Hemolysis, icterus and lipemia interfere with the determination of two oxidative stress biomarkers in canine serum.

BACKGROUND: Oxidative stress has been proven to play a role in numerous human and canine diseases. Among the biomarkers of oxidative stress, Thiobarbituric Acid Reactive Substances (TBARS) and Total Antioxidant Status (TAS) are two of the most widely used. Preanalytical factors are crucial for obtaining accurate results in these assays. Hemolysis, icterus and lipemia (HIL) are common sources of preanalytical errors in the laboratory; however, limited information is available regarding the considerations for canine specimens. Therefore, the objective of this study was to evaluate the potential interferences of HIL in the determination of TBARS and TAS in canine serum. METHODS: Solutions of pooled canine serum samples were prepared by adding increasing concentrations of hemolysate, bilirubin and a synthetic lipid emulsion. TBARS and TAS were determined, and biases from the control value caused by the interfering substances were calculated. RESULTS: Hemolysis, icterus and lipemia induced significant interferences on TBARS and TAS, albeit to varying degrees depending on the specific biomarker and interfering substance. TBARS appeared to be more susceptible to interferences in this study. Slight hemolysis, moderate icterus and slight lipemia caused notable deviations in TBARS values, surpassing the acceptable threshold for interference. TAS assay was also affected by HIL, although to a lesser extent compared to TBARS. Significant biases from TAS control value were observed when icterus was moderate, and when hemolysis and lipemia were more pronounced. CONCLUSIONS: In light of our results, we conclude that hemolyzed, icteric and lipemic specimens are not suitable for TBARS and TAS determination in canine serum. Our findings hold considerable practical utility, as a simple visual inspection would be sufficient for identifying and excluding such specimens.

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.

Short term impacts of meglumine antimoniate treatment on kidney function in dogs with clinical leishmaniosis.

This study examines correlations among serum proteins, clinical score, body weight and kidney function biomarkers after a standard treatment course (meglumine antimoniate plus allopurinol) in twelve Canine leishmaniosis (CanL) patients at the three times points pre treatment, after treatment and after the end of treatment. The laboratory variables measured were those used for the follow-up of sick dogs along with biomarkers of kidney function: glomerular filtration rate (GFR), creatinine (Cr), urea, calcium, inorganic phosphorus, urine specific gravity (USG) and urine protein to creatinine ratio (UPC). Arterial blood pressure (systolic blood pressure, SBP), clinical score (CS) and weight were also monitored over the study period. At Tp0, GFR was within the normal range in most dogs. Hyperfiltration was detected in three patients and hypofiltration in one. In dogs showing hyperfiltration, this factor remained in the non-azotemic range over the whole study period. After treatment normal filtration values were recovered. Meglumine antimoniate did not modify GFR or USG. A significant reduction in UPC was recorded. In all dogs, clinical scores improved. Negative correlation was found between GFR and Cr, UPC and albumin (Alb) and CS and Alb, while positive correlation was detected between UPC and total globulins (GlobT), CS and GlobT, UPC and total solids (TS), SBP and CS and SBP and UPC. Our findings indicate no impacts on kidney function of the treatment of CanL with meglumine antimoniate, as no effects were produced on GFR or USG. Treatment was effective and found to reduce UPC which could suggest improved glomerular injury.

Factors related to limitation of life support within 48h of intensive care unit admission: A multicenter study / Factores relacionados con la limitación del tratamiento de soporte vital en las primeras 48h de ingreso en unidades de cuidados intensivos: estudio multicéntrico

