In vitro antiviral activity of chestnut and quebracho woods extracts against avian reovirus and metapneumovirus

Field evidences have suggested that a natural extract, containing tannins, could be effective against poultry enteric viral infections. Moreover previous studies have shown that vegetable tannins can have antiviral activity against human viruses. Based on this knowledge three different Chestnut (Castanea spp.) wood extracts and one Quebracho (Schinopsis spp.) wood extract, all containing tannins and currently used in the animal feed industry, were tested for in vitro antiviral activity against avian reovirus (ARV) and avian metapneumovirus (AMPV). The MTT assay was used to evaluate the 50% cytotoxic compounds concentration (CC₅₀) on Vero cells. The antiviral properties were tested before and after the adsorption of the viruses to Vero cells. Antiviral activities were expressed as IC₅₀ (concentration required to inhibit 50% of viral cytopathic effect). CC₅₀s of tested compounds were >200 μg/ml. All compounds had an extracellular antiviral effect against both ARV and AMPV with IC₅₀ values ranging from 25 to 66 μg/ml. Quebracho extract had also evident intracellular anti-ARV activity (IC₅₀ 24 μg/ml). These preliminary results suggest that the examined vegetable extracts might be good candidates in the control of some avian virus infections. Nevertheless further in vivo experiments are required to confirm these findings.     

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.     

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.     

Acaricidal properties of two extracts from Guiera senegalensis J.F. Gmel. (Combrataceae) against Hyalomma anatolicum (Acari: Ixodidae)

Laboratory test were carried out on eggs, larvae, nymphs and adults of Hyalomma anatolicum to determine the acaricidal activities of petroleum ether (PE) and crude ethanolic extracts (EE) from the leaves of Guiera senegalensis J.F. Gmel. (Combrataceae) using immersion method.Stock solutions, of 300 mg/ml (30%) of the each two extract, were prepared. Six two-fold serial dilutions each with three replicates were used. Both extracts, at the highest concentration 150 mg/ml (15%), induced 100% failure of hatching of the treated eggs. The concentrations of PE and EE that induced 50% inhibition of the hatchability (IC₅₀) were 1.71 and 0.508%, respectively. In the larval immersion test (LIT), EE at 15% concentration caused complete mortality while the same concentration of PE resulted in 96% mortality. The mortalities increased with concentrations. There was a correlation between the mortalities and increased concentrations, the values of the linear correlation coefficient (r) for PE and EE were 0.93 and 0.79, and The LC₅₀ and LC₉₉ were 2.08 and 14.09, and 0.787 and 11.054, respectively. At the concentrations of 3.75%, 7.5% and 15%, PE inhibited the molting of the nymphs by 40, 55 and 65%, respectively, while EE induced 46.49, 64.3 and 71.4% inhibition, respectively. The effectiveness of the treatment against unfed adult females was assessed by measuring the feeding performance and egg production using adult immersion test (AIT). Although, there was no mortality in unfed adults, PE and EE inhibited feeding and egg-laying of the survived females by 35–100% and 6.16–100%, respectively. Our results indicated that G. senegalensis is a promising biocontrol candidate as an acaricidal agent against H. anatolicum.     

Pharmacokinetics and ex vivo pharmacodynamics of cefquinome in porcine serum and tissue cage fluids

A tissue cage (TC) model was used to evaluate the pharmacokinetics and ex vivo pharmacodynamics of cefquinome after intravenous (IV) and intramuscular (IM) administration to piglets at 2 mg/kg bodyweight. The mean values of area under the concentration–time curve (AUC) were 21.28 (IV) and 21.37 (IM) μg h/mL for serum, and 17.40 (IV) and 16.57 (IM) μg h/mL for TC fluid (TCF), respectively. Values of maximum concentration (C_max) were 6.15 μg/mL (serum) and 1.15 μg/mL (TCF) after IM administration. The elimination half-lives (t_1/2β) in TCF (10.63 h IV and 11.81 h IM) were significantly higher than those in serum (2.33 h IV and 2.30 h IM) (P < 0.05). The values of AUC_TCF/AUC_serum (%) after IV and IM administration were 82.4% and 80.7%, respectively.The ex vivo time-kill curves were established for serum and TCF samples using Escherichia coli ATCC 25922. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration values of cefquinome against E. coli were 0.030 and 0.060 μg/mL in Mueller–Hinton broth, and 0.032 and 0.064 μg/mL in both serum and TCF, respectively. The ex vivo growth inhibition data of TCF after IM administration were fitted to the sigmoid E_max model; AUC_24h/MIC was 35.01 h for bactericidal activity and 44.28 h for virtual eradication, respectively. The findings from this study suggest that cefquinome may be therapeutically effective in diseases of pigs caused by E. coli when used at a dose rate of 1.33 mg/kg administered every 24 h for organisms with MIC₉₀ ⩽ 0.50 μg/mL.     

