Loss of in vitro Efficacy of Topical Commercial Acaricides on Rhipicephalus microplus (Ixodida: Ixodidae) From Antioquian Farms, Colombia.

In Antioquia, the problems to control Rhipicephalus microplus (Canestrini, 1888) tick infestations have spread and ranchers claim conventional treatments are no longer effective. In this study, the in vitro efficacy of commercial topical products was tested with ticks obtained from two dairy farms in Antioquia with severe repeated infestations. About 800 engorged ticks were collected directly from animals in two separate visits at the beginning and end of the same month. The adult immersion test was used, which exposed groups of 40 ticks from each collection at the recommended concentration for five commercial products and combinations for 5 min. Efficacy was determined by comparing the reproductive index (fecundity × fertility) of each treated group to that of the control group. The values of all reproductive parameters obtained with ticks from the two collection dates were very similar. Cypermethrin (150 ppm) and amitraz (208 ppm) separately showed very low efficacies of only 10-20% at one farm, and zero at the other. The combination of chlorpyrifos + cypermethrin was the only product with an efficacy >50% at both farms and field observations corroborated to be still capable of eliminating infestations. Exposure to fluazuron at concentrations ranging from 0.05 to 500 ppm for 1 min reduced fertility in all groups by ≥99%, as would be expected for very susceptible strains. However, reduction of oviposition only occurred at the 500 ppm concentration. In conclusion, there is a high degree of resistance to all products tested except for fluazuron.

Effect of complementary Integrative Oncology on anxiety, depression and quality of life in thoracic cancer patients: A pilot study.

PURPOSE: Thoracic-cancer (TC) patients have an extensive toll taken in terms of health-related quality of life (HRQL). Integrative Oncology (IO) is recognized as a valid complement to any Standard-Therapy treatment to improve outcomes in TC patients. The objective of this pilot-study was to evaluate the effect of adding IO (acupuncture, Cognitive-Behavior Therapy and mindfulness) in terms of depression, anxiety and HRQL. METHODS: TC patients who attended the Thoracic Oncology-Unit from July to September 2018 were invited to participate. A total of 16 patients were included in the study, 8 patients were required to attend 5 weekly sessions of IO and 8 patients were invited as controls. Anxiety, depression and HRQL were measured at baseline and after completing 5 sessions of IO or 5 weeks, using the HADS scale and the QLQ-C30 scale. RESULTS: A total of 8 patients in the IO group attended the 5 sessions and 8 patients were followed as controls. Compliance with the therapies was high, none of the patients dropped out from the study. A tendency for improvement in anxiety, fatigue, pain and dyspnea was observed in patients attending IO, though not statistically significant likely due to sample size. CONCLUSIONS: IO therapies are well accepted among TC patients. Larger studies are necessary with robust sample sizes to improve conclusions regarding outcome improvements.

Multi-drug resistant Haemonchus contortus in a sheep flock in Antioquia, Colombia.

A sheep farm experiencing high lamb mortality and poor body conditions of ewes due to haemonchosis was tested for the presence of anthelmintic-resistant nematodes in August 2014. A fecal egg count reduction test was performed in Colombia on this farm and a DrenchRite® Larval development Assay (LDA) was shipped to the University of Georgia in the United States for resistance testing. The fecal egg count reduction test (FECRT) was conducted on 70 out of 500 animals allocated into five groups of 14 animals each: untreated control, albendazole (ABZ; 3.5mg/kg sc), ivermectin (IVM; 0.2mg/kg sc), moxidectin (MOX; 0.2mg/kg sc) and levamisole (LEV; 5mg/kg sc) plus triclabendazol/ivermectin (TBZ; 10mg/kg and IVM; 0.2mg/kg po). Individual fecal egg counts (FEC) using the modified McMaster technique and pooled larval cultures were performed 10days after anthelmintic treatment. Following 10days the initial results from the ABZ and IVM groups, 24 animals were treated twice with trichlorfon (TCF; 50mg/kg po) on days 0 and 6, and then checked for FEC on day 10. Mean differences in FEC before and after treatment were negligible (3 to 8% change) for the control, ABZ, LEV-TBZ and IVM groups. The MOX treatment resulted a mean FEC reduction of 76.7%, which was not-significant by t-test (p=0.08). The TCF treatment showed a mean FEC reduction of 42.3%, which was also not-significant (p=0.1). The DrenchRite® LDA was used to evaluate the resistance status to benzimidazole, levamisole, ivermectin and moxidectin on this farm. Coprocultures indicated that the predominant worm species was Haemonchus contortus (Colombia 94-100%) (UGA 77% H. contortus and 23% Trichostrongylus colubriformis). Resistance was evaluated for both H. contortus and T. colubriformis. The LDA results indicated high resistance to all 4 anthelmintics for both parasite species. The current study, together with similar surveys in 4 other Antioquian goat and sheep farms, confirms the presence of multi-drug resistant H. contortus to all three classes of anthelmintics in this region of Colombia.

