Comparative activity of some veterinary pharmaceutical products in swine dysentery

Authors: Romeo T. Cristina1, Roberto Villa2, Olimpia-Ines Lupu (Paută)1, Alexandru Doma1

Affiliation: 1 USAMVB Timisoara, Facultatea de Medicina Veterinara                                                                                2 Universita di Milano, Facolta di Medicina Veterinaria

One of the diseases that cause great damage in the pig breeding farms, Brachyspyra swine dysentery has a special importance because involve great losses both by growth failure and high mortality and significant costs to the prevention and control. The study was conducted in a swine farm in Timis County, where the disease was originally identified in autumn of year 2009. Cumulative adverse conditions have affected the rate of morbidity and mortality. The most common clinical developments in the unit were usually acute and subacute. Research included: a comparative study of the effectiveness of tiamulin fumarate, hydrogenated and dimetridazole powder in sick acute and subacute forms in piglets, at the weaning period; the comparative study of efficiency in therapy of soluble tiamulin (the tiamulin fumarate-hydrogenated) and of the injection of piglets effective and the study of growth indicators: average daily gain.
Experiment I
Group I consisted in 15 piglets with an average weight per lot of 7.60 kg/head (batch weight: 114kg) received p.o. dimetridazole insoluble powder (Romvac Bucharest), mixed in feed, for 5 days dose of 25mg kg.bw. During the observation, two mortalities were recorded, representing 13.33% of the treated lot. Other animals from the group were clinically cured, the signs of the disease gradually disappeared, and the temperature returned to normal enteric symptoms. The observation period after the 21 days of experiment, the batch weight was 114.2 kg, corresponding to growth registered 95.5 g/head/day.
Group II, of 15 pigs with enteritis clinical signs, have had an average weight per lot of 7.48 kg/head, and he has undergone treatment with tiamulin product (Novartis Animal Health) soluble granules, in drinking water. Treatments were performed for 5 consecutive days at a dose of 60 mg/litre of drinking water. In this group, diarrhoea symptoms decreased and general status of the animals gradually improved, no mortality was recorded five days after the treatment, no longer signs of illness being observed, all animals became normothermic. Weight gain recorded on our observation period was 29.3 kg/ lot which correspond to average daily gain of 162.7 g/day/animal.
Group III, considered as control group, was at the beginning of the observation period, established with the average weight per lot, of 7.88 kg/head. In this series evolved enteritis with clear clinical manifestations (acute and subacute forms) and during the experiment did not receive any treatment. Within this lot, in the studied period have died five piglets which represent 33.33% of the group. Consequently, to avoid the unnecessary losses, in the 21 day of experiment, the 10 remaining piglets were orally treated with tiamulin (60 mg/litre drinking water). Subsequent observations have shown gradual improvement and clinical remission to the remaining piglets. In the control group, practically can’t talk about a weight gain. After the 21 day average daily gain for animals remaining batch was 45.1 g/day/animal.
Experiment II
The groups were selected from age group 35-45 days, which are each, composed of 12 heads per group.    Group I was treated with soluble granules tiamulin concentration of 45 mg of active substance per litre of drinking water. Treatment was performed over a period of 5 days. During the experiment, within this group there was one death case representing 8.33%. The remaining 12 piglets were cured within two weeks. Initial weight was of 92.9 kg / group, and after 21 days of experiment, it reached at 98.9 kg, corresponding to an average daily gain of 101.5 g/head/day. Although, there was loss of a pig, average daily gain of piglets remaining was considered positive, the group recorded an increase of 6 kg in the 21 days of study.
Group II received in drinking water, soluble tiamulin at concentration of 60 mg / litre for 5 consecutive days. In this group there was no mortality, all 12 piglets to heal within 10 days (cure rate 100%). The final weight for this group was of 117.6 kg (a plus of 24.9 kg), the recorded growth being of 172.8 g/head/day.
Group III was treated the injectable product Tiamutin 200 (1ml / 20kg.bw. intramuscularly for 4 days. Registered losses were 2 pigs, the remaining 10 piglets healing within 10 to 14 days. If this group, because of the recorded mortality (average daily gain 79.1 g/head/day), have not economic efficiency.
Group IV – control has not received any treatment within 7 days of the experiment.
During this time four piglets died (all with forms of hemorrhagic enteritis) and accounted for a proportion of 33.3% of the lot. After those seven days, in the aim to reduce the unnecessary losses, the remaining piglets were treated with tiamulin soluble dose to 90 mg / l for 5 consecutive days. Note that all the remaining 8 animals treated were cured. The economic indicators showed clearly the weight declining. At the end of the experiment, there was a loss of 26 kg and four dead piglets. In conclusion, the comparative use in therapy of tiamulin and dimetridazole, preparations having effect in swine’s dysentery, have shown that, both have certain therapeutic effects, but tiamulin far proved to be most effective. The results come to confirm other studies showing superior efficacy of tiamulins as preventive and curative choice in swine dysentery with Brachyspira hyodysenteriae.