Evaluation of in vitro and in vivo inhibitory effects of fusidic acid on Babesia and Theileria parasites

Fusidic acid known to has antibacterial, antifungal, and antimalarial activities. Fusidic acid blocks translation elongation factor G gene in Plasmodium falciparum. In the present study, the inhibitory effects of fusidic acid on the in vitro growth of bovine and equine Babesia parasites were evaluated. The inhibitory effect of fusidic acid on the in vivo growth of Babesia microti was also assessed. The in vitro growth of four Babesia species that were tested was significantly inhibited (P < 0.05) by micromolar concentrations of fusidic acid (IC₅₀ values = 144.8, 17.3, 33.3, and 56.25 μM for Babesia bovis, Babesia bigemina, Babesia caballi, and Theileria equi, respectively). Combinations of fusidic acid with diminazene aceturate synergistically potentiated its inhibitory effects in vitro on B. bovis and B. caballi. In B. microti-infected mice, fusidic acid caused significant (P < 0.05) inhibition of the growth of B. microti at the dose of 500 mg/kg BW relative to control group. These results indicate that fusidic acid might be incorporated in treatment of babesiosis.     

In-vitro antiviral efficacy of ribavirin and interferon-alpha against canine distemper virus

Canine distemper is a highly contagious disease with high incidence and lethality in the canine population. The objective of this study was to evaluate the efficacy of antiviral action with ribavirin (RBV), interferon-alpha (IFNα), and combinations of RBV and IFNα against canine distemper virus (CDV). Vero cells inoculated with CDV were treated with RBV, IFNα, and combinations of these drugs. The efficacy to inhibit viral replication was evaluated by adding the compounds at different times to determine which step of the viral replicative process was affected. Both drugs were effective against CDV in vitro. The IFNα was the most active compound, with an average IC₅₀ (50% inhibitory concentration) value lower than the IC₅₀ of the RBV. Ribavirin (RBV) was more selective than IFNα, however, and neither drug showed extracellular antiviral activity. The combination of RBV and IFNα exhibited antiviral activity for the intra- and extracellular stages of the replicative cycle of CDV, although the intracellular viral inhibition was higher. Both RBV and IFNα showed high antiviral efficacy against CDV, and furthermore, RBV + IFNα combinations have shown greater interference range in viral infectivity. These compounds could potentially be used to treat clinical disease associated with CDV infection.     

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.     

In vitro assessment of the antiviral potential of trans-cinnamic acid,quercetin and morin against equid herpesvirus 1

The antiviral activity of quercetin, morin and trans-cinnamic acid was evaluated in vitro against equid herpesvirus 1 (EHV-1) by determining the virucidal activity and using the time of addition assay to test inhibition of the viral replication cycle. The cytotoxicity of each substance was assessed using MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide]. Quercetin showed virucidal action and inhibition of the viral replication cycle at 0 and 1 h. Morin showed potential virucidal and viral replication cycle inhibition at 0 h. Trans-cinnamic acid did not show virucidal activity but inhibited the viral replication cycle at −1 and 0 h. This study demonstrates the potential of these compounds as future antiviral candidates in relation to viruses of importance in veterinary medicine.     

Potency and selectivity of carprofen enantiomers for inhibition of bovine cyclooxygenase in whole blood assays

Whole blood in vitro assays were used to determine the potency and selectivity of carprofen enantiomers for inhibition of the isoforms of cyclooxygenase (COX), COX-1 and COX-2, in the calf. S(+)-carprofen possessed preferential activity for COX-2 inhibition but, because the slopes of inhibition curves differed, the COX-1:COX-2 inhibition ratio decreased from 9.04:1 for inhibitory concentration (IC)₁₀ to 1.84:1 for IC₉₅. R(−) carprofen inhibited COX-2 preferentially only for low inhibition of the COX isoforms (IC₁₀ COX-1:COX-2 = 6.63:1), whereas inhibition was preferential for COX-1 for a high level of inhibition (IC₉₅ COX-1:COX-2 = 0.20:1). S(+) carprofen was the more potent inhibitor of COX isoforms; potency ratios S(+):R(−) carprofen were 11.6:1 for IC₁₀ and 218:1 for IC₉₀. Based on serum concentrations of carprofen enantiomers obtained after administration of a therapeutic dose of 1.4 mg/kg to calves subcutaneously, S(+)-carprofen concentrations exceeded the in vitro IC₈₀ COX-2 value for 32 h and the IC₂₀ for COX-1 for 33 h. The findings are discussed in relation to efficacy and safety of carprofen in calves.     