Survey of gastrointestinal parasites, liver flukes and lungworm in feces from dairy cattle in the high tropics of Antioquia, Colombia.

A cross sectional study was undertaken to determine the prevalence and intensity of parasitic infections in dairy cattle in the high tropics of Colombia. A total of 1003 rectal samples were collected from dairy cows at 29 farms between May and June 2014 to represent the number of farms, age groups, and size of the 65,000-cow population in the municipality of San Pedro de los Milagros. Coprological techniques were used to detect gastrointestinal nematodes, liver flukes, coccidian oocysts, and first larval stage counts of Dictyocaulus viviparus. In order of decreasing prevalence, the following parasites were detected: coccidial oocyst (36.7%; 95% CIs, 31.6-42.7), strongyle nematodes (31.6%, 27.8-35.4), liver flukes (30.9%, 21.5-37.5), cestodes (8.4%, 7.1-9.7), and D. viviparus (5.4%, 3.4-7.5). Co-infections by all possible combinations of the three most predominant groups occurred in 11 to 15% of the animals. There were significant differences in infection rates between age groups, with higher risk of liver fluke infection in animals older than 1 year of age (odds ratio (OR) = 3.2), but lower presence for coccidia and strongyles (OR = 0.19 and 0.51, respectively). For Fasciola hepatica, within-herd prevalences of > 25% in 16 farms and 94 of 281 (33.5%) animals with > 5 eggs per gram (epg) indicate that significant production losses are likely occurring. The variation in the prevalence of gastrointestinal parasites and liver flukes, together with the level of infection among age groups, could be used in integrated management programs to establish selective anthelmintic treatments and select for heritable traits of host resistance. These results serve as a baseline for future studies to determine the success of control measures and should increase awareness that subclinical parasitism is widespread in the livestock sector.

Ubiquitination dynamics in the early-branching eukaryote Giardia intestinalis.

Ubiquitination is a highly dynamic and versatile posttranslational modification that regulates protein function, stability, and interactions. To investigate the roles of ubiquitination in a primitive eukaryotic lineage, we utilized the early-branching eukaryote Giardia intestinalis. Using a combination of biochemical, immunofluorescence-based, and proteomics approaches, we assessed the ubiquitination status during the process of differentiation in Giardia. We observed that different types of ubiquitin modifications present specific cellular and temporal distribution throughout the Giardia life cycle from trophozoites to cyst maturation. Ubiquitin signal was detected in the wall of mature cysts, and enzymes implicated in cyst wall biogenesis were identified as substrates for ubiquitination. Interestingly, inhibition of proteasome activity did not affect trophozoite replication and differentiation, while it caused a decrease in cyst viability, arguing for proteasome involvement in cyst wall maturation. Using a proteomics approach, we identified around 200 high-confidence ubiquitinated candidates that vary their ubiquitination status during differentiation. Our results indicate that ubiquitination is critical for several cellular processes in this primitive eukaryote.

Cross-sectional study of 13 Leptospira serovars in cows in a Colombian dairy region / Estudio transversal de 13 serovares de Leptospira en vacas en una región lechera colombiana / Estudo transversal de 13 sorotipos de Leptospira em vacas em uma região leitera colombiana

Abstract Background: Leptospirosis in cattle is recognized as a major cause of reproductive loses due to abortions, early embryonic death, and infertility. The type of Leptospira serovars that are currently circulating in Colombian herds is largely unknown due to the lack of national reports. Objective: The prevalence of antibodies specific to 13 Leptospira spp serovars was investigated in unvaccinated cows from 26 herds in San Pedro de los Milagros (Province of Antioquia) dairy region. Methods: Microscopic agglutination test was used on serum samples, following WHO recommendations. Results: A total of 112/900 (12.4%) of the animals had antibody titers of 1:100 or greater to at least one Leptospira serovar. Of those positive, 34/900 (3,8%) animals seroconverted to two or more serovars. In order of decreasing prevalence, the serovars detected were: hardjo prajitno, pomona, grippotyphosa, tarassovi, copenhageni, canicola, cellodeni and bratislava. There were 28 animals seropositive to serovar hardjo. Conclusion: Our results indicate that leptospirosis should be included as a differential diagnosis for various reproductive problems in this region.

