Identification of Sarcocystis spp. in Slaughtered Sheep from Spain and Evaluation of Bradyzoite Viability after Freezing.

Sarcocystis spp. are complex apicomplexan parasites that cause a substantial economic impact on livestock used for meat production. These parasites are present worldwide. Our study aimed to identify Sarcocystis species affecting sheep meat in southern-central Spain and to evaluate the effectiveness of freezing for parasite inactivation. A total of 210 condemned samples of sheep meat were thoroughly assessed grossly and microscopically; the presence of macro- and microcysts was confirmed. The samples were then frozen at -20 °C for various time intervals (24, 48, 72, 96, 120, and 144 h) and compared with untreated samples. Bradyzoites were isolated through pepsin digestion for subsequent molecular analysis and viability assessment, employing trypan blue and double fluorescence staining techniques. Our measurements confirmed the presence of S. tenella, S. gigantea, and S. medusiformis in Spanish domestic sheep. Freezing for 96 to 144 h resulted in a significant reduction in parasite viability, with a robust correlation observed between the two staining methods. Both stains effectively measured the viability of Sarcocystis, thereby promising future advances in meat safety.

Toxoplasma gondii in meat of adult sheep in Spain.

Toxoplasmosis is a zoonotic disease caused by Toxoplasma gondii, an intracellular parasite that presents a worldwide risk. Humans can become infected by ingesting meat infected with T. gondii, and the consumption of infected sheep and goat meat is a significant public health issue. Antibodies against T. gondii have been found in sheep in Spain, indicating the presence of the parasite in the country. However, no previous studies have assessed the presence of T. gondii in sheep meat in Spain. In view of the significance of the transmission of T. gondii through meat consumption and given the lack of previous studies in Spain, we carried out an investigation to evaluate the presence of T. gondii in adult sheep meat (mutton). A total of 216 muscle samples were analyzed by digestion, and a real-time PCR assay was used to determine the presence of T. gondii DNA. A total of 24.5% of the samples were found to be parasitized, indicating that the consumption of sheep meat can present an important risk for human health.

Preferences of Pet Owners in Regard to the Use of Insecticides for Flea Control.

A survey conducted from 2002 to 2012 of 759 customers from 84 veterinary practices allows us to examine the habits of dog and cat owners regarding the use of insecticides for flea control. The results indicate that the percentage of animals treated during the 12 mo prior to the survey was not very high (71% in dogs and 50% in cats), considering that 100% of animals included in the study were flea-infested. Statistical analysis shows that animals older than 4 mo are treated more frequently and that dogs are more likely to be treated than cats. Among cats, those living in houses with gardens or farms are treated at a higher rate than those that live in houses without gardens or in flats. The main variable associated with the degree of satisfaction and product loyalty is the active ingredient, and the choice of a commercial product is generally associated with high-efficacy products. The mode of application of a flea treatment does not seem to have a strong influence on pet owner satisfaction or loyalty. Likewise, we do not find a clear correlation between owners' satisfaction and loyalty, so pet owners may find the results of the selected product more or less satisfactory for reasons other than simple effectiveness. Finally, because of the complexity of the life cycle of these ectoparasites, the importance of the advice of a veterinarian when planning flea control is emphasized.

Toxoplasma gondii in raw and dry-cured ham: The influence of the curing process.

The aim of this work was to analyze Toxoplasma gondii in raw hams by mouse bioassay and to evaluate the effect of curing on the viability of the parasite to assess the risk of infection from eating dry-cured ham. After a serology study of 1200 pigs in Aragón (Spain), forty-one naturally infected pigs with different serological titers against T. gondii were selected. Two cured periods (9 and 12 months) were evaluated as well as the influence of the physicochemical composition of hams on T. gondii survival. Although the parasite burden was low, a high number of seropositive pigs with Toxoplasma tissues cysts in raw hams were found (31.6%). Viability of T. gondii was influenced by the curing, with statistically significant differences between fresh and cured hams (p < 0.001). The viability was higher in hams cured for 9 months compared to those cured for 12 months. However, this period of curing resulted in the reduction but not in a complete elimination of the risk. Thus, from a public health point of view, under the conditions of this study it is safer to consume dry-cured ham with periods of curing higher than 12 months. Analysis of physicochemical results did not identify any variable with significant influence on the presence and viability of T. gondii in cured ham, but loss of viability of T. gondii was observed in hams with a lower fat content. Further research is required to validate combinations of salts concentration and time of curing that can be used as preventive measures in the HACCP system of dry-cured ham industry.