The use of eugenol against Aeromonas hydrophila and its effect on hematological and immunological parameters in silver catfish (Rhamdia quelen)

The aim of this study was to evaluate the activity of eugenol against the fish pathogen Aeromonas hydrophila and eugenol's effect on hematological and natural immune parameters in silver catfish (Rhamdia quelen). In vitro, eugenol showed weak activity against A. hydrophila, but in vivo, at a subinhibitory concentration (10 mg L^?1), it promoted survival in infected silver catfish. Eugenol (50 μg mL^?1) reduced the hemolytic activity of A. hydrophila supernatant in vitro in fish erythrocytes. Subjecting catfish to eugenol baths (5 and 10 mg L^?1) for five days did not alter the hematological and immunological parameters studied in this work. Based on these results, eugenol can be used to treat or prevent bacterial diseases in fish.     

In vitro inhibition of Eimeria tenella invasion of epithelial cells by phytochemicals

Resistance to coccidiostats and possible future restrictions on their use raise the need for alternative methods of reducing coccidiosis in poultry. The aim of this study was to evaluate the effect of selected phytochemicals on Eimeria tenella sporozoite invasion in vitro. Four phytochemicals were selected on the basis that they reduce the virulence of Eimeria spp. and/or provide immune modulatory benefits to host cells: betaine, carvacrol, curcumin and Echinacea purpurea extract (EP).Madin–Darby bovine kidney (MDBK) cells were covered by medium containing phytochemicals at the highest concentration which was non-toxic to the cells. Salinomycin 50 μg/ml was positive control; negative control was medium only. E. tenella (Houghton strain) sporozoites were added to wells and after incubation for 2, 4 or 20 h at 37 °C, cells were fixed and stained with hematoxylin–eosin. Ten evenly spaced fields per well were photographed and the percentage of cells invaded by sporozoites was calculated and normalized to the control.At 2 h, carvacrol, curcumin and EP showed a significantly lower percentage of sporozoite invasion than the untreated control; in contrast, betaine treatment represented a significantly higher invasion percentage. Combining carvacrol with EP inhibited E. tenella invasion more effectively than applying the compounds individually, but the further addition of curcumin did not reduce invasion further.In conclusion, this study shows that invasion of MDBK epithelial cells by E. tenella sporozoites is inhibited in the presence of carvacrol, curcumin, or EP and enhanced by betaine. There may be potential for developing these phytochemicals as anti-coccidial feed or water additives for poultry.     

Pharmacokinetics of erythromycin after intravenous,intramuscular and oral administration to cats

The aim of this study was to characterise the pharmacokinetic properties of different formulations of erythromycin in cats. Erythromycin was administered as lactobionate (4 mg/kg intravenously (IV)), base (10 mg/kg, intramuscularly (IM)) and ethylsuccinate tablets or suspension (15 mg/kg orally (PO)). After IV administration, the major pharmacokinetic parameters were (mean ± SD): area under the curve (AUC)_(0–∞) 2.61 ± 1.52 μg h/mL; volume of distribution (V_z) 2.34 ± 1.76 L/kg; total body clearance (Cl_t) 2.10 ± 1.37 L/h kg; elimination half-life (t_½λ) 0.75 ± 0.09 h and mean residence time (MRT) 0.88 ± 0.13 h. After IM administration, the principal pharmacokinetic parameters were (mean ± DS): peak concentration (C_max), 3.54 ± 2.16 μg/mL; time of peak (T_max), 1.22 ± 0.67 h; t_½λ, 1.94 ± 0.21 h and MRT, 3.50 ± 0.82 h. The administration of erythromycin ethylsuccinate (tablets and suspension) did not result in measurable serum concentrations. After IM and IV administrations, erythromycin serum concentrations were above minimum inhibitory concentration (MIC)₉₀ = 0.5 μg/mL for 7 and 1.5 h, respectively. However, these results should be interpreted cautiously since tissue erythromycin concentrations have not been measured and can reach much higher concentrations than in blood, which may be associated with enhanced clinical efficacy.     