Resumen Antecedentes: La leptospirosis es la mayor causal de pérdidas reproductivas debidas a abortos, muerte embrionaria temprana e infertilidad. Se desconocen los serovares de Leptospira que actualmente se encuentran circulando en el rebaño colombiano debido a la falta de reportes nacionales. Objetivo: Se investigó la prevalencia de anticuerpos específicos para 13 serovares de Leptospira spp en vacas no vacunadas de 26 rebaños en la región lechera de San Pedro de los Milagros (Provincia de Antioquia). Métodos: La prueba de aglutinación microscópica fue empleada en muestras de suero de acuerdo a las recomendaciones de la OMS. Resultados: Un total de 112/900 (12,4%) de los animales presentaron títulos de anticuerpos de 1:100 o superiores para al menos un serovar de Leptospira. Entre los positivos, el 34/900 (3,8%) de los animales presentaron seroconversión para dos o más serovares. En orden decreciente de prevalencia, los serovares detectados fueron: hardjo prajitno, pomona, grippotyphosa, tarassovi, copenhageni, canicola, cellodeni y Bratislava. En total, 28 animales fueron seropositivos al serovar hardjo. Conclusión: La leptospirosis debería ser incluida como diagnóstico diferencial en problemas reproductivos en esta región.

Resumo Antecedentes: Aleptospirose em bovinos é reconhecida como uma das principais causas de perdas reprodutivas devido a abortos, morte embrionária precoce e infertidade. Os sorotipos de Leptospira que estão atualmente em circulação no rebanho colombiano não se conhecem devido à falta de reportes nacionais. Objetivo: Neste estúdio foi investigada a prevalência de anticorpos específicos para 13 sorovares de Leptospira spp em vacas não vacinadas de 26 rebanhos leiteiros da região de San Pedro, Antioquia. Métodos: Foi empregado o teste de aglutinação microscópica (MAT) em amostras de soro de acordo com as recomendações da OMS. Resultados: Um total de 112/900 (12,4%) animais apresentaram títulos de anticorpos de 1:100 ou superiores para pelo menos um sorotipo de Leptospira. Dos animais positivos, 34/900 (3,8%) soroconverteram para dois ou mais sorotipos. A fim de diminuir a prevalência, foram detectados os sorotipos: hardjo, prajitno, pomona, grippotyphosa, tarassovi, copenhageni, canicola, cellodeni e Bratislava. Um total de28 animais foram soropositivos para o sorotipo hardjo. Conclusão: Os resultados indicam que a leptospirose deveria ser incluída nos diagnósticos diferenciais para o diagnóstico diferencial de problemas em doenças reprodutivas nos bovinos de Antioquia.

PREVALENCIA DE Babesia caballi, Theileria equi Y TRIPANOSOMIASIS Y ANÁLISIS DE FACTORES DE RIESGO EN EQUINOS DE ANTIOQUIA, COLOMBIA / PREVALENCE OF Babesia caballi, Theileria equi AND TRYPANOSOMIASIS AND ANALYSIS OF RISK FACTORS IN HORSES OF ANTIOQUIA, COLOMBIA

