Memoria para optar al Título Profesional de Médico Veterinario / La criptosporidiosis es una enfermedad parasitaria del tracto digestivo producida por protozoos del género Cryptosporidium. Aunque las especies de este agente se describieron desde comienzos del siglo veinte, solo a fines de éste, se ha reconocido como un patógeno ampliamente distribuido en diferentes animales y como una amenaza para la salud pública. Infecciones por este protozoario se encuentran descritas en todos los continentes, incluida la Antártica. En los humanos y otras especies, es considerada una zoonosis re-emergente. El primer brote de criptosporidiosis transmitido a través del agua, fue descrito en 1984, desde ese año numerosos autores han evidenciado la presencia de este endoparásito en aguas de superficie y residuales no tratadas, sugiriendo su rol como agente etiológico de diarrea. La persistencia y diseminación de éste protozoo en el ambiente, se sustenta en tres hechos biológicos: (1) Sus ooquistes son inmediatamente infectantes; (2) resiste condiciones medio ambientales y (3) al medir 4,5-5,4 μm, resistir a la acción de desinfectantes convencionales y al no ser es afectado por los procesos físicos de tratamientos de aguas, puede sobrevivir en ella por varios meses. A pesar de que existen diversas metodologías descritas en el mundo, para detectar este parásito en muestras de este tipo, el presente estudio sería el primer intento de desarrollar un protocolo estandarizado, hasta la fecha en Chile. Por lo anterior, el principal objetivo de este estudio estuvo dirigido a estandarizar en Chile un protocolo que permita la recuperación, concentración y detección de ooquistes de Cryptosporidium de origen animal y humano en muestras de aguas, usando las tinciones de Ziehl-Neelsen y Aureamina. Para esto, ooquistes de C. parvum aislados de heces de terneros y humanos diarreicos, previamente estudiados mediante Crypto-Strip® (inmunocromatografia) y PCR especie-específica, fueron artificialmente inoculados en muestras de aguas en concentraciones decrecientes. Se comparó la sensibilidad analítica de ambas tinciones, obteniéndose la mínima concentración de ooquistes de Cryptosporidium que son requeridos para dar una muestra como positiva. Los resultados, expresados en ooquistes/mL, muestran que ambas tinciones tienen una sensibilidad analítica similar, ya que son capaces de detectar hasta la concentración de 1.953 ooquistes en 250 mL, es decir, 7,8 ooquistes /mL. A partir de esto, se puede concluir que el uso de una u otra tinción en la pesquisa de este parásito en muestras de aguas, no se debe basar en la sensibilidad de las mismas, sino en ventajas y desventajas metodológicas para su implementación / Finaciamiento: Proyecto FIV 12101401.9102.006

El documento digital no refiere asesor / Manifiesta que la criptosporidiosis es una enfermedad parasitaria de distribución cosmopolita que es causante de severos brotes de diarrea en los rumiantes domésticos neonatales. El presente estudio evaluó por primera vez en campo el papel del Cryptosporidium parvum como factor de riesgo en la presentación de diarrea neonatal empleando el diseño epidemiológico de Caso-Control en alpacas del departamento de Puno. El muestreo se realizó entre los meses de Febrero y Marzo del 2006. Se recolectaron muestras fecales (n=487) directamente del recto de crías de alpacas entre 1 a 15 días de edad procedentes de las localidades de Antacalla, La Raya, Quimsachata y Macusani. Los frotis fecales fijados previamente en metanol, fueron procesados según la técnica de tinción de Ziehl-Neelsen Modificado. Se encontró que de los animales diarreicos, el 39 % (130/336) resultaron positivos a la infección por C. parvum. Por otro lado, el 23 % (35/151) de las alpacas sin diarrea estaban infectadas. El análisis de regresión logística demostró que el parásito no representó un factor de riesgo para diarrea neonatal en los animales estudiados (OR: 1.5, IC 95%: 0.9-2.4). Otras variables fueron también evaluadas junto a C. parvum, determinándose que la localidad de La Raya fue estadísticamente significativa (P < 0.05), representando un factor de riesgo para la presentación de diarrea neonatal (OR: 2.5, IC 95%: 1.1-6.1). / Tesis