Objective: To determine factors related to limitations on life support within 48h of intensive care unit (ICU) admission. Study design: Prospective multicenter study. Setting: Eleven ICUs. Patients: All patients who died and/or had limitations on life support after ICU admission during a four-month period. Variables: Patient characteristics, hospital characteristics, characteristics of limitations on life support. Time-to-first-limitation was classified as early (<48h of admission) or late (≥48h). We performed univariate, multivariate analyses and CHAID (chi-square automatic interaction detection) analysis of variables associated with limitation of life support within 48h of ICU admission. Results: 3335 patients were admitted; 326 (9.8%) had limitations on life support. A total of 344 patients died; 247 (71.8%) had limitations on life support (range among centers, 58.6%-84.2%). The median (p25-p75) time from admission to initial limitation was 2 (0-7) days. CHAID analysis found that the modified Rankin score was the variable most closely related with early limitations. Among patients with Rankin >2, early limitations were implemented in 71.7% (OR=2.5; 95% CI: 1.5-4.4) and lung disease was the variable most strongly associated with early limitations (OR=12.29; 95% CI: 1.63-255.91). Among patients with Rankin ≤2, 48.8% had early limitations; patients admitted after emergency surgery had the highest rate of early limitations (66.7%; OR=2.4; 95% CI: 1.1-5.5). Conclusion: Limitations on life support are common, but the practice varies. Quality of life has the greatest impact on decisions to limit life support within 48h of admission

Objetivo: Determinar los factores relacionados con la limitación del tratamiento de soporte vital (LTSV) en las primeras 48h de ingreso en Unidades de Cuidados Intensivos (UCI). Diseño: Multicéntrico prospectivo. Ámbito: Once UCI. Pacientes: Pacientes fallecidos y/o en los que se aplicó LTSV durante 4 meses. Variables de interés: Características de pacientes, hospital y LTSV. Se definió LTSV precoz la que ocurría en las primeras 48h de ingreso y tardía >48h. Realizamos análisis univariado, multivariado y árbol de decisión chi-square automatic interaction detection (CHAID) con las variables asociadas con LTSV en las primeras 48h. Resultados: Incluidos 3.335 pacientes, en 326 (9,8%) se aplicó LTSV y 344 fallecieron; de estos 247 (71,8%) se limitaron (variabilidad interhospitalaria: 58,6-84,2%). La mediana de tiempo (p25-p75) entre el ingreso y la LTSV inicial fue de 2 (0-7) días. El análisis CHAID evidenció que la escala de Rankin modificada fue la variable más estrechamente relacionada con la limitación precoz. Entre los pacientes con Rankin>2 la LTSV precoz se realizó en el 71,7% (OR=2,5; IC 95%: 1,5-4,4) y la enfermedad pulmonar fue la variable más relacionada con la LTSV precoz (OR=12,29; IC 95%: 1,63-255,91). Entre los pacientes con Rankin≤2, la LTSV precoz ocurrió en el 48,8% siendo los pacientes con cirugía urgente aquellos con mayor LTSV precoz (66,7%; OR=2,4; IC 95%: 1,1-5,5). Conclusión: La LTSV es común pero la práctica es variable. La calidad de vida es la variable que mayor impacto tiene sobre la LTSV en las primeras 48h del ingreso en la UCI

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).

Factors related to limitation of life support within 48h of intensive care unit admission: A multicenter study.

OBJECTIVE: To determine factors related to limitations on life support within 48h of intensive care unit (ICU) admission. STUDY DESIGN: Prospective multicenter study. SETTING: Eleven ICUs. PATIENTS: All patients who died and/or had limitations on life support after ICU admission during a four-month period. VARIABLES: Patient characteristics, hospital characteristics, characteristics of limitations on life support. Time-to-first-limitation was classified as early (<48h of admission) or late (≥48h). We performed univariate, multivariate analyses and CHAID (chi-square automatic interaction detection) analysis of variables associated with limitation of life support within 48h of ICU admission. RESULTS: 3335 patients were admitted; 326 (9.8%) had limitations on life support. A total of 344 patients died; 247 (71.8%) had limitations on life support (range among centers, 58.6%-84.2%). The median (p25-p75) time from admission to initial limitation was 2 (0-7) days. CHAID analysis found that the modified Rankin score was the variable most closely related with early limitations. Among patients with Rankin >2, early limitations were implemented in 71.7% (OR=2.5; 95% CI: 1.5-4.4) and lung disease was the variable most strongly associated with early limitations (OR=12.29; 95% CI: 1.63-255.91). Among patients with Rankin ≤2, 48.8% had early limitations; patients admitted after emergency surgery had the highest rate of early limitations (66.7%; OR=2.4; 95% CI: 1.1-5.5). CONCLUSION: Limitations on life support are common, but the practice varies. Quality of life has the greatest impact on decisions to limit life support within 48h of admission.