RESUMEN Esta investigación, se llevó a cabo, debido a la poca información nacional existente en hemoparásitos equinos y su objetivo fue determinar la prevalencia de babesiosis y tripanosomiasis, en el Valle de Aburrá y Rionegro, municipios de Antioquia (Colombia), así como algunos factores de riesgo asociados a la presentación de seropositividad, a estas enfermedades. En 223 predios, con una población de 1.008 equinos, se tomó muestra de sangre venosa, para realizar el diagnóstico serológico y molecular. La información epidemiológica fue obtenida, a través de la aplicación de una encuesta dirigida a las personas a cargo de los animales. Para el análisis estadístico, se aplicó Chi cuadrado de independencia y la Prueba Exacta de Fisher, cuando fue necesario. Para las asociaciones bivariadas, se calcularon estimaciones del riesgo (OR) por variable explicativa, con sus respectivos intervalos de confianza, del 95% (I.C. 95%). Se encontró una prevalencia del 11,9%, para babesiosis y de 1,9%, para tripanosomiasis; como factor de protección, se encontró el hecho de salir a una feria. Para la piroplasmosis, el lugar geográfico, el sexo, estar castrado, ser positivos a Anemia Infecciosa Equina (AIE), estrongílidos u oxiurus fueron factores de riesgo. Para tripanosomiasis, el factor de riesgo fue estar infestado con Dermacentor nitens o ser mular. Se deben reforzar métodos de vigilancia epidemiológica activa, sobre todo, en casos de movilización continua de equinos o desarrollo de eventos, que involucren presencia masiva de ejemplares.

ABSTRACT This research was carried out due to the little existing national information on equine hemoparasites; the objective was to determine babesiosis and trypanosomiasis prevalence in the Aburrá Valley and Rionegro Antioquia (Colombia) as well as some risk factors associated with the seropositivitie of these diseases. In 223 lots with a population of 1.008 equine venous blood sample was taken for serological diagnosis and molecular. Epidemiological information was obtained through the application of a survey addressed to persons in charge of the animals. For the statistical analysis, Chi square of independence or Fisher's Exact Test was applied when necessary. For the bivariate associations, risk estimates (OR) were calculated by explanatory variable, with their respective 95% confidence intervals (95% CI).It was found a prevalence of 11.9% for babesiosis and 1.9% for trypanosomiasis and as protection factor found the fact of going to trade fairs. Geographic location, sex and being neutered, be positive for Equine Infectious Anemia (AIE), strongyles or oxiurus was a risk factor for piroplasmosis. For Trypanosomiasis the risk factor was to be infested with ticks Dermacentor nitens or be mule. Methods of active epidemiological surveillance should be reinforced, especially in cases of continuous mobilization of equines or development of events that involve massive presence of horses.

The ubiquitin-activating enzyme (E1) of the early-branching eukaryote Giardia intestinalis shows unusual proteolytic modifications and play important roles during encystation.

Giardia intestinalis is considered an early-branching eukaryote and is therefore a valuable model for studying primordial cellular processes. This work reports the characterization of the ubiquitin-activating enzyme (E1) during growth and different stages of trophozoite differentiation into cysts. We found that in Giardia E1 expression (both at mRNA and protein levels) is regulated during encystation. The enzyme is proteolytically processed mainly into two fragments of 68kDa (N-terminal) and 47kDa (C-terminal). This phenomenon has not been described for any other E1. In trophozoites, this enzyme localized at spots within the cytoplasm as detected by using polyclonal antibodies against either E1 N- or C-terminal fragments. This pattern changed during encystation into a diffuse localization throughout the cytoplasm of encysting cells. E1 localizes in mature cysts at cytoplasmic spots and in the cyst wall. Our antisense silencing experiments suggested that E1 is an essential gene for parasite viability. On the other hand, E1 over-expression greatly increased the encystation rate, indicating a relationship between E1 and Giardia differentiation.

Ivermectin resistance of three Rhipicephalus microplus populations using the larval immersion test / Resistencia de tres poblaciones de Rhipicephalus microplus a la ivermectina por el ensayo de inmersión de larvas / Resistência de três populações de Rhipicephalus microplus à ivermectina pelo teste de imersão de larvas

Background: in Colombia, the control of tick infestation in cattle is almost exclusively performed with chemical acaricides. It is important to determine the degree of resistance of Riphicephalus microplus field populations to ivermectins (IVM) as the first step to design strategies to increase the useful life of acaricides and decrease the reliance on chemical controls. Objective: to test the degree of resistance to IVM. Methods: three cattle farms in Antioquia (Colombia) where IVM had previously failed to control infestations of R. microplus ticks were studied. Ticks were collected several months apart in 2013, and the larval immersion test (LIT) was performed on the progeny of the adult females. Concentration-mortality data were subjected to probit analysis. Results: the three populations showed lethal concentrations (LC) 50 and 99 of ≥ 30 ppm and ≥ 400 ppm, respectively. There was no difference observed in the LC at different times of collection for each population studied. Such high values for LC, together with very low regression slopes (≤ 2), indicated a very heterogeneous response to increasing concentrations of IVM, which is common for resistant populations. At two farms, subcutaneous injection with a long-acting formulation of IVM at a dosage of 630 μg/Kg was ineffective at eliminating existing infestations and protecting against reinfestations, confirming farmer suspicions of lost efficacy. Conclusion: epidemiological studies are necessary to assess the current status of resistance to IVM in this region of Colombia, and it is likely that the intensive use of IVM will aggravate this situation in the future.