Field efficacy of acaricides against Varroa destructor.

Field trials were conducted in Northeast Spain (Aragón) to evaluate the effectiveness of two acaricides against Varroa destructor. These experiments took into account the season of the year, apiary, colony, and developmental state and strength of the colony. The acaricides used were a synthetic (amitraz, Apivar®) and a natural (formulated from Api Life Var®, thymol oil and thymol alcohol) product. The treatments used in the present study reduce high infestations of V. destructor, although they do not eliminate the infestation. Similar efficacies between treatments were found. Nevertheless, the efficacy of a treatment depends on the apiary where applied. Moreover, the detected variability in the apiary and hive poses a challenge to the identification of the significant factors. Therefore, more field studies to assess efficacies in several apiaries are needed to obtain a better understanding of the effects of the applied treatments.

Toxoplasma gondii: Pig seroprevalence, associated risk factors and viability in fresh pork meat.

This study was conducted on 161 fattening pig farms located in Aragón (Northeast Spain). Serum samples from 1200 pigs were tested for antibodies against T. gondii by indirect immunofluorescence assay (IFA). Antibodies to T. gondii (≥1:20) were detected in 301 pigs (24.52%). The seroprevalence observed in the present study indicates a widespread exposure to T. gondii, as seropositive pigs were found in 96.67% of the farms studied although low pig titers were determined. Risk factors associated with T. gondii seroprevalence were presence of cats in or around the farms, presence of dogs around the facilities, low number of animals in the farms, poor hygiene and bad maintenance of the farms. Finally, it was observed that where rodent baits were used, Toxoplasma prevalence was lower. Risk management measures including control of cats and rodents on the farms, among others, could help to reduce the observed prevalence levels. By mouse bioassay, T. gondii was detected in 73.7% and isolated from 42.1% of seropositive pigs and a significant relation between the titers of pigs and the presence and viability of T. gondii in the tissues was found. The detection of T. gondii is not possible by currently practiced meat inspection. Nevertheless, the increased probability of detecting viable forms of T. gondii in tissues of pigs with titers ≥1: 80 could be used as the cutoff for discriminating higher risk animals, and could be used as an effective control tool for the industry of cured meat products. In practical terms, we propose that this value could be used as a critical limit in the HACCP system.

Prevalence of Rickettsia and Bartonella species in Spanish cats and their fleas.

The aim of this study was to determine the prevalence of Bartonella henselae, Rickettsia felis, and Rickettsia typhi in fleas and companion cats (serum and claws) and to assess their presence as a function of host, host habitat, and level of parasitism. Eighty-nine serum and claw samples and 90 flea pools were collected. Cat sera were assayed by IFA for Bartonella henselae and Rickettssia species IgG antibodies. Conventional PCRs were performed on DNA extracted from nails and fleas collected from cats. A large portion (55.8%) of the feline population sampled was exposed to at least one of the three tested vector-borne pathogens. Seroreactivity to B. henselae was found in 50% of the feline studied population, and to R. felis in 16.3%. R. typhi antibodies were not found in any cat. No Bartonella sp. DNA was amplified from the claws. Flea samples from 41 cats (46%) showed molecular evidence for at least one pathogen; our study demonstrated a prevalence rate of 43.3 % of Rickettsia sp and 4.4% of Bartonella sp. in the studied flea population. None of the risk factors studied (cat's features, host habitat, and level of parasitation) was associated with either the serology or the PCR results for Bartonella sp. and Rickettsia sp.. Flea-associated infectious agents are common in cats and fleas and support the recommendation that stringent flea control should be maintained on cats.