Controlling phlebotomine sand flies to prevent canine Leishmania infantum infection: A case of knowing your enemy.

Leishmaniosis caused by Leishmania infantum is a widespread zoonotic disease that can be transmitted to animals and humans by their vectors, blood-sucking phlebotomine sand flies. To prevent canine leishmaniosis across the whole Mediterranean region, vector control is essential. Because of phlebotomine breeding sites are diverse, environmental larval controls have limited practical value. Control methods of adults are being evaluated, such as selective baits based on sugar feeding of males and females or Attractive Toxic Sugar Baits (ATSB), and the indoor use of Long-Lasting-Insecticidal Nets (LLINs) treated with permethrin to prevent sand fly bites complementing the Indoor Residual Spraying (IRS) approach suggested by WHO. Although several strategies exist, the best control measure to prevent canine Leishmania infantum is to treat dogs using biocidal topical formulations based on legal insecticides (PTs18) or repellents (PTs19) (as collars, spot-ons and/or sprays) during the period when the vectors are active. This means we need to really know the biology and life cycle of the sand fly vector. According to available data, by mapping ambient temperatures we can already predict high risk areas where vector densities will be higher. In ongoing research, new candidates are emerging to fight against sand flies including natural plant extracts with low impacts on the environment and host animal. Other options in the future could be systemic insecticides to help reduce sand fly populations in high density areas. In parallel, health authorities and professionals involved in animal and public health (veterinarians, physicians, entomologists and epidemiologists) must work together in a One Health approach to minimize Leishmania infection. Veterinarians play a crucial role in liaising between key stake holders and dog owners to ensure the latter act responsibly in using repellents as a preventive measure against sand fly bites.

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.

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.

Efficacy of 65% permethrin applied to dogs as a spot-on against Phlebotomus perniciosus.

Leishmania infantum is a protozoan parasite causing leishmaniosis, a visceral disease transmitted by the bites of sand flies. As the main reservoir of the parasite, dogs are the principal targets of control measures against this disease, which affects both humans and dogs. Several studies have revealed the usefulness of topical insecticide treatment (collars, spot-ons and sprays) in reducing the incidence and prevalence of L. infantum. The present study was designed to test the efficacy of 65% permethrin applied to dogs as a spot-on against the sand fly vector Phlebotomus perniciosus. The duration of the desired effects was also estimated to help design an optimal treatment regimen. Twelve dogs assigned to treatment (n=6) and control (n=6) groups were exposed to sand flies once a week over a seven-week period. Repellent and insecticidal efficacies were estimated and compared amongst the groups. Our findings indicate satisfactory repellent, or anti-feeding, effects lasting 3 weeks and short-term insecticidal effects lasting 2 weeks after initial application. Accordingly, we recommend the use of this product every 2-3 weeks during the active phlebotomine sand fly period to protect dogs against the bites of P. perniciosus.

Detection of zoonotic intestinal parasites in public parks of Spain. Potential epidemiological role of microsporidia.

Several studies have demonstrated that the soil of public parks presents an important source of infection which has a significant impact on public health. Children are the main group affected by accidentally ingestion of contaminated soil. This study was performed in order to identify the presence of zoonotic parasites in dog and cat faecal and soil samples from public parks of Madrid, Spain. Six hundred twenty-five and seventy-nine soil and faecal samples (presumably from dogs and cats) respectively were collected from 67 parks. Intestinal parasites were identified in 27 parks (40.3%), which were contamined with Giardia sp. (19.4%), microsporidia (19.4%), Toxocara spp. (16.4%), Cryptosporidium sp. (6%), Entamoeba histolytica (3%) and Ancylostomidae (3%). Combinations of two or more intestinal parasites were found in 11 parks, and it was common to find Giardia and microsporidia together in samples. Intestinal parasites were detected in 18% (112/625) of soil samples. The most frequent parasite species found in the examined soil samples were Toxocara spp. (16.4%), followed by Giardia sp. (4.5%) and Strongyloides sp. larvae (3%). The zoonotic parasites found in the 79 faecal samples were Giardia sp. (17.7%), Cryptosporidium sp. (9%), E. histolytica (2.5%), Trichuris vulpis (1.3%), Toxascaris leonina (1.3%) and microsporidia spores (28%). Microsporidia characterization by amplification of DNA confirmed 10 samples as positive, eight for E. bieneusi and two for E. hellem by PCR. The role of those parasites in the environment are discussed.

