Antibodies elicited by the CaniLeish® vaccine: long-term clinical follow-up study of dogs in Spain.

The prevention of canine leishmaniosis in healthy dogs requires a multimodal approach combining repellents with an effective vaccine. A vaccine that modulates the cell-mediated immune response against the protozoan has been available in Europe since 2012 (CaniLeish®, Virbac, France). The aim of the present study was to monitor dogs vaccinated with CaniLeish® to examine the kinetics of the antibody response and the safety and tolerance of CaniLeish®. Dogs vaccinated with CaniLeish® were monitored for 12 months. In follow-up visits at baseline (primovaccination or annual booster) (Visit 1, V1), and 1 (V2), 4 (V3), 8 (V4) and 12 (V5) months later, we examined antibody response kinetics using two serology techniques (IFAT and Speed Leish K™). Tolerance to CaniLeish® and its safety were also monitored. Anti-L. infantum IgG antibodies were determined in 242 dogs (125 dogs after primovaccination (Group P) and 117 dogs after booster vaccination (Group B). In addition, 46, 22 and 19 dogs were followed for 2, 3 and 4 years, respectively. At baseline, 100% of dogs in Group P returned negative IFAT and Speed Leish K™ test results while 9.4% (11/117) in Group B tested IFAT positive though Speed Leish K™ negative. In subsequent visits, seropositivity was detected by IFAT in 31.2% (Group P) and 41% (Group B) of the dogs in V2; 16.8% (Group P) and 10.2% (Group B) in V3; 6.4% (Group P) and 8.5% (Group B) in V4; and 3.2% (Group P) and 5.9% (Group B) in V5. All dogs tested Speed Leish K™ negative except two, in which it was later confirmed by molecular testing that they were not infected. Adverse events that could be associated with the vaccine were detected in 20 out of 314 dogs (6.4%). The good clinical status of all dogs was confirmed in an exhaustive clinical exam and haemato-biochemical profile. The Canileish® vaccine was well-tolerated with exceptions that did not appear to be related to age, sex, race or size of vaccinated dogs. Anti-L. infantum antibodies were detected by IFAT in 31.9-40.3% of the dogs 1 month after vaccination, and these antibodies could still be detected in 3.2% of the dogs 1 year later. This means that veterinarians need to use other tools (eg. PCR) to correctly diagnose seropositive dogs.

Flea species infesting dogs in Spain: updated spatial and seasonal distribution patterns.

This entomological survey examines the spatial and seasonal distribution patterns of flea species infesting dogs in Spain. Bioclimatic zones covering broad climate and vegetation ranges were surveyed according to size. In a cross-sectional spatial survey carried out from late May 2013 to mid-July 2015, 1084 dogs from 42 different locations were examined. A total of 3032 fleas were collected and identified as belonging to the following species: Ctenocephalides felis (Siphonaptera: Pulicidae) (81.7%, 2476 fleas); Ctenocephalides canis (11.4%, 347 fleas); Pulex irritans (Siphonaptera: Pulicidae) (6.9%, 208 fleas), and Echidnophaga gallinacea (Siphonaptera: Pulicidae) (0.03%, one flea). Variables observed to have effects on flea abundance were animal weight, sex, length of hair and habitat. In the seasonal survey conducted from June 2014 to June 2015, 1014 fleas were collected from 239 dogs at 30 veterinary practices across Spain. Peaks in C. felis abundance were observed in early summer and late autumn, whereas high numbers of P. irritans and C. canis were recorded in autumn. Numbers of fleas detected in winter were low overall. Based on these findings, the present study updates the spatial and seasonal distributions of flea species in Spain and assesses the impacts of host and habitat variables on flea infestation.

Effect of two treatments on changes in serum acute phase protein concentrations in dogs with clinical leishmaniosis.

