Eliminación de huevos de los pequeños estróngilos en la materia fecal de los equinos y su importancia práctica en la posibilidad de tratamientos selectivos / Fecal shedding of equine small strongyle eggs and its practical importance for selective treatment

Resumen Los pequeños estróngilos son los parásitos de mayor prevalencia e importancia en los equinos de todo el mundo, y el desarrollo generalizado de la resistencia de estos nematodos a los antihelmínticos está impulsando la aplicación de tratamientos selectivos (TS) para disminuir o demorar el desarrollo de estos fenómenos. Esta estrategia se basa en conteos de huevos de nematodos en la materia fecal de todos los integrantes del grupo animal para tratar solamente aquellos que sobrepasan un determinado umbral o cut off. Este principio de TS se ve favorecido por la consistencia de los conteos de huevos por gramo de heces (hpg) en los equinos y está siendo empleada en algunos países del hemisferio norte, especialmente en los animales adultos. En la Argentina, existe una ausencia de información básica en relación a las condiciones climáticas, de pastoreo y manejo que podrían resultar en cargas parasitarias y transmisión de nematodos diferentes a las observadas en el hemisferio norte. En este contexto, el objetivo de este estudio fue caracterizar la eliminación de huevos de pequeños estróngilos en 436 equinos de diferentes edades pertenecientes a 19 establecimientos del área central de la Argentina. El 90% de los equinos mostró huevos de estróngilos en las heces y los valores de hpg fueron menores a 200 y mayores a 1.000 en el 32% y 22% de los equinos respectivamente. La distribución de los huevos de estróngilos entre individuos de todas las edades fue sobre-dispersa o agregada, con una media aritmética general de 671,59 ± 789,76 y un valor de k o parámetro de agregación inversa de 0,72 (distribución de la binomial negativa). La edad de los animales (potrillos vs adultos) y los biotipos (deportivo vs trabajo) en la categoría adultos influenciaron la magnitud de los valores del hpg (p<0,001). En esta última categoría el 35% de los animales fue responsable de la excreción de aproximadamente el 80% de los huevos eliminados en las pasturas. Si bien existen algunas diferencias con lo observado en países del hemisferio norte con poblaciones parasitarias más agregadas o sobre-dispersas, los TS para el control de los pequeños estróngilos en equinos adultos podrían también ser una estrategia válida bajo nuestras condiciones para disminuir la presión de selección sobre el genoma parasitario y permitir el desarrollo de poblaciones en refugio.

Abstract Small strongyles are the most prevalent and important parasites in grazing horses throughout the world and the widespread development of antihelminthic resistance has prompted in some countries of the northern hemisphere the application of selective treatments (ST) to reduce or delay the development of these phenomena. This strategy is based on determining strongyle fecal egg counts of all herd members and treating only those exceeding a certain threshold or cut off. However, there is an absence of basic information on horses maintained under climatic, grazing or management conditions different from those observed in the northern hemisphere. In this context, the aim of this study was to characterize the elimination of eggs of small strongyles in horses of different ages from 19 farms in the central area of Argentina. Out of a total of 436 horses, 90% showed positive strongyles egg counts and in these animals the count of eggs per gram of feces (epg) was under 200 and over 1,000 in 32% and 22% of the horses respectively. The distribution of strongyle eggs among individuals of all categories was over-dispersed or aggregated with a general epg arithmetic mean of 671.59 ± 789.76 and a value of the inverse aggregation parameter of 0.72 (negative binomial distribution). The age of the animals (foals vs. adults) and the biotypes in the adult category (sports vs. work) influenced the magnitude of the epg values (P <0.001). In the latter category, approximately 35% of the animals were responsible for the excretion of approximately 80% of the eggs in the pastures. Although some differences on magnitude and distribution of strongyle fecal egg counts were observed in countries of the northern hemisphere, ST of small strongyles in adult equines could also be a valid strategy under Argentinian conditions to reduce the selection pressure on the parasitic genome and allow the development of refugia populations.

IGF-1 abuse in sport: clinical and medico-legal aspects.