Questionnaire-based survey on the clinical management of canine leishmaniosis in the Madrid region (central Spain).

This paper describes a questionnaire designed to gain information on how veterinarians clinically manage canine leishmaniosis (CanL) in the Madrid region (central Spain). The present survey is one of the several similar questionnaire-based surveys conducted within the framework of the project EDEN (Emerging Diseases in a changing European eNvironment). The questionnaire sought to obtain data regarding the main clinical manifestations observed, the diagnostic methods used and the preventive measures recommended. Its Spanish version was sent by post to veterinary practitioners within the study area in two lots, one sent out in December 2006 and the other in March 2007. Only 174 of the 760 questionnaires sent were completed and returned (reply rate of 23%). Among the completed questionnaires, clinics differed widely in terms of features such as the habitats of the dogs (urban, peri-urban or rural) and patient volumes. Clinics attending dogs from peri-urban/rural habitats reported more suspected (p<0.001), confirmed (p=0.001) and newly diagnosed (p=0.001) cases/year than clinics providing service to a city clientele alone. According to the veterinary practitioners, skin lesions, lymphadenomegaly and weight loss were commonly observed, although these signs are not specific to CanL. Signs described to be of high diagnostic value were epistaxis and kidney disease. All the veterinarians polled reported that a suspicion of Leishmania infantum infection was confirmed by at least a serological method; the immunofluorescence antibody test (IFAT) being the technique most used. To prevent the disease, most vets recommended topical synthetic pyrethroids applied as impregnated collars or spot-ons. It is observed that despite considerable progress is being made in clinical management and controlling the disease, in Madrid Region its incidence continues to increase.

Seasonal trends and spatial relations between environmental/meteorological factors and leishmaniosis sand fly vector abundances in Central Spain.

This paper reports on an entomological survey performed over the period 2006-2008 in Central Spain (mainly in the Madrid province) where canine leishmaniosis (CanL) is endemic. The study area was selected on the grounds of its wide altitude range, which determines both broad climate and vegetation ranges that could affect sand fly distributions. This area was surveyed from NE to SW across its mountain range (Sistema Central) and plateau area using sticky traps mainly on embankments. In 2006 and 2007, 123 sites were sampled (9557 sand flies captured) to establish possible relations between environmental or meteorological factors and vector densities (Phlebotomus perniciosus and Phlebotomus ariasi). The factors correlated with higher vector densities were: a sample site between villages or at the edge of a village, the lack of a paved road, a rural habitat, an east or south-facing wall or wall sheltered from the wind, the presence of livestock or birds, a holm-oak wood vegetation, a lower summer mean temperature and lower annual mean precipitation. This study was followed by a seasonal survey conducted at 16 selected sites (14,353 sand flies) sampled them monthly from May to November 2008. P. perniciosus showed a diphasic seasonal trend with two abundance peaks in July and September whereas P. ariasi showed a monophasic trend with one peak in August. Comparing with data from studies performed in 1991 in the same area, vector densities are significantly higher. A possible explanation for this is that the vectors (mainly P. ariasi) are moving towards higher altitudes perhaps because of global change. This increasing trend could have an impact on CanL and its geographical distribution.

Emerging trends in the seroprevalence of canine leishmaniasis in the Madrid region (central Spain).