The objective of this study was to compare changes in serum concentrations of acute phase proteins (APPs) and paraoxanase (PON-1) in response to two treatments in dogs with leishmaniosis (CanL). For this purpose, 20 dogs with CanL were assigned to two treatment groups: antimonial plus allopurinol (Group G, n=12) and miltefosine plus allopurinol (Group M, n=8). Serum concentrations of PON-1 and APPs including C-reactive protein, haptoglobin (Hp), ferritin (Ft) and albumin were monitored over a period of 3 months after treatment. At the beginning of the study (day 0), most of the dogs had APP abnormalities. None of the variables differed significantly between groups in the first or subsequent visits. There was a significantly higher reduction in serum Ft in Group G than in Group M from day 0 to day 30 (P=0.0085), and also from day 0 to day 90 (P=0.0214). There was a higher increase in serum PON-1 in Group G than in than Group M from day 0 to day 30 (P=0.0039), and also from day 0 to day 90 (P=0.0404). This is the first report of APPs in dogs with natural clinical leishmaniosis treated with miltefosine. There was faster resolution of serum APP concentrations in dogs treated with antimonials (P<0.05).

Implications of zoonotic and vector-borne parasites to free-roaming cats in central Spain.

Cats are definitive hosts and reservoirs for several parasites, some of which are responsible for serious zoonotic diseases. We conducted a case-control study of data from a trap-neuter-return (TNR) programme (years 2014-2017) designed to examine the prevalence of zoonotic parasites in free-roaming cats living in urban areas of central Spain. In the animal population tested (n = 263), we detected a 29.2% prevalence of endoparasites, including high rates of cestodes (12.9%) and Toxocara cati (11.7%). While faecal samples showed no Toxoplasma gondii oocysts, the seroprevalence of T. gondii infection was 24.2%. Antibodies to Leishmania infantum were detected in 4.8% of the animals, though all skin and blood samples analyzed were PCR negative for this parasite. Ectoparasites (ticks and fleas) were found in 4.6% of the cat population, and 10.6% of the cats were detected with Otodectes cynotis. Finally, 6.3% and 7.9% cats tested positive for feline leukaemia virus and feline immunodeficiency virus, respectively. Our study provides useful information for animal-welfare and public-health, as the parasites detected can affect native wild animals through predation, competition and disease transmission. Our detection of zoonotic parasites such as L. infantum, T. gondii, T. cati, Giardia duodenalis and several ectoparasites prompts an urgent need for health control measures in stray cats.

High-performance liquid chromatography determination of Zn-bacitracin in animal feed by post-column derivatization and fluorescence detection.

A sensitive and selective method is presented for the determination of Zn-Bacitracin in adulterated animal feed by reversed-phase ion-pair high-performance liquid chromatography and post-column derivatization with o-phthalaldehyde prior to fluorescence detection. The calibration function was estimated to be between 8.0 and 65.0 mg l(-1) of Zn-BC. The detection and quantification limits of the chromatographic method were 2.5 and 7.5 mg 1(-1), respectively. Using the extraction procedure of Zn-Bacitracin from the feedstuff that we recently proposed and applying this new chromatographic method, it was possible to detect this antibiotic at levels below 5 mg kg(-1) in different kinds of feedstuffs with a standard deviation less than 6.0%.

Determination of five nitroimidazoles in water by liquid chromatography-mass spectrometry.

A rapid liquid chromatography-mass spectrometric method for the determination of five nitroimidazoles in water from different sources is described. The extraction procedure was based on HLB solid-phase extraction with acetonitrile followed by an evaporation step. Ternidazol, another nitroimidazole, was used as the internal standard. The liquid chromatographic separation was made on a C18 bonded silica column applying a gradient with an ammonium acetate buffer solution and acetonitrile. Following electrospray ionisation, the protonated molecular ions [M+H]+ were obtained. Quantification for each nitroimidazole was carried out by monitoring its molecular ion. Calibration functions, quantification and detection limits, intra- and inter-day reproducibility and accuracy were estimated. For the confirmatory assay, several fragment ions from each nitroimidazole were obtained and monitored. The method was applied successfully to determine and identify nitroimidazoles in water from different sources at a level of 0.2 microg l(-1).