Vaccination with a genotype 1 modified live vaccine against porcine reproductive and respiratory syndrome virus significantly reduces viremia,viral shedding and transmission of the virus in a quasi-natural experimental model

The present study assessed the efficacy of vaccination against genotype 1 porcine reproductive and respiratory syndrome virus (PRRSV) in terms of reduction of the transmission. Ninety-eight 3-week-old piglets were divided in two groups: V (n = 40) and NV (n = 58) that were housed separately. V animals were vaccinated with a commercial genotype 1 PRRSV vaccine while NV were kept as controls. On day 35 post-vaccination, 14 NV pigs were separated and inoculated intranasally with 2 ml of a heterologous genotype 1 PRRSV isolate (“seeder” pigs, SP). The other V and NV animals were distributed in groups of 5 pigs each. Two days later, one SP was introduced into each pen to expose V and NV to PRRSV. Sentinel pigs were allocated in adjacent pens. Follow-up was of 21 days. All NV (30/30) became viremic after contact with SP while only 53% of V pigs were detected so (21/40, p < 0.05). Vaccination shortened viremia (12.2 ± 4 versus 3.7 ± 3.4 days in NV and V pigs, respectively, p < 0.01). The 50% survival time for becoming infected (Kaplan–Meier) for V was 21 days (CI_95% = 14.1–27.9) compared to 7 days (CI_95% = 5.2–8.7) for NV animals (p < 0.01). These differences were reflected in the R value as well: 2.78 (CI_95% = 2.13–3.43) for NV and 0.53 (CI_95% = 0.19–0.76) for V pigs (p < 0.05). All sentinel pigs (10/10) in pens adjacent to NV + SP pens got infected compared to 1/4 sentinel pigs allocated contiguous to a V + SP pen. These data show that vaccination of piglets significantly decrease parameters related to PRRSV transmission.     

Efficacy of condensed tannins against larval Hymenolepis diminuta (Cestoda) in vitro and in the intermediate host Tenebrio molitor (Coleoptera) in vivo

Natural anti-parasitic compounds in plants such as condensed tannins (CT) have anthelmintic properties against a range of gastrointestinal nematodes, but for other helminths such effects are unexplored. The aim of this study was to assess the effects of CT from three different plant extracts in a model system employing the rat tapeworm, Hymenolepis diminuta, in its intermediate host, Tenebrio molitor. An in vitro study examined infectivity of H. diminuta cysticercoids (excystation success) isolated from infected beetles exposed to different concentrations of CT extracts from pine bark (PB) (Pinus sps), hazelnut pericarp (HN) (Corylus avellana) or white clover flowers (WC) (Trifolium repens), in comparison with the anthelmintic drug praziquantel (positive control). In the in vitro study, praziquantel and CT from all three plant extracts had dose-dependent inhibitory effects on cysticercoid excystation. The HN extract was most effective at inhibiting excystation, followed by PB and WC. An in vivo study was carried out on infected beetles (measured as cysticercoid establishment) fed different doses of PB, HN and praziquantel. There was a highly significant inhibitory effect of HN on cysticercoid development (p = 0.0002). Overall, CT showed a promising anti-cestodal effect against the metacestode stage of H. diminuta.     

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.     

Comparative pharmacokinetics of an injectable cephalexin suspension in beef cattle

This study describes and compares the pharmacokinetics of a single 7.5 mg/kg dose of cephalexin monohydrate oil-based 20% suspension after its administrations to six cows by the intramuscular (i.m.) and subcutaneous (s.c.) routes, and to five calves by the i.m. route. Significantly (P < 0.05) higher peak plasma concentrations (5.6 ± 0.79 μg/ml versus 3.93 ± 1.24 μg/ml) and lower half-life (1.81 ± 0.56 h versus 4.21 ± 0.82 h) and mean residence time (4.12 ± 1.07 h versus 6.63 ± 0.85 h) were obtained after i.m. administration when compared to the s.c. administration to cows. No differences were found between pharmacokinetic parameters calculated for cows and calves. Cephalexin plasma concentrations remained above 0.5–0.75 μg/ml for 11–14 h and 8–9 h after the s.c. and i.m. administrations, respectively. Thus, route of administration may be an important issue to be considered when calculating dosage schedules for successful treatments and safe withdrawal times for veterinary medicines.