The frequent revisions of prohibited substances list established by WADA are aimed at keeping up with those drugs that, being increasingly used in medicine and in sports, play on one side a therapeutic role and, on the other, a doping role. Among the various hormone substances widely used in sports, the authors draw particular attention on IGF-1, a growth factor that is rapidly widespreading among athletes. Moreover, IGF-1 diffusion is not exclusively correlated with the doping phenomenon, being various the pathological conditions that may require the therapeutic use of this substance. As a consequence, during pathologies or medical treatments of various nature, the athletes should be informed of the IGF-1 administration because, if they are not duly apprised, they risk undergoing unjust sanctions from the competent authority. In fact, the athlete is given the possibility of communicating, before using, he is taking a medication of the WADA list for therapeutic reasons by applying for a TUE (Therapeutic Use Exemption). If this application is accepted on certified clinical grounds, the athlete will be allowed to use that substance even during sports competitions. However, the IGF-1 detection in the athlete's body shows, at the present state-of-art, two different problems: the first one has a technical character and concerns the differential diagnosis between the quantity of the physiologically produced substance and the quantity of the exogenously administrated substance; the second problem has an economic character and regards to the high cost of the analysis and, consecutively, the necessity of finding a well-equipped laboratory centre.

Medical treatment and patient decisional power: the Italian state of the art.

Therapy dissent and refusal are the situations most debated in the medical profession, that lead to various interpretations with regards to life and health, and even more in case of death risk. In the last decades, the medical treatment consent matter has become such an interesting and central issue that it has considerably modified the doctor-patient relationship. In particular, thanks to a fecund discussion on doctrine and case law, the informed consent became an autonomy act through which the patient exercises his/her personal health right. There are two international provisions that deserve to be mentioned for the matter: article no. 3 of the Charter of Fundamental Rights of the European Union, adopted in Nice on December 7, 2000, and the Oviedo Convention of April 4, 1997, which Italy passed under article no. 145, March 28, 2001. Both laws are symptomatic for the growing attention toward the person's self-determination, and for the impossibility to perform treatment in absence of personal, informed, recent and at any time revocable consent of the person concerned. Actually, these different and opposite ways of thinking correspond to a variable attitude of the case law that, in absence of a clear legislation regarding this matter, induces a deep insecurity in the physician's act on the one hand, and, does not allow the patient to totally avail him/herself of the self-determination principle on the other hand, particularly regarding the will and choices on health care.

[Newborn resuscitation: from necessity of continous practice to professional liability]. / Rianimazione neonatale:dalla necessità di formazione continua ai profili di responsabilità civile del neonatologo.

The immediately postbirth extra uterine adaptation is the most important cause of death in the first two hours of life. In all risky cases, it is necessary to effect efficient and on time techniques of newborn resuscitation, because dubitation or delay may be very dangerous for the infant. In Italy courses of equipment in newborn resuscitation are regularly performed, but an excellent level of technique can be obtained only with continuous daily practice. Then, particularly in little hospitals where it is unusually necessary to act resuscitation on a newborn, courses of simulation for medical and nursering staff would be opportune to prevent neonatal handicap and to deal with the professional liability in the best way. The Italian current jurisprudence, in fact, has slowly confined the application of 2236 article of Civil code about professional liability in particularly difficult efforts. The Italian law asserts that a professional specialist is trained to be able resolve any type of problem among those of his specialistic competence, even if technically very difficult. It should be opportune to train health staff with practical exercises, in order to obtain complete technical skills in all neonatal centers.

[What sense in cannabinoid use as regulated by Italian DM 18/04/07? Pharmacological and legal considerations]. / Quale razionale nell'impiego dei cannabinoidi previsto dal Decreto Ministeriale 18 aprile 2007? Considerazioni farmacologiche e medico-legali.

The present article relates to the Italian Ministerial Decree (DM) 18/04/2007 referring to what was established by the Financial Law 2007 on the matter of the use of drugs for the so called ''off-label'' uses. This law introduces three cannabinoid substances, with the common name of Delta 9 and Trans-delta 9 tetrahydrocannabinol and Nabilone, within the possible therapies for the treatment of ''severe pain''. The authors underline the absence of a sufficient pharmacokinetical and pharmacodynamical knowledge supporting the use of cannabinoid substances in the ''severe pain'' therapy. Further more the professional prescriber could go against judicial consequences if the drugs causes as verified the onset of collateral effects even severe that, for the scientific knowledge in possess at the present state, the authors know could take place.

[Guidelines on medically assisted reproduction: legal issues and professional liability]. / Linee Guida in materia di P.M.A.: risvolti giuridici e profili di responsabilità professionale.