Determination of iron at ng/ml level by solid phase spectrophotometry after preconcentration on cation exchange filters.

A cationic exchanger paper is used to retain analytes in solution and, after drying, to analyze directly by measuring the UV-Vis absorbance of the paper. The method was applied to determination of iron using its known 1,10-phenanthroline complex. Using 100 ml of sample the applicable concentration range was between 1.0 and 10.0 ng/ml with a detection limit of 0.2 ng/ml and a RSD around 2%. The method was applied to determination of iron total in snow, human serum and wine.

Density functional study of layering at liquid surfaces.

We analyze the density profiles for liquid-vapor interfaces within two density functional (DF) approximations, applied to simple fluid models which have low ratios between their triple and critical temperatures. The observation of layering structures at low T is discussed in relation with the Fisher-Widom line for each model. Although we find no apparent correlation between the amplitude of the oscillatory density decay mode and the approach to T(FW) , that temperature sets a threshold for the generation of nonmonotonic structures within a fixed distance of the interface. The rapid decay of the oscillatory mode amplitude with T may be interpreted as a result of the capillary wave (CW) damping of strongly structured intrinsic density profiles. The layering in the presence of gravitylike external fields indicate that the effective transverse size which might be built in the DF approximations is around 10+/-2 molecular diameters; however, that interpretation has to allow for an effectively reduced damping exponent, i.e., an effective surface tension for the CW Hamiltonian which is larger than the value obtained directly from the DF grand-potential minimization.

Brucellosis infection presenting with cholestasis.

Brucellosis is a relatively common disease in the Mediterranean area and may present with prolonged fever without focus, however it remains an important diagnostic challenge to most pediatricians. We report the case of a 10-year-old male patient who presented with fever without a focus of 10-day duration, hepatomegaly, ascites, a small elevation in transaminases and acute-phase reactants indicating cholestasis, leukopenia, and thrombocytopenia. Imaging tests showed many small, rounded, hypodense focal lesions in the liver and spleen. After eliminating a wide range of diseases, positive results for the Rose Bengal test and indirect immunofluorescence assay for Brucella melitensis made it possible to establish a diagnosis of hepatosplenic brucellosis. A review of the family history revealed direct contact with farm animals. The patient made good progress on treatment with doxycycline and streptomycin, with complete resolution of both clinical symptoms and imaging signs. The prevalence of brucellosis is gradually increasing, mainly due to migratory movements. It should always be eliminated as a source of unknown fever in endemic areas and should also be taken into account in other geographical areas where it is not common whenever a patient presents with prolonged fever and unspecific symptoms. Standard therapy is highly effective, even in relapse cases, and early diagnosis leads to a rapid recovery and favorable outcome. The unusual presentation in the case reported here reminds us that it is possible to encounter a Brucella infection in a case of fever without a focus, irrespective of the existence of a clear epidemiological history, which is very often omitted by the family. All differential diagnostic protocols for fever without a focus should include a diagnostic test for Brucella in order to achieve early detection of the disease and initiate therapy promptly.

Critical analysis of the density functional theory prediction of enhanced capillary waves.

We present a critical analysis of the density functional description for capillary wave fluctuations on free liquid surfaces. The proposal made by Mecke and Dietrich, [Phys. Rev. E 59, 6766 (1999)10.1103/PhysRevE.59.6766], to obtain the effective wave vector dependent surface tension, and their prediction of an enhanced regime of capillary waves at mesoscopic scales, has had a large impact including claims of experimental observation [Fradin, Nature (London) 403, 871 (2000)10.1038/35002533; Mora, Phys. Rev. Lett. 90, 216101 (2003)10.1103/PhysRevLett.90.216101]. Our analysis shows that there is a qualitative problem in the convergence of the low q expansion used for that prediction, and that the assumed link between the equilibrium density functional description of the liquid surface and its capillary wave fluctuations leads always to the unphysical decrease of the surface tension for large wave vectors.

