Stability study of veterinary drugs in standard solutions for LC-MS/MS screening in food.

A study on stability of veterinary drugs in standard solutions stored at -80°C and at -20°C was conducted over 1 year. Data were acquired on 152 individual stock standard solutions and also on 15 family mixes and 2 working standard solutions. All solutions were prepared, stored and compared 1 year later against freshly prepared ones by LC-MS/MS. A statistical analysis was performed to set the acceptability criteria, taking into account the variability of standard preparations. In individual stock standard solutions stored at -80°C (12 months) and -20°C (9 months), stability was demonstrated for 141 and 140 out of 152 compounds, i.e. for 92% and 93% of compounds, respectively. Drugs were even more stable when solubilised in either diluted family mixes or working standard solutions, with more than 99% and 94% of compounds found unaltered when stored at -80°C and at -20°C, respectively. In mixes, beta-lactams from the cephalosporin (cefadroxil and cephalexin) and penicillin (amoxicillin and ampicillin) families were found to be the least stable compounds when stored at -20°C (6 months), necessitating storage at -80°C to achieve a 1-year shelf life. The study also evidenced solubility issues for two sulfonamides (sulfadiazine and sulfamerazine) in methanol-based solutions. An independent stability study conducted by a second laboratory confirmed the 1-year stability of 3 family mixes-quinolones, sulfonamides and tetracyclines.

Assessment of a Flavone-Polysaccharide Based Prescription for Treating Duck Virus Hepatitis.

Because polysaccharide and flavone ingredients display good antiviral activity, we developed a flavone/polysaccharide-containing prescription that would be effective against duck viral hepatitis (DVH) and investigated its hepatoprotective effects. Flavones were derived from Hypericum japonicum (HJF) (entire herb of Hypericum japonicum Thunb) and Salvia plebeia (SPF) (entire herb of Salvia plebeia R. Br.), and polysaccharides were derived from Radix Rehmanniae Recens (RRRP) (dried root of Rehmannia glutinosa Libosch). This prescription combination was based on the theory of syndrome differentiation and treatment in traditional Chinese veterinary medicine. In vitro and in vivo experiments were conducted using the three single ingredients compared to the combined HRS prescription to determine their anti-duck hepatitis A viral (anti-DHAV) activity. The results showed that all experimental conditions displayed anti-DHAV activity, but the HRS prescription presented the best effect. To further investigate the hepatoprotective effect of the HRS prescription on DHAV-induced hepatic injury, we tested the mortality rate, the hepatic pathological severity score, plasma biochemical indexes of hepatic function, blood DHAV gene expression levels and peroxidation damage evaluation indexes and then analyzed correlations among these indexes. The results demonstrated that the HRS prescription significantly decreased the mortality rate, reduced the severity of hepatic injury, decreased the hepatic pathological severity score, depressed blood DHAV gene expression levels, and returned the indexes of hepatic function and peroxidation almost to a normal level. These results indicate that the HRS prescription confers an outstanding hepatoprotective effect, and we expect that it will be developed into a new candidate anti-DHAV drug.

Immobilization of tetracyclines in manure and manure-amended soils using aluminum-based drinking water treatment residuals.

Veterinary antibiotics (VAs) are emerging contaminants of concern in the environment, mainly due to the potential for development of antibiotic-resistant bacteria and effect on microbiota that could interfere with crucial ecosystem functions such as nutrient cycling and decomposition. High levels of VAs such as tetracyclines (TCs) have been reported in agricultural soils amended with manure, which also has the potential to cause surface and groundwater contamination. Several recent studies have focused on developing methods to immobilize VAs such as composting with straw, hardwood chips, commercial biochar, aeration, mixing, heat treatment, etc. The major shortcomings of these methods include high cost and limited effectiveness. In the current study, we assessed the effectiveness of aluminum-based drinking water treatment residuals (Al-WTR) as a "green" sorbent to immobilize TCs in manure and manure-applied soils with varying physicochemical properties by laboratory incubation study. Results show that Al-WTR is very effective in immobilizing tetracycline (TTC) and oxytetracycline (OTC). The presence of phosphate resulted in significant (p < 0.01) decrease in TTC/OTC sorption by Al-WTR, but the presence of sulfate did not. attenuated total reflection (ATR)-FTIR spectroscopy indicate that TTC and OTC likely forming surface complexes via inner-sphere-type bonds in soils, manure, and manure-applied soils amended with Al-WTR.

Characterisation of commercial Perna canaliculus samples and development of extemporaneous oral veterinary paste formulations containing Perna.