This report describes a cross-sectional serological survey of the epidemiology of canine leishmaniasis (CanL) performed in 2006 and 2007 in the Madrid region (central Spain) where the disease is endemic. The work presented here is one of the several studies conducted in different Spanish regions under the Integrated Project of the European Commission entitled Emerging Diseases in a changing European eNvironment (EDEN). The aim of this project is to identify and catalogue European ecosystems and environmental conditions that determine the spatial and temporal distributions and dynamics of several pathogenic agents including Leishmania infantum (EDEN-LEI). The study area (Madrid Autonomous Region) was selected on the grounds of its wide altitude range. This area was surveyed from NE to SW across its mountain range (Sistema Central) and plateau area. One thousand and seventy-six dogs from 32 villages were examined for clinical signs of CanL, and serum samples were obtained to determine several haematological and biochemical variables. Leishmaniasis-specific antibodies were identified using an indirect immunofluorescence antibody test (IFAT). 87 of the 1076 dogs were seropositive for the protozoan (IFAT: cut-off>or=1/80) indicating a seroprevalence of 8.1% (0-16.1% depending on the village). On the basis of a physical examination and the biochemical/haematological status of each dog, 32 of the 87 infected dogs were described as clinically healthy (37%). Seroprevalence showed a peak in young dogs (1-2 years) and a second larger peak among the older dogs (7-8 years). Factors correlated with a higher infection risk were age (OR=1.15 [95% CI: 1.07-1.22]), weight (OR=1.10 [95% CI: 1.04-1.16]), and living outdoors as opposed to in a home (OR=3.38 [95% CI: 1.42-8.05]). According to data from studies performed in 1992 in the same area, the seroprevalence of CanL has increased 1.54-fold [95% CI: 1.04-2.29]. Given that this increasing trend cannot be attributed to differences in the sociodemographic characteristics of the dog populations, it is proposed that environmental changes could have had an impact on vector and reservoir densities and their geographical distributions. Further studies designed to explain this trend should attempt to correlate sand fly densities and CanL seroprevalences with climate, land use and human changes.

Directions for the diagnosis, clinical staging, treatment and prevention of canine leishmaniosis.

Canine leishmaniosis (CanL) due to Leishmania infantum is a life threatening zoonotic disease with a wide distribution in four continents and importance also in non-endemic regions. The purpose of this report is to present a consensus of opinions on the diagnosis, treatment, prognosis and prevention of CanL in order to standardize the management of this infection. CanL is a disease in which infection does not equal clinical illness due to the high prevalence of subclinical infection among endemic canine populations. The most useful diagnostic approaches include serology by quantitative techniques and PCR. High antibody levels are associated with severe parasitism and disease and are diagnostic of clinical leishmaniosis. However, the presence of lower antibody levels is not necessarily indicative of disease and further work-up is necessary to confirm CanL by other diagnostic methods such as cytology, histopathology and PCR. We propose a system of four clinical stages, based on clinical signs, clinicopathological abnormalities and serological status. Suitable therapy and expected prognosis are presented for each of the stages. The combination of meglumine antimoniate and allopurinol constitutes the first line pharmaceutical protocol. However, although most dogs recover clinically after therapy, complete elimination of the parasite is usually not achieved and infected dogs may eventually relapse. Follow-up of treated dogs with blood counts, serum biochemistry, urinalysis, serology and PCR is essential for prevention of relapses. Protection against sand fly bites by topical insecticides is effective in reducing infection, and recent development of vaccines has indicated that prevention by vaccination is feasible.

Molecular characterization of Toxoplasma gondii isolates from cats in Spain.

Cats are important in the epidemiology of Toxoplasma gondii because felids are the only definitive hosts that can excrete environmentally resistant oocysts. Fresh samples of brain from 103 Spanish cats with antibodies to T. gondii were analyzed for T. gondii DNA using nested-PCR; 47 (45.5%) were found to be positive. Further characterization of DNA from 46 cats using RFLP-PCR at the 3' and 5' ends of the SAG2 locus revealed that 12 (26%) isolates were Type I and 34 (74%) were Type II; no Type III were found, and the 47th sample could not be classified to its genetic type. In addition, T. gondii was also isolated by bioassay in mice from 42 of 103 seropositive cats. This is the first report of T. gondii characterization from cats in Spain.