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Veterinary medicinal products for the bees – the current situation and future strategies – an important topic discussed at European level

Authors: Alina Karina Draghici, Anca Bitoiu, Lollita Taban                                                               Affiliation: Institutul pentru Controlul Produselor Biologice si Medicamentelor de Uz Veterinar, Bucuresti

To analyze the current situation and future issues relating to health and treatment options bees, bee breeders in Europe, agencies and drug manufacturers in Europe have held several meetings. One of these took place last year in December at the EMEA (European Medicines Agency) in London, United Kingdom. The purpose of this meeting was to consider the current situation of pathology in bees to identify the most common diseases found in this species, identification and lack of treatment options for some diseases, and identifying solutions to improve the situation.

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Development and validation of a RP- HPLC method for the quantittation studies of metronidazole and furazolidone from product Enteroguard M

Authors: Elena Gabriela Oltean1, Milea B.2

Affiliation: 1 Romvac Company SA,                                                                                                                                                                          2 Proanalysis Systems SRL

An isocratic high-performance liquid chromatography (HPLC) procedure was developed for quantitative determination of metronidazole and furazolidone in tablet dosage forms of ENTEROGUARD M. HPLC separation was carried out by reversed phase chromatography Kromasil C18 (250 mm x 4,6 mm i.d.; 5 +m particle size), held at 25°C. The mobile phase consisted of methanol/ 0,1% phosphoric acid aq. (20/ 80 v/ v), run at flow rate of 1 mL/ min and with UV detection at 317 nm. Method validation investigated parameters such as linearity (r2 = 0. 9999), range, precision, accuracy and specificity. The described method can be successfully applied for the analysis of ENTEROGUARD M tablets.

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Analytical method (HPLC), validation used for identification and assay of the pharmaceutical active ingredient, Tylosin tartrate for veterinary use and its finite product Tilodem 50, hydrosoluble powder

Author: Maria Neagu                                                                                                                   Affiliation: SC DELOS IMPEX ’96 SRL

In SC DELOS IMPEX ’96 SRL the quality of the active pharmaceutical ingredient (API) for the finite product Tilodem 50 – hydrosoluble powder was accomplished in the respect of last European Pharmacopoeia. The method for analysis used in this purpose was the compendial method „Tylosin tartrate for veterinary use” in EurPh. in vigour edition and represent a variant developed and validation „in house”. The parameters which was included in the methodology validation for chromatographic method are the followings: Selectivity, Linearity, Linearity range, Detection and Quantification limits, Precision, Repeatability (intra day), Inter-Day Reproductibility, Accuracy, Robustness, Solutions’ stability and System suitability. According to the European Pharmacopoeia, the active pharmaceutical ingredient is consistent, in terms of quality, if it contains Tylosin A – minimum 80% and the amount of Tylosin A, B, C, D, at minimum 95% Identification and determination of each component separately (Tylosin A, B, C, D) is possible by chromatographic separation – HPLC. Validation of analytical methods is presented below.

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Contributions to the humoral immunological profile in broiler chickens treated with 50% Amoxidem

Authors: Turcu D1., Mariana Oporanu1, Condur D1., Grigorescu P1., Roman M2.

Affiliation: 1 Facultatea de Medicinã Veterinarã Spiru Haret, Bucuresti                                                                              2 S.C. Delos Impex 96 S.R.L.

Investigation on the effect of Amoxidem 50% humoral immunological effectors (protein fractions, lysozyme, total protein) in broilers was conducted. There were two groups of chickens tested: control group A and group B treated with Amoxidem 50%. Blood sampling was performed 7 days after treatment. Electrophoretic pattern of serum protein fractions in the experimental group showed a significant increase in the concentration of globulin fractions α1, α2 globulin and γ globulin and a decrease in the A/G ratio. Mean IgG level increased significantly (p˂0.001) in group B compared to group A. The serum’s lysozyme concentration showed a significant increase in statistical terms (p˂0.01) in group B compared with group A.