Improvement in LC-MS calibration by addition of fragment signals.

Liquid chromatography-mass spectrometry has become a powerful analytical tool, with high selectivity and sensitivity. Usually in this technique, the calibration function is estimated from the molecular peak signal. This report describes the improvement in sensitivity when the signals from several fragments in addition to the molecular peak are used to establish the calibration function. The influence of the dwell time has also been analysed as an important instrumental parameter that influences the signal range, and consequently, the sensitivity. The calibration function obtained by adding fragment signals was used to estimate the instrumental detection limit using three different procedures, comparing and discussing the results obtained.

Utilidad de los marcadores tumorales

Se describen las características de los diferentes grupos de marcadores tumorales sericos, clasificados según su origen y propiedades de mayor o menor especificidad y finalmente se comentan los casos en que estas sustancias pueden ser de ayuda en la práctica clínica.

Sarcomas de partes blandas de extremidades: experiencia de 20 años / Sarcomas of soft parts of extremities: experience of 20 years

Estudia 88 casos de sarcomas de partes blandas representados por el 1.14xciento de todos los tumores diagnosticados en el Servicio de Oncología del HCAM desde 1973 a 1993. La presentación en extremidades ocurrió en el 52xciento de estas neoplasias. La frecuencia por edades fue mayor entre los 30 a 40 años y sobre los 60 años, observándose un incremento en el número de casos por año en los últimos 5 años dando una incidencia anual de 3.6 casos. La relación hombre/mujer fue de 24 vs 20 y la tasa de comparación entre extremidades inferiores superiores fue de 2.6 a 1. El fibrosarcoma fue el tipo histológico más común (25xciento) seguidos del liposarcoma y rabdomisarcoma (23xciento c/u). El 43xciento correspondió al estadío I, el 48xciento a estadío II y el 9xciento a estadío III. La tasa de recurrencia global fue del 64xciento, siendo del 9xciento para las locales y de 55xciento para las distales. Las modalidades de combinación representaron las terapias más utilizadas (80xciento) y la sobrevida global a 5 años fue del 53xciento...

Catéteres venosos centrales permanentes: uso en quimioterapia, complicaciones y morbilidad

El propósito de revisar la morbilidad de los catéteres colocados en el servicio de oncología para tratamiento de quimioterapia. Entre noviembre de 1992 y julio de 1994 se colocaron 100 catéteres venosos centrales (CVC) en 98 pacientes, que presentaban alguna de las siguientes características: 1.- en tratamiento de quimioterapia (QT) sin venas periféricas adecuadas, 2.- pacientes en QT de larga duración, 3.- Pacientes pediátricos o 4.- Pacientes obesos. La colocación se la realizó con adecuadas medidas de asepsia y bajo control fluoroscópico. El rango de edad va de los 3 a los 74 años, con una media de 45 años. Se colocó a 34 hombres y 64 mujeres con diferentes tipos de neoplasias. El sitio fue en subclavia en 92 pctes., en Basilica en 5 pcts., en yugular en un pcte. y fallidos en 2 casos. De los 100 CVC, 18 fueron subcutáneos (implantofix) y 80 externos con tunelización en la región pectoral. La principal complicación fue el dolor localizado en 13 casos e infecciones locales en 10 casos, hubo un caso de neumotorax y dos punciones a la arteria subclavia, todas las complicaciones se resolvieron adecuadamente, no se produjo ningún fallecimiento a causa del procedimiento. La duración media del CVC fue de cuatro meses y medio. Se comprobó la utilidad del uso de los CVC en el paciente que recibe QT, facilita su administración y la tasa de complicaciones presentadas es similar a otros estudios públicados.