Perna canaliculus is a nutritional supplement recently studied and highly recommended for its anti-inflammatory effects in both animals and humans. In this study, the physicochemical properties, the microbiological quality, the total lipid content and fatty acids composition of three commercial samples of Perna powder were determined. Subsequently, three simple formulations of extemporaneous oral pastes containing Perna were prepared and designed for veterinary use. Their microbiological stability was assessed after 1-month storage at either room temperature or 35 °C. The results demonstrated that commercial Perna samples lack homogeneity, in regard to some technological properties and fatty acid composition; therefore, a preliminary characterisation of commercial Perna samples is recommended to assure the quality of formulations containing this nutritional supplement. Oral paste formulations are easy and simple to prepare and show good physical and microbiological stability, suggesting their large-scale production.

Development of shampoo, soap and ointment formulated by green synthesised silver nanoparticles functionalised with antimicrobial plants oils in veterinary dermatology: treatment and prevention strategies.

Many scientists have focused their research on the role of nanotechnology for the control of human pathogens, but there are also many topical pathogens present in animals, which infect animals and transfer to humans. Topical therapy is extremely important for the management of dermatological condition in animals. Therefore, the present study aims to evaluate the efficacy of biogenic silver nanoparticles (AgNPs) in combination with herbal oils against animal skin infections which may be responsible for causing infections in human beings. Here, the authors synthesised and characterised the AgNPs from Azadirachta indica. The oils were extracted from medicinal plants including Cymbopogon citratus, Cymbopogon martini, Eucalyptus globules, A. indica and Ocimum sanctum and the antifungal and antibacterial activity of plant oils along with AgNPs were evaluated. An excision wound model was used for the study of wound healing activity in rabbits. AgNPs functionalised oil has demonstrated remarkable antimicrobial activity against pathogens present on the skin of animals. The nano-functionalised antimicrobial oils were used in the formulation of shampoo, soap and ointment for veterinary dermatology. Antimicrobial products of plant origin with AgNPs are valuable, safe and have a specific role in controlling diseases. The authors believe that this approach will be a good alternative therapy to solve the continuous antibiotic resistance developed by many bacterial pathogens and will be utilised in various animal contacting areas in medicine.

Nutrient and metal analyses of Chinese herbal products marketed for veterinary use.

Many Chinese herbs and herbal mixtures are fed to domestic animals for their reputed medicinal properties. These herbs could contribute to the intake of essential nutrients and toxic metals, but their composition is mostly unknown. The purpose of this study was to measure major nutrient (crude protein, crude fat, carbohydrate, fibre) and mineral (Ca, P, Mg, K, Na, Fe, Zn, Cu, Mn, Mo, S, Al, Cd, Ni, Pb) concentrations in samples of fourteen combination formulas labelled for veterinary use and commonly administered to horses and dogs. Three single herbs, Bupleurum chinense, Curcuma zedoaria and Astragalus membranaceus, each obtained from several sources, and Yunnan Baiyao, a proprietary hemostatic mixture, were also analysed. Proximate analyses and some mineral concentrations differed (p < 0.05) among single herbs, and high concentrations of several minerals were detected in some herbal combinations. Those containing the highest concentrations [g/kg dry matter (DM)] of calcium (92.4), iron (2.6) and manganese (0.28) could provide >38%, 142% and 96%, respectively, of recommended allowances in adult dogs, and >13%, 122% and 2%, respectively, of maintenance requirements in horses, at the maximum labelled dose assuming complete availability. Concentrations of cadmium, nickel and lead were below published oral tolerance levels. Aluminium concentrations (median 380, maximum 920 mg/kg DM) were higher than has been previously reported in Chinese herbs. These nutrient analyses suggest that herbal combinations marketed to veterinarians, when fed at the maximal labelled dose, are unlikely to produce clinically relevant changes in the dietary intake of essential nutrients. However, small amounts of non-essential contaminant minerals are present in some formulas, and further research is necessary to understand the significance of this finding.

Documentation and determination of consensus about phytotherapeutic veterinary practices among the Tharu tribal community of Uttar Pradesh, India.