Antecedentes: en Colombia, el control de la infestación de garrapatas en el ganado se ha realizado casi exclusivamente con acaricidas químicos y es importante determinar el grado de resistencia de las poblaciones de campo de Riphicephalus microplus a ivermectinas (IVM) como el primer paso para diseñar tácticas que aumenten su vida útil y disminuyan la dependencia de los controles químicos. Objetivo: probar el grado de resistencia a IVM. Métodos: se evaluó la resistencia de R. microplus a IVM en tres fincas ganaderas en Antioquia (Colombia) donde se sospechaba que productos con IVM estaban fracasando para controlar infestaciones por garrapatas. Se recolectaron garrapatas con varios meses de diferencia durante el año 2013 y se empleó la prueba de inmersión de larvas (LIT) con la descendencia de las hembras adultas. Los datos de concentraciónmortalidad fueron sometidos a análisis probit. Resultados: los resultados para las tres poblaciones estudiadas mostraron concentraciones letales (LC) 50 y 99 de ≥ 30 ppm y ≥ 400 ppm, respectivamente. No se observó diferencia en las LC obtenidas en diferentes momentos de recogida para cada población estudiada. Los altos valores de LC, junto con pendientes muy bajas en las rectas de regresión (≤ 2), indicaron una respuesta muy heterogénea a concentraciones crecientes de IVM, propio de poblaciones resistentes. En dos de las granjas, la administración de inyecciones subcutáneas con una formulación de acción prolongada de la IVM a dosis de 630 μg/Kg fue incapaz de eliminar las infestaciones y proteger frente a reinfestaciones, confirmando las sospechas de los ganaderos de pérdida de eficacia. Conclusión: estudios epidemiológicos son necesarios para evaluar el estado actual de la resistencia a IVM en esta región de Colombia, y es probable que el uso intensivo de IVM agrave esta situación en el futuro.

Antecedentes: na Colômbia, o controle de infestação com carrapatos em bovinos tem sido realizado quase exclusivamente com acaricidas químicos pelo qual se faz importante para determinar o grau de resistência de populações de campo de Rhipicephalus microplus a uma das drogas antiparasitárias de maior uso no manejo de gado, a ivermectina (IVM), como o primeiro passo para projetar táticas para aumentar a sua vida útil e diminuir a dependência de controles químicos. Objetivo: testar o grau de resistência à ivermectina. Método: foi avaliada a resistência de R. microplus à ivermectina em três fazendas de gado leiteiro localizadas no departamento de Antioquia (Colômbia) com suspeita de que o controle dos carrapatos com a utilização deste produto não estava sendo efetivo. Coletaram-se carrapatos em diferentes meses durante o ano 2013, e foi utilizado o teste de imersão de larvas (LIT) na descendência das fêmeas adultas, como método para detecção da resistência. Os dados de concentração-mortalidade foram submetidos à análise Probit. Resultados: todas as três populações avaliadas mostraram concentrações letais (LC) 50 de ≥ 30 ppm, e LC 99 de ≥ 400 ppm. Não foram observadas diferenças na LC entre as coletas feitas nos diferentes meses, em cada população amostrada. Os altos valores de LC encontrados junto com uma baixa regressão linear (≤ 2), indicam uma resposta muito heterogênea as concentrações crescentes de ivermectina, próprio de populações resistentes. Em duas das fazendas, a administração de injeções subcutâneas de ivermectina com uma formulação de longa ação, em doses de 630 μg/Kg, não foram capazes de eliminar as infestações e proteger contra a reinfestação, confirmando as reclamações dos fazendeiros para a perda de eficácia da droga antiparasitária. Conclusões: os estudos epidemiológicos são necessários para avaliar o estado atual da resistência à ivermectina nesta região da Colômbia, e é provável que o uso intensivo da ivermectina agrave ainda mais esta situação no futuro.