The authors analyze the legal and medico-legal issues deriving from the recent Law No. 40 of February 19, 2004 concerning the Medically Assisted Reproduction. In particular, they analyze the contrasting points between the dispositions of Law No. 40/2004 and those of Law No. 194/1978 on the voluntary interruption of pregnancy, and they analyze the guidelines about the procedures and the techniques of the Medically Assisted Reproduction, issued by the Ministry of Health with D.M. of July 21, 2004. The Guidelines, as well as some sentences of several courts, lead to some reflections also about the consent and the professional liability, in particular considering the various moments of the medical action, from the first interview to the carrying out of the assisted reproduction techniques.

Anatomy of a successful imprint: analysing the recognition mechanisms of a molecularly imprinted polymer for quercetin.

This study comprises a retrospective analysis of a successful molecular imprint for quercetin with the main aim of deriving rational design strategies for more effective molecularly imprinted polymers. Hence, polymers of varying composition were synthesised and chromatographically characterised to examine the effects of monomer-template ratios. (1)H NMR analysis of the pre-polymerisation mixture yielded further information on the nature of the complexes formed prior to the polymerisation step. A direct correlation between the optimum monomer-template ratio derived from the chromatographic studies and the monomer-template ratio providing the most stable pre-polymerisation complexes observed via (1)H NMR T(1) relaxation time measurements, suggests that the formation of particularly stable pre-polymerisation complexes is responsible for an increased formation of selective binding sites during the polymerisation step. Furthermore, physical aspects of the polymerisation, such as the MIP surface area and macroscopic phase partitioning of the mixture during the polymerisation are investigated. The observed effects and their analytical assessment offer insight into the mechanisms governing MIP selectivity at a molecular level.

Towards an GMO discipline: ethical remarks.

The present technological transformation calls for a new kind of vet: he has to accomplish his own institutional tasks bearing in mind the definition of a "good practice" code and also performing educational and informative tasks in zootechnology in order to ensure the animal well-being. Therefore, his contribution can be relevant in developing and interpreting laws related to the protection of animal welfare and further considerations on the ethical and deontological thought, concerning the technological use of animals. The vet's role is also fundamental in the following areas: further study of bioethical issues related to zootechnology and the professional growth of whoever is engaged, in different ways, in productive activities and research on animals.

Advances in the analysis of mycotoxins and its quality assurance.

This article covers the latest activities in mycotoxin analysis and the advances of its respective quality assurance. The majority of mycotoxin analyses carried out in the laboratories is still based on physicochemical methods, which are continually improved. For example, immunoaffinity columns and multifunctional clean-up columns have become of increasing importance and in some areas of mycotoxin analysis they have more or less displaced conventional liquid-liquid partitioning or column chromatography during clean-up. The need for rapid yes/no decisions on the other hand has led to a number of new screening methods. In particular, rapid and easy-to-use test kits based on immunoanalytical principles or the generation of artificial macromolecular receptors employed in molecularly imprinted polymers (MIPs) have made good progress. Further research in mycotoxin analysis is pursued in the field of biosensors and also the potential of infrared spectroscopic techniques as screening method has been demonstrated. In the area of multi mycotoxin analysis the most promising development was observed in mass spectrometry. At the same time, several interlaboratory studies in the field of mycotoxin analysis revealed problems proven by high between laboratory standard deviation and non-traceable results. This not only shows the necessity of reliable methods and well defined performance characteristics but also the need for appropriate calibrants of defined concentration and stated purity. A certified zearalenone (ZON) calibrant is already available and a certified calibrant containing various trichothecenes is currently under development. (Certified) reference materials are available for aflatoxins in a number of commodities, ochratoxin A (OTA) in wheat, deoxynivalenol (DON) in maize and wheat, and ZON in maize. With these measures important steps towards traceability of results in mycotoxin analysis have been achieved.

Voluntary interruption of pregnancy in Europe: medico-legal issues and ethical approach to the regulation.

The legislative criteria of the various European countries, as far as voluntary interruption of pregnancy is concerned, differ in the levels of severity employed by the law of the single nations. The authors analyze the legislation of some of European countries and, as a result, they identify countries that may be defined as more tolerant and others where the law is more restrictive. Italy is to be dealt with separately, because its legislation as regards this subject presents a greater interpretative ambiguity. In the end they analyze religious attitudes underlying the laws on voluntary interruption of pregnancy, because of the suspicion that the teachings of both Catholic and Protestant theology have influenced the differences in the quality of the law in force as regards termination of pregnancy is certainly well-grounded, and on the other hand the philosophy of law certainly owes its meditation both to theology and to the teachings of the Catholic Church.