Seroprevalence and risk factors associated with Neospora caninum infection in different dog populations in Spain.

In this study, Neospora caninum seroprevalence and some associated risk factors were investigated in four different dog populations in Spain. N. caninum seropositivity was significantly higher in farm dogs (51%, 51/100) (P<0.001) and lower in household dogs (2.9%, 3/102) (P<0.0001). The seroprevalence in hunting (23%, 23/100) and stray (24.5%, 23/94) dogs was moderate, and no significant differences were observed between these two populations (P>0.05). A significantly higher number of dogs showed titres of 1:50-1:100 (68%, 68/100) than >or=1:200 (33%, 33/100) titres (P<0.0001). N. caninum antibodies were more often detected in mixed breed than pure breed dogs (P<0.01), but when data were stratified by dog type a significant association was not found (P>0.05). A significantly higher prevalence of N. caninum was observed in dogs over 1 year old (P<0.01), indicating that horizontal transmission may be the most important route of infection. The presence of N. caninum antibodies was significantly more frequent in Leishmania infantum-seropositive hunting (P<0.05) and stray dogs (P<0.00001). This study confirms that farm, stray and hunting dogs can be considered at-risk dog populations for N. caninum infection in Spain.

[Tetanus treated with continuous baclofen intrathecal perfusion]. / Tétanos tratado con perfusión continua de baclofeno intratecal.

After making a bibliographic search in our country and demonstrating the lack of original articles or communications on treatment with intrathecal baclofen for the generalized forms of tetanus, our objective is to present this treatment based on the successful experience of one clinical case. In relationship to the case, we review baclofen pharmacokinetic and pharmacodynamic effects and its possible utility in the treatment of tetanus.

Plasma electrophoretogram in feline immunodeficiency virus (FIV) and/or feline leukaemia virus (FeLV) infections.

The electrophoretogram of 89 cats, including those infected by feline immunodeficiency virus (FIV+), feline leukaemia virus (FeLV+) and non-infected, showed statistically significant differences in several of the fractions. FIV+ cats had very high protein values (mean, 8.10 g/dl), mostly because of hypergammaglobulinemia (mean, 2.81 g/dl) as compared with non-infected animals and FeLV+. In addition, in these FIV+ animals, the albumin/globulins ratio (A/G) was very low (mean, 0.72). Statistically significant differences in A/G and alpha2-globulin fraction were observed in FeLV+ group (A/G mean, 0.88 +/- 0.08; alpha2-globulin, mean, 0.84 +/- 0.07 g/dl) when compared with non-infected group (A/G mean, 1.06 +/- 0.08; alpha2-globulin mean, 0.68 +/- 0.04 g/dl). The alpha1-globulin fraction was higher in double infected animals (FIV and FeLV positive, F-F) (3.55 g/dl), than in FeLV+ or FIV+ cats (3.10 and 3.07 g/dl respectively), but no statistical conclusions may be drawn from this fact because of the low number of F-F animals. This technique may help to assess the initial clinical status of retrovirus-infected cats, and the clinical course of these chronic diseases, specifically during and after suitable therapy.

Tétanos tratado con perfusión continua de baclofeno intratecal / Tetanus treated with continuous baclofen intrathecal perfusion

Tras realizar una búsqueda bibliográfica en nuestro país y constatar la ausencia de comunicaciones o artículos originales acerca del tratamiento con baclofeno intratecal para las formas generalizadas de tétanos, nuestro objetivo es dar a conocer dicho tratamiento, a través de la experiencia de un único caso clínico, con buen resultado. En relación con el caso, revisamos la farmacocinética y farmacodinamia del baclofeno y su posible utilidad en el tratamiento del tétanos

After making a bibliographic search in our country and demonstrating the lack of original articles or communications on treatment with intrathecal baclofen for the generalized forms of tetanus, our objective is to present this treatment based on the successful experience of one clinical case. In relationship to the case, we review baclofen pharmacokinetic and pharmacodynamic effects and its possible utility in the treatment of tetanus