PURPOSE: The aim of the present investigation was to document the phytotherapeutic knowledge and veterinary healthcare management practices among the Tharu tribal community of Uttar Pradesh, India and to determine the consensus of such practices, in order to evaluate the potential for new veterinary drugs of herbal origin. METHODS: This study was conducted in 2000-2004 using semistructured, open-ended questionnaires, informal interviews and group discussions with farmers engaged in animal husbandry. RESULTS: In the present study, 59 phytotherapeutic practices using 48 plant species were documented for management of 18 types of healthcare problems of domesticated animals. Crude drug formulations keep the animal healthy, increase lactation, and reduce estrus interval and puberty period to make them economically more important. There was great agreement among informants regarding phytotherapeutic uses of medicinal plants with factor of informants' consensus (F(IC)) value ranging from 0.84 to 1, with an average value of 0.94. CONCLUSION: Study reveals that there is great agreement among informants for the usages of Azadirachta indica A Juss, Bombax ceiba L, Bambusa arundinacea (Retz) Willd, Corianderum sativum L, Cuscuta reflexa Roxb, Datura metal L, Nelumbo nucifera Gaertn, and Parthenium hysterophorus L. These species may be used for the development of new, cheep, effective, and eco-friendly herbal formulations for veterinary healthcare management. Further investigation of these herbal formulations for veterinary healthcare management will require safety and efficacy testing. There is an urgent need to formulate suitable conservation strategies for wildly growing phytotherapeutics to overcome their depletion from natural resources and to make these practices more eco-friendly.

Caenorhabditis elegans as a model to screen plant extracts and compounds as natural anthelmintics for veterinary use.

The most challenging obstacles to testing products for their anthelmintic activity are: (1) establishing a suitable nematode in vitro assay that can evaluate potential product use against a parasitic nematode of interest and (2) preparation of extracts that can be redissolved in solvents that are miscible in the test medium and are at concentrations well tolerated by the nematode system used for screening. The use of parasitic nematodes as a screening system is hindered by the difficulty of keeping them alive for long periods outside their host and by the need to keep infected animals as sources of eggs or adults when needed. This method uses the free-living soil nematode Caenorhabditis elegans as a system to screen products for their potential anthelmintic effect against small ruminant gastrointestinal nematodes, including Haemonchus contortus. This modified method uses only liquid axenic medium, instead of agar plates inoculated with Escherichia coli, and two selective sieves to obtain adult nematodes. During screening, the use of either balanced salt solution (M-9) or distilled water resulted in averages of 99.7 (± 0.73)% and 96.36 (± 2.37)% motile adults, respectively. Adult worms tolerated DMSO, ethanol, methanol, and Tween 80 at 1% and 2%, while Labrasol (a bioenhancer with low toxicity to mammals) and Tween 20 were toxic to C. elegans at 1% and were avoided as solvents. The high availability, ease of culture, and rapid proliferation of C. elegans make it a useful screening system to test plant extracts and other phytochemical compounds to investigate their potential anthelmintic activity against parasitic nematodes.

Correlations between phenolic content and antioxidant properties in twenty-four plant species of traditional ethnoveterinary use in the Mediterranean area.

CONTEXT: Scientific information on antioxidant properties and phenolic content of less widely used plants can be useful. Therefore, the assessment of such properties remains an interesting and useful task, particularly for finding new sources for natural antioxidants, functional foods, and nutraceuticals. OBJECTIVE: As knowledge about antioxidant properties and phenolic content of many plant species used as traditional plant remedies is limited, we determined in vitro the total antioxidant activity and the phenolic content of several plant species traditionally used for ethnoveterinary practices. MATERIALS AND METHODS: For 24 extracts (70% acetone) from wild and cultivated plant species traditionally used for health care of animals we determined the Trolox equivalent antioxidant capacity (TEAC) by the two assays 1,1-diphenyl-2-picrylhydrazyl (DPPH) and the 2,2'-azinobis (3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS). The phenolic content was determined by the Folin Ciocalteu method. RESULTS: Total phenolics, calculated as gallic acid equivalent (GAE), showed variation ranging from 3.18 (Allium sativum L. (Liliaceae)) to 147.68 (Pistacia lentiscus L. (Anacardiaceae)) mgGAE/g dry weight (DW). High TEAC values corresponded to high phenolic content, while plants with low antioxidant activity exhibited low total phenolic content. The TEAC determined through each assay and total phenolic content were positively correlated, R² = 0.9152 and R² = 0.8896, respectively, for DPPH and ABTS assay. DISCUSSION AND CONCLUSION: These findings suggest that phenolic content could be used as an indicator of antioxidant properties. The results of this study encourage investigations on Mediterranean plant species as sources of antioxidants.

Pharmaco-toxicological aspects of herbal drugs used in domestic animals.

Herbal drugs are more and more used both in human and veterinary medicine to mitigate and prevent minor diseases and to support conventional medicine using allopathic drugs. Nevertheless, 'natural product' does not mean lack of adverse effects, and many people and veterinarians do not know enough about the adverse reactions that can occur following the administration of such drugs in domestic animals. Moreover, herbal products can interact with each other when administered concomitantly or can agonize or antagonize the effects of synthetic drugs administered as primary therapy. The use of non-conventional medicines (NCM) should be considered as a veterinary practise. In this paper, the herbal drugs most utilized in domestic animals, both pets and large animals, are reviewed, as their use is increasing, despite the prejudices of the academic world and some of the adverse effects and interactions that can occur in domestic animals.