Towards the rational development of molecularly imprinted polymers: 1H NMR studies on hydrophobicity and ion-pair interactions as driving forces for selectivity.

The preparation of molecularly imprinted polymers (MIP) based on non-covalent interactions has become a widely used technique for creating highly specific sorbent materials predominantly used in separation chemistry. A crucial factor in a successful imprinting protocol is the optimisation of the template/functional monomer interaction in the pre-polymerisation mixture, eventually leading to a maximum of high-affinity binding sites in the resulting polymer matrix. In order to develop more efficient preparation technologies for imprinted polymers, two separate pre-polymerisation complexes were investigated by NMR spectroscopic techniques in order to identify the types of interactions occurring in the pre-polymerisation mixture, and their implications for the subsequently formed imprinted polymer. In particular, hydrophobic effects have been followed by NMR spectroscopy and their contribution to the selectivity of the resulting MIP has been investigated. The 2,4-D imprint system is used as an example to fundamentally study whether observations at the pre-polymerisation stage correlate with properties of the finally prepared MIP, and which parameters govern success of an imprinting protocol.

Molecular imprinting and solid phase extraction of flavonoid compounds.

Molecularly imprinted polymers (MIPs) for quercetin have been successfully prepared by a thermal polymerization method using 4-vinylpyridine (4-VP) and ethylene glycol dimethacrylate (EDMA) as functional monomer and cross-linker, respectively. The obtained molecularly imprinted polymers were evaluated by HPLC using organic eluents, with respect to their selective recognition properties for quercetin and related compounds of the flavonoid class. Two equivalent control polymers, a blank polymer and a polymer imprinted with a structural analogous template, were synthesized, in order to confirm the obtained results. Furthermore, preliminary experiments confirm the applicability of the prepared MIPs for solid phase extraction (SPE), as rapid and facile clean-up of wine samples for HPLC analysis is an envisaged field of application. The successful preparation of molecularly imprinted polymers for flavones provides an innovative opportunity for the development of advanced separation materials, with applications in the field of wine and fermentation analysis.

"Single step" PCR with a sensitivity similar to nested PCR for the detection of hepatitis C virus RNA.

OBJECTIVE: Evaluation of the performance of different HCV PCR detection systems for HCV RNA: A nested PCR, considered the reference assay, was compared with two single-step methods (ss-PCR): the first is based on the detection of PCR products by liquid hybridization with a 32P end-labelled probe (isotopic ss-PCR), while the second assay is a colorimetric method (colorimetric ss-PCR) using microwell plate hybridization with a specific nucleic acid probe (Amplicor HCV PCR, Roche Diagnostics Systems). METHODS: Sera from 56 patients with suspected hepatitis C infection based on reactive serology or altered liver parameters, and sera from 15 blood donors were tested for HCV RNA: After RNA extraction, the synthesized HCV cDNA was amplified in parallel using isotopic ss-PCR, colorimetric ss-PCR and nested PCR. The products were detected by autoradiography, color development and ethidium bromide fluorescence, respectively. RESULTS: In order to assess the analytical sensitivity of ss-PCR versus that of nested of PCR, experiments included serial dilutions of positive control samples. Results showed that both methods had an extinction signal at the 1:512 dilution. A comparative analysis of 71 clinical sera samples was obtained using the three protocols and the results clearly documented 100% concordance. CONCLUSIONS: Single step PCR methods for HCV RNA have a sensitivity equal to that of nested PCR and appear more suitable for diagnostic applications. Ss-PCR is safer than nested PCR in terms of both specificity and contamination problems. In particular, the Roche Amplicor HCV PCR assay minimizes sample exposure and management problems.

Detection of HIV-1 proviral sequences in lymphocytes using a qualitative polymerase chain reaction assay.

The performance and clinical relevance of a qualitative PCR-based assay for the detection of HIV-1 DNA sequences in peripheral blood mononuclear cells (PBMCs) was evaluated by two different laboratories. Four hundred and one samples were obtained from 397 individuals from different risk populations. All blood donors tested had negative results; positive signals were obtained from all infected patients. HIV-1 DNA was detected in 3 of 17 infants born to seropositive mothers; Western blot indeterminate blood donors and exposed health-care workers had negative results. Our results demonstrate that this PCR assay provides both sensitive and specific results and is suitable for testing large numbers of samples and for rapid identification of HIV-1 infection.