Use of new technologies to notify possible contagion of sexually-transmitted infections among men / Uso de las nuevas tecnologías para la notificación del posible contagio de infecciones de transmisión sexual entre hombres

Background: Among men who have sex with men (MSM), the association between searching for sexual partners’ on the Internet and increased risk of sexually transmitted infections (STIs)/HIV infection, together with current low levels of partner notification (PN), justifies a study to explore the intention to use new communication technologies for PN in Spain. Methods: Two cross-sectional surveys were performed: the first was administered online to visitors to web pages where the survey was advertised; the second was administered on paper to patients attending an STI Unit and centres similar to Community-Based Voluntary Counselling and Testing centres. Results: The study population comprised 1578 Spanish residents (median age, 34 years [range: 18 to 74]); 84% lived in urban areas, and 69% reported searching for sexual partners on the Internet. Thirty-seven per cent would be willing to use a website for PN, 26% did not know if they would use one, and 37% would not want to use one. The main reasons for not intending to notify STI/HIV were 'shame or fear' (stable partner) and 'not knowing how to contact them' (casual partner). The preferred method of notification was face to face (73%) for both stable and casual partners, although using new technologies (Short Messaging System, e-mail, web page, phone applications) was widely accepted for notifying casual partners. Conclusions: Fighting stigma and promoting alternative methods of PN among MSM and health professionals through new technologies could increase the frequency of PN. This approach will improve early detection and reduce transmission in Spain (AU)

Antecedentes: Entre los hombres que tienen sexo con hombres, la asociación entre buscar parejas por internet y el incremento de riesgo para presentar una Infección de Transmisión Sexual (ITS), junto con la baja frecuencia de la práctica de la notificación a las parejas sexuales expuestas a un infectado justifican explorar la intención de uso de las nuevas tecnologías para facilitar dicha notificación en España. Métodos: Se han utilizado dos estudios transversales: el primero, administrado 'online' a los visitantes de páginas web donde se publicitaba la encuesta; el segundo, administrado en formato papel en una Unidad Especializada de Atención a las ITS y otros centros similares a los de soporte y realización de pruebas. Resultados: Participaron 1578 personas residentes en España, mediana de edad 34 años (Rango: 18- 74); 84% de áreas urbanas, 69% buscaron parejas sexuales en internet. Un treinta y siete por ciento utilizarían un página web si estuviera disponible, un 26% no sabían si lo harían y otro 37% no la utilizarían. Las razones principales para no tener la intención de notificar de ITS/VIH a una pareja estable fueron: 'miedo o vergüenza'; a una pareja ocasional: 'no saber cómo contactarla'. El método preferido para notificar fue el 'cara a cara o por teléfono' (73%), tanto en caso de pareja estable como ocasional. El uso de nuevas tecnologías (SMS, e-mail, web page, aplicaciones de teléfono) fue más aceptado en caso de pareja ocasional. Conclusiones: La lucha contra el estigma y la promoción del uso de nuevas tecnologías para la notificación a las parejas sexuales, entre los hombres que tienen sexo con hombres (HSH) y por parte de los profesionales de la salud, pueden aumentar la notificación, con los beneficios esperados de una más temprana detección de casos y una reducción de la transmisión en España (AU)

Use of new technologies to notify possible contagion of sexually-transmitted infections among men.

BACKGROUND: Among men who have sex with men (MSM), the association between searching for sexual partners' on the Internet and increased risk of sexually transmitted infections (STIs)/HIV infection, together with current low levels of partner notification (PN), justifies a study to explore the intention to use new communication technologies for PN in Spain. METHODS: Two cross-sectional surveys were performed: the first was administered online to visitors to web pages where the survey was advertised; the second was administered on paper to patients attending an STI Unit and centres similar to Community-Based Voluntary Counselling and Testing centres. RESULTS: The study population comprised 1578 Spanish residents (median age, 34 years [range: 18 to 74]); 84% lived in urban areas, and 69% reported searching for sexual partners on the Internet. Thirty-seven per cent would be willing to use a website for PN, 26% did not know if they would use one, and 37% would not want to use one. The main reasons for not intending to notify STI/HIV were "shame or fear" (stable partner) and "not knowing how to contact them" (casual partner). The preferred method of notification was face to face (73%) for both stable and casual partners, although using new technologies (Short Messaging System, e-mail, web page, phone applications) was widely accepted for notifying casual partners. CONCLUSIONS: Fighting stigma and promoting alternative methods of PN among MSM and health professionals through new technologies could increase the frequency of PN. This approach will improve early detection and reduce transmission in Spain.

