 and 7.83 mg/L; r² values of 0.988, 0.986, 0.989, 0.944 and 0.978, respectively. The findings revealed that synthesized Ni NPs possess excellent larvicidal parasitic activity. To the best of our knowledge, this is the first report on larvicidal activity of blood feeding parasites using synthesized Ni NPs.     

Nutritional management of inherited copper-associated hepatitis in the Labrador retriever

Canine hereditary copper-associated hepatitis is characterized by gradual hepatic copper accumulation eventually leading to liver cirrhosis. Therapy is aimed at creating a negative copper balance with metal chelators, of which d-penicillamine is the most commonly used. d-penicillamine often causes gastro-intestinal side effects and life-long continuous therapy may lead to a deficiency of copper and zinc. The aim of the current study was to investigate the effect of a low-copper, high-zinc diet as an alternative to continuous d-penicillamine treatment for the long-term management of canine copper-associated hepatitis.Sixteen affected Labrador retrievers were followed for a median time period of 19.1 months (range, 5.9–39 months) after being effectively treated with d-penicillamine. The dogs were maintained on a diet containing 1.3 ± 0.3 mg copper/1000 kcal and 64.3 ± 5.9 mg zinc/1000 kcal. Liver biopsies were taken every 6 months for histological evaluation and copper determination. Plasma alanine aminotransferase (ALT) and alkaline phosphatase, as well as serum albumin were determined.Dietary treatment alone was sufficient to maintain hepatic copper concentration below 800 mg/kg dry weight liver in 12 dogs during the study period. Four dogs needed re-treatment with d-penicillamine. ALT activity and albumin concentration were not associated with hepatic copper concentration, but showed a significant association with the stage and grade of hepatitis respectively. In conclusion, a low-copper, high-zinc diet can be a valuable alternative to continuous d-penicillamine administration for long-term management of dogs with copper-associated hepatitis. The copper re-accumulation rate of an individual dog should be considered in the design of a long-term management protocol and in determining re-biopsy intervals.     

Effects of Pre-Weaning Vitamin E,Selenium, and Copper Supplementation on the Performance,Acute Phase Protein Concentration,and Immune Function of Stressed Beef Calves

Two experiments were conducted to determine the effects of pre-weaning vitamin E, Se, and Cu supplementation on performance and immune response in stressed calves. In Exp. 1, 71 Hereford x Angus calves were individually creep fed: 1) control supplement (CON), 2) control plus 500 IU vitamin E + 0.3 mg Se/kg DM (E), 3) control plus 10 mg Cu/kg DM (CU), or 4) a combination of E and CU treatments (ECU). In Exp. 2, 80 Hereford (Angus calves were individually creep fed: 1) control supplement (CON), 2) control plus 0.3 mg Se/kg DM (SE), 3) control plus 500 IU vitamin E + 0.3 mg Se/kg DM (LOWE), 4) control plus 1000 IU vitamin E + 0.3 mg Se/kg DM (MEDE), or 5) control plus 1500 IU vitamin E + 0.3 mg Se/kg DM (HIE). Treatments continued for 49 (Exp. 1) or 53 d (Exp. 2) prior to weaning. At weaning all calves were transported to feedlot facilities. In Exp. 1, vitamin E tended (P<0.09) to improve post-weaning ADG and reduce (P<0.06) plasma haptoglobin (Hp), but had no effect on plasma α-tocopherol. Dietary Cu tended to increase (P<0.01) liver Cu stores, and antibody titers to bovine viral diarrhea (BVD) were greater (P<0.04) at weaning in CU and E calves. In Exp. 2, vitamin E tended to increase serum α-tocopherol (P<0.06) and cortisol (P<0.08). Vitamin E and Se supplementation may improve post-weaning performance and decrease plasma Hp concentrations in stressed calves.     

Estimation of direct and maternal (co)variance components for pre-weaning growth traits in Muzaffarnagari sheep

Genetic parameters and (co)variance components were estimated for weight at birth and at 15, 30, 45, 60 and 75 days of age for a flock of Muzaffarnagari sheep maintained at the Central Institute for Research on Goats, Makhdoom, Mathura over a period of 27 years (1976–2002). Records on 5201 lambs descended from 1568 ewes and 170 rams were included in the analysis. Analyses were carried out by REML fitting an animal model and ignoring or including maternal genetic or permanent environmental effects. Six different animal models were fitted for all traits, and the best model was chosen after testing improvements in log-likelihood values. Direct heritability estimates were inflated substantially for all traits when maternal effects were ignored. Direct heritability estimates were 0.08 ± 0.02 for birth weight and 0.02 ± 0.02, 0.02 ± 0.02, 0.27 ± 0.08, 0.09 ± 0.04, and 0.29 ± 0.08 for weights at 15, 30, 45, 60, and 75 days, respectively. Maternal genetic effects contributed only 4 to 8% of the total phenotypic variance from birth to 30 days of age, and this effect diminished further with increasing age. Maternal heritability was low for pre-weaning growth traits and should have only a small effect on selection response. Estimates of the fraction of variance due to maternal permanent environmental effects were 0.09 ± 0.02, 0.15 ± 0.04, 0.12 ± 0.03, 0.11 ± 0.04, 0.14 ± 0.02, and 0.08 ± 0.04 for body weights at birth and at 15, 30, 45, 60, and 75 days, respectively. These results indicate that selecting for improved maternal and/or direct effects in Muzaffarnagari sheep would generate only slow genetic progress in early growth traits.     

Isolation of active compounds from methanol extracts of Toddalia asiatica against Ichthyophthirius multifiliis in goldfish (Carassius auratus)

The parasitic ciliate Ichthyophthirius multifiliis infests all species of freshwater fish and can cause severe economic losses in fish breeding. The present study aims to evaluate the antiparasitic activity of the active components from Toddalia asiatica against I. multifiliis. Bioassay-guided fractionation and isolation of compounds with antiparasitic activity were performed on the methanol extract of T. asiatica yielding two bioactive compounds: chelerythrine and chloroxylonine identified by comparing spectral data (NMR and ESI-MS) with literature values. Results from in vitro antiparasitic assays revealed that chelerythrine and chloroxylonine could be 100% effective against I. multifiliis at the concentration of 1.2 mg L^?1 and 3.5 mg L^?1, with the median effective concentration (EC₅₀) values of 0.55 mg L^?1 and 1.90 mg L^?1 respectively. In vivo experiments demonstrated that fish treated with chelerythrine and chloroxylonine at the concentrations of 1.8 and 8.0 mg L^?1 carried significantly fewer parasites than the control (P < 0.05). The acute toxicity (LC₅₀) of chelerythrine for goldfish was 3.3 mg L^?1.