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LC-MS/MS Analysis of albendazole and its metabolites in animal tissues

Authors: Ana Csuma, Ana Cismileanu                                                                                           Affiliation: S.N. Institutul Pasteur S.A., Bucuresti

Treating the animals with veterinary medicines raise the issue of residues that can persist in their edible tissues. In order to establish the withdrawal time necessary for depletion of the residues to sufficiently low concentration not to affect human health, biological tests performed on animals implicitly involve the use of sensitive and reliable analytical methods for residues determination. The aim of this work was to establish and validate a sensitive and reliable method for simultaneous determination of albendazole and its metabolites, albendazole sulfoxide, albendazole sulfone and 2-aminoalbendazole sulfone in animal tissues by LC/MS/MS. The method involves acid hydrolysis with 6N HCl in order to release most residues, in particular the bound metabolite 2-amino-albendazole sulfone especially from liver, followed by extraction with ethyl acetate and solid phase clean up of the extract on a C18 SPE cartridge. The liquid chromatographic separation was achieved on a XTerra MS C18 column (10 cm x 2,1 mm, 3,5 ìm), with gradient elution of 0,1% formic acid – methanol. Detection was performed by mass spectrometry in ESI+ mode. The limits of quantification were lower than 4 ìg/ml for each component.The correlation coefficients (R2) of the calibration curves (in the range from 0,01 ìg/ml to 0,5 ìg/ml) were higher then 0,9935. The relative standard deviations of repetability on samples naturaly contaminated from treated animals were between 5,75% and 11,6%. Recoveries from fortified muscles in the range of 10 ìg/kg to 100 ìg/kg were between 70,2% and 88% with relative standard deviation of 5,4% – 12,2%. For liver fortified in the range of 100 ìg/kg to 1000 ìg/kg recoveries between 70,3% and 83,2% were obtained with relative standard deviation of 5,7 %-11%.

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Contamination of food with residues of antibiotics in the sulphonamide class, a risk that can be avoided

Autori: Carmen Lidia Chițescu1, Anca Nicolau2

1 S.C. Pasteur, Filiala Filipesti,                                                                                                                     2 Universitatea Dunarea de Jos, Galati

Sulfadimethoxine, sulfamethoxazole, sulfaquinoxaline and sulfadiazine are the most common used sulfonamides in veterinary practice. The recommended withdrawal periods if not observed before slaughtering of the medicated animals, the products may obtain from such animals may be contaminated with residue. The interest in having reliable methods able to detect low amounts of sulfonamides in food is very actual. In this study, a multiresidue analysis was performed to simultaneously determine those four sulfonamides in chicken muscle tissue by the Waters LC. Criteria of validation: specificity, accuracy, precision, limit of detection, limit of quantification, and linearity, according to the European Commission Decision 2002/657/EC, show that the method can detect different kinds of sulfonamides within one run, without mass spectrometry analyses, or Fluor metric derivatization of analyts. The method is accurate, simple, economical in both time and cost, capable of detecting sulfonamides residues below the maximum residue limits (MRL) and easy to perform to routine samples, in normal condition of laboratory. The sulfonamides were extracted with acetonitrile and acetone and dichloromethane. N-hexane was added for defeating the sample. Separation was carried out on a Zorbax SB- C18 analytical column, using as mobile phase a mixture of 75:25 = di-natrium-hydrogenphosphat solution 6 g/1000 ml (pH = 8.5) : methanol. The detection wavelength was set at: 254 nm Calibration graphs were linear with very good correlation coefficients in the concentration range from 0.320 to 1.5ìg /mL. The limits of quantification (LOQ) for the sulfonamides were in the range of 6.6–0.34 ìg /kg. The recovery for spiked chicken muscle with 50–150 ìg/kg ranged more than 70%. The relative standard deviation (Reds) of the sulfonamides for six measurements at 50 go/kg, 100 ìg /kg and 150 ìg /kg was less then 15%. The applicability of the method to the analysis of chicken muscle tissue was demonstrated.

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Theoretical elements of veterinary therapeutics

Autori: Romeo T. Cristina1, Ivo Schmerold2, Cosmin I. Tuns1

Affiliation: 1 Facultatea de Medicină Veterinară Timișoara,                                                                                               2 Veterinarmedizinische Universitat Wien

The present essay describes, logically and in a synthetic way, the main elements of veterinary therapeutics. The first part refers to the concept of dose and the main factors affecting its establishment, presenting the elements related to: weight and body surface area, the therapeutic range, animal’s size, genetic factors, specific variations, digestive tract anatomy, factors related to age, sex, pregnancy, time of administration, tolerance and
intolerance to medications, and factors that may influence response to the medication (diminished/amplified response, dose administrations toxicity, etc.). Following are revealed the key factors that determine the administrations’, frequency like: drugs clearance, drugs’ concentration, administration rate, repeated-dose effect and the frequency establishing, etc. There are no missing also the elements that are describing: the plateau effect, zero-order kinetics, drug residues effect, the dose-effect relationship, the first-pass effect, latency and intensity, the risk-benefit ratio in the veterinary therapy, to full understanding of the concepts by the vet practicians.

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