Neisseria gonorrhoeae: resistencias antimicrobianas y estudio de la dinámica poblacional. Situación en 2011 en Barcelona / Neisseria gonorrhoeae: Antimicrobial resistance and study of population dynamics. Situation in Barcelona in 2011

INTRODUCCIÓN: Las altas tasas de resistencia de Neisseria gonorrhoeae (NG) a ciertos antibióticos, junto con la aparición de cepas con sensibilidad disminuida y resistencia a las cefalosporinas, hacen de la infección gonocócica un problema de salud pública. Los objetivos del estudio fueron: realizar el seguimiento de la sensibilidad antimicrobiana de NG obtenidas entre enero y agosto de 2011, y estudiar su dinámica poblacional. MÉTODOS: Se estudió la sensibilidad mediante el método disco-difusión y E-test. El genotipado se realizó mediante el NG-MAST. RESULTADOS: De un total de 100 cepas, el 59% presentaron sensibilidad intermedia a penicilina y el 9% resistencia. Según EUCAST, se detectaron 3 gonococos con sensibilidad disminuida a ceftriaxona, 10 a cefixima y uno con resistencia de alto nivel a ambos (CMI 1,5 μg/ml). La CMI50 y la CMI90 a cefixima fue de 0,016 y de 0,125 μg/ml, mientras que a ceftriaxona fue < 0,016 y 0,064 μg/ml. El 99% presentó resistencia a doxiciclina, el 53% a ciprofloxacino, el 3% a azitromicina y el 1% a espectinomicina. El ST más prevalente fue el ST1407, mayoritariamente asociado a resistencia o sensibilidad disminuida a cefalosporinas o macrólidos. CONCLUSIÓN: NG ha desarrollado tasas importantes de resistencia a distintos antibióticos. Se ha detectado una cepa con resistencia de alto nivel a las cefalosporinas de tercera generación y varias con sensibilidad disminuida, además de observarse un aumento de la CMI50 y de la CMI90 a estos antibióticos. La estructura poblacional de NG permanece estable y común al resto de Europa, aunque se han identificado 2 nuevos secuencio tipos (ST7226 y ST7227) con potencial para seleccionarse y adquirir altos niveles de resistencia a cefalosporinas

BACKGROUND: Due to the high rates of antimicrobial resistance to certain antibiotics, together with the emergence of Neisseria gonorrhoeae (NG) with reduced susceptibility and resistance to third-generation cephalosporins, gonococcal infection is becoming a public health problem. The objectives of the study were: To keep track of the antimicrobial susceptibility of NG strains obtained from January to August 2011. To study the population dynamics. METHODS: The antimicrobial susceptibility was studied by disk-diffusion and E-test. The genotyping was performed by NG-MAST method. RESULTS: Of a total of 100strains studied, 59% showed intermediate sensitivity to penicillin and 9% were resistant. According to EUCAST, we detected 3gonococci with reduced susceptibility to ceftriaxone, 10 to cefixime and one with high-level resistance to both antibiotics (MIC 1.5 μg/ml). MIC50 and MIC90 to cefixime were 0.016 and 0.125 μg/ml, respectively, whereas to ceftriaxone they were < 0.016 and 0.064 μg/ml, respectively. Almost all (99%) of the strains were resistant to doxycycline, 53% to ciprofloxacin, 3% to azithromycin, and 1% to spectinomycin. The most prevalent ST was ST1407, predominantly associated to resistance or reduced sensitivity to cephalosporins or macrolides. CONCLUSIONS: NG has developed significant rates of resistance to various antibiotics. One strain has been detected with high level resistance to third generation cephalosporins, and several strains with reduced susceptibility. An increase in MIC50 and MIC90 to these antibiotics has also been observed. NG population structure remains stable and common to the rest of Europe, although two new ST (ST7226 and ST7227) have been identified that could be selected and acquire high levels of resistance to cephalosporins

[Neisseria gonorrhoeae: antimicrobial resistance and study of population dynamics. Situation in Barcelona in 2011]. / Neisseria gonorrhoeae: resistencias antimicrobianas y estudio de la dinámica poblacional. Situación en 2011 en Barcelona.