Herbal usage and informant consensus in ethnoveterinary management of cattle diseases among the Kikuyus (Central Kenya).

For most smallholder farmers in Kenya conventional veterinary drugs have become very expensive and therefore unaffordable, causing them to seek low cost alternatives that are rarely documented in most ethnobiological studies. This study surveyed the utilisation of traditional herbal preparations in managing cattle ailments in Central Kenya with the aim of providing a comprehensive ethnobotanical profile and the most important plant species that may warrant scientific validation for efficacy and commercial utilisation. Using semi-structured questionnaires and detailed discussions with smallholder farmers, a total of 40 plant species in 26 families were found to be useful in traditional management of various cattle ailments in this region. Two plant families were particularly frequent in usage: Asteraceae and Lamiaceae, while the most utilised plant species were found to be Synadenium compactum N.E.Br. (Euphorbiaceae), Solanecio manii (Hook.f.) C. Jeffrey (Asteraceae) and Senna didymobotrya (Fresen.) Irwin and Barneby (Caesalpinaceae). Informant consensus was particularly high in managing anaplasmosis, East coast fever and ectoparasites. Such plant species become key target in efficacy tests and for development of commercial veterinary botanicals. The usage of some of the species is unfortunately unsustainable as some of the species are rare or endangered hence the need for conservation strategies to be undertaken.

Ethnoveterinary medicines for cattle (Bos indicus) in Bulamogi county, Uganda: plant species and mode of use.

In this paper, we present an inventory and the mode of use of plants to treat cattle. This study was carried out in Bulamogi county of Uganda, using methods consisting of semi-structured interviews employing a checklist of questions, questionnaires, direct observations, and biological inventories. Farmers employ both traditional and western medicine to treat their animals. The local people identified 33 different diseases, and had herbal treatments for nine of these. Some of the diseases mentioned by farmers indicated symptoms of diseases. Thus, the naming of diseases by local people when compared to the western veterinary medicine system, at times did not distinguish between diseases and symptoms of diseases. This is because the local disease nomenclature is based on symptoms of diseases, whereas under western veterinary science diseases are named according to aetiological information. As for traditional medicine, we report for the first time the use of 38 plant species, distributed in 37 genera and 28 families, to treat the common cattle diseases in Bulamogi. Most of these plants grow wild (76.3%), are indigenous (68.4%) and are shrubs (60.5%). The plant parts most frequently used for treating cattle are roots (37.5%) and leaves (27.5%). Medications are mostly prepared as infusions and seldom as decoctions. Topical application of some medicines is practised as well. The species used to treat cattle are also used to treat some human ailments and have some other uses as well. Most of the curative species reported here are directed at treating East Coast fever (ECF), a disease known to cause high fatalities in cattle.

Current strategies in the search for novel antiparasitic agents.

The market for antiparasitic products comprises the largest segment for sales of livestock and companion-animal healthcare agents. Despite the availability of highly effective, broad-spectrum agents, there remains a need for safer, more convenient and more environmentally friendly products that will overcome the ever-present threat of resistance development. The very high cost of discovering and developing a new drug, especially for use in livestock, is reflected in the limited number of new classes of antiparasitic agent launched on the market. New strategies are being adopted to minimise the cost of discovering potential drug candidates by maximising the chance of identifying a useful target mechanism of action and by speeding the time to discover and optimise a lead structure. These rely heavily on new technologies in target identification, screen development and lead optimisation. Examples of these will be discussed and speculation made about the possible factors that could influence the future shape of antiparasitic control.

Determination of anabolic steroids by HPLC with UV-vis-particle beam mass spectrometry.

This paper describes a general screening method for the determination of anabolic steroids in oil-based injectables, water suspensions, dietary supplements, and herbal drugs marketed in the form of capsules or tablets. The compounds are extracted into methanol, separated on a reversed-phase column with an acetonitrile-water gradient, and detected by using an ultraviolet visible (UV-vis) photodiode array detector and a particle beam mass spectrometric detector coupled in series. Identification is based on retention time, UV-vis spectra, and mass spectra. Mass spectral confirmation is accomplished by matching with a standard when available and by comparison with an electron impact mass spectra library that is either commercially available or generated in our laboratory. The tandem arrangement of the detectors provides qualitative and quantitative information from a single experiment by fully utilizing the inherent advantages associated with each detector. This method is applicable to all the anabolic steroids encountered in this laboratory so far.