BACKGROUND: Due to the high rates of antimicrobial resistance to certain antibiotics, together with the emergence of Neisseria gonorrhoeae (NG) with reduced susceptibility and resistance to third-generation cephalosporins, gonococcal infection is becoming a public health problem. The objectives of the study were: To keep track of the antimicrobial susceptibility of NG strains obtained from January to August 2011. To study the population dynamics. METHODS: The antimicrobial susceptibility was studied by disk-diffusion and E-test. The genotyping was performed by NG-MAST method. RESULTS: Of a total of 100strains studied, 59% showed intermediate sensitivity to penicillin and 9% were resistant. According to EUCAST, we detected 3gonococci with reduced susceptibility to ceftriaxone, 10 to cefixime and one with high-level resistance to both antibiotics (MIC 1.5µg/ml). MIC50 and MIC90 to cefixime were 0.016 and 0.125µg/ml, respectively, whereas to ceftriaxone they were <0.016 and 0.064µg/ml, respectively. Almost all (99%) of the strains were resistant to doxycycline, 53% to ciprofloxacin, 3% to azithromycin, and 1% to spectinomycin. The most prevalent ST was ST1407, predominantly associated to resistance or reduced sensitivity to cephalosporins or macrolides. CONCLUSIONS: NG has developed significant rates of resistance to various antibiotics. One strain has been detected with high level resistance to third generation cephalosporins, and several strains with reduced susceptibility. An increase in MIC50 and MIC90 to these antibiotics has also been observed. NG population structure remains stable and common to the rest of Europe, although two new ST (ST7226 and ST7227) have been identified that could be selected and acquire high levels of resistance to cephalosporins.

Epidemiology of Oestrus ovis infection of sheep in northeast Spain (mid-Ebro Valley).

Slaughterhouse surveys to determine the prevalence and intensity of larval Oestrus ovis Linnaeus (Diptera: Oestridae) in sheep, were conducted monthly for 1 year in northeastern Spain. The prevalence of O. ovis in 120 sheep in northeast Spain was 84.2%. The monthly prevalence ranged from 62.5% in January to 100% in June and October. Of the 3,831 larvae collected, 3,005 were first instar larvae (78.4%), 744 second instar larvae (19.4%), and 82 third instar larvae (2.1%). The mean larval burden per infected sheep was 37.9, throughout the year although infection was higher in October. The high number of first stage larvae in winter (especially October, November, and December) may be due to a period of slower larval maturation. Third-stage larvae were always present but sometimes in very low numbers compared to the total. According to the results, oestrosis appears to be a serious problem in the region.

Evaluation of a specific immunochemotherapy for the treatment of canine visceral leishmaniasis.

The efficacy of specific immunochemotherapy against Leishmania infantum infection in dog was studied. The effects on transmission of the disease, as well as the cellular and humoral immune response were examined. The treated animals showed a significant reduction in the infection rates that were detected in Phlebotomus perniciosus females fed on the dog. The humoral immune response, assayed with an indirect immunofluorescence antibody test (IFAT), did not show significant variations under the influence of the therapy. The characterisation of the peripheral blood mononuclear cells (PBMC) using flow cytometry indicated a significant increase in the proportion of T lymphocytes, especially of CD4/TcR(alpha)(beta)(+) and CD4/CD45RA(+) cells, without showing evidence for modifications in the other leukocyte subsets. Cellular lymphoproliferation studies indicated a lack of a specific response to soluble leishmanial antigen (SLA), but the non-specific lymphoproliferative capacity assayed with phytohemagglutinin (PHA) was maintained.