Manifiesta que la criptosporidiosis es una enfermedad producida por el protozoo Cryptosporidium parvum, que se caracteriza por producir diarrea en los animales neonatos y ocasionar pérdidas económicas en la industria pecuaria. Afecta a un gran número de animales domésticos y silvestres e inclusive al hombre. El presente estudio tuvo como objetivo evaluar si la presencia de Cryptosporidium parvum es un factor de riesgo de diarrea en alpacas neonatas menores de 15 días de edad provenientes de diversas unidades alpaqueras del Departamento de Cuzco. Esta tesis empleó el diseño epidemiológico de Caso-control para establecer si existe una relación causal. Se tomaron 248 y 231 muestras fecales de animales con diarrea y de animales aparentemente sanos (n=497) durante la temporada de parición de alpacas del 2006. Las muestras fueron preservadas en una solución de Bicromato de Potasio al 2%. Para detectar la presencia de C. parvum se utilizó la Técnica de Tinción de Ziehl Neelsen Modificado (ZNM). Los datos se analizaron empleando una regresión logística con el paquete estadístico STATA 8.0. La regresión logística ajustó variables potencialmente confundentes como edad, sexo, raza y lugar de origen de las alpacas neonatas muestreadas. Se encontró un Odds Ratio de 4.3; I.C.= 2.3 – 7.9. Este estudio demuestra que las alpacas en las que se detectó la presencia de C. parvum, tienen 4.2 veces mayor predisposición a sufrir diarreas en relación a las alpacas con diagnostico ZNM negativo. Así mismo, se determinó asociación estadística significativa entre los animales que presentan infección por el parásito y los que manifiestan cuadros de diarrea (23.4%; n=58), en comparación con el grupo aparentemente sanos (8.6%; n=20). / Tesis

La criptosporidiosis es una enfermedad ocasionada por varias especies del género Cryptosporidium, se caracteriza por producir diarrea, sobre todo en neonatos y, dependiendo de la especie involucrada, puede ser zoonótica, llegándose a considerar un problema de salud pública. Diversos estudios fueron realizados en el Perú para poder determinar la prevalencia del Criptosporidium sp. encontrándose los mayores casos en los lugares que contaban con la mayor cantidad de animales. / Cryptosporidiosis is a disease caused by different species of the Cryptosporidium genus it overall produces diarrhea in newborns and depending on the specie involved, these can be involved in a problem of public health. Many studies were performed in Peru to determine the prevalence of Cryptosporidium and they showed that a major number of cases were in an area with a higher number of animals. The aim of this study was to establish if presence of Cryptosporidium parvum in mothers is a risk factor for presentation of Cryptosporidium parvum in offsprings.

Die Kryptosporidiose des Kalbes stellt eine orale Infektion mit dem obligat pathogenen Erreger Cryptosporidium parvum dar. Sie verursacht hauptsächlich bei Jungtieren unterschiedlich schwere und zuweilen tödlich verlaufende Diarrhoe. Zur Analyse einer integrierten Bekämpfungsmaßnahme unter den Bedingungen einer natürlichen Infektionsexposition wurden 123 Kälber eines landwirtschaftlichen Nutzbetriebes in Thüringen untersucht. Die Tiere wurden in zwei Gruppen randomisiert nach ihrem Geburtstermin aufgeteilt. Der Gruppe H+ waren 62 Tiere zugehörig welche in den ersten sieben Lebenstagen Halofuginon in einer Dosis von 120 µg/kg KGW oral verabreicht bekamen und in Neopredisan® desinfizierten Buchten aufgestallt wurden. Gruppe H- stellte mit 61 Kälbern die unbehandelte Kontrolle dar, deren Buchten ausschließlich mittels Hochdruckreinigung gesäubert wurden. Alle Tiere wurden in den ersten vier Lebenswochen gewogen beginnend am ersten Lebenstag. In der 25. Lebenswoche fand eine zusätzliche Gewichtsbestimmung statt wobei die Tiere der H+ Gruppe im Mittel um 5,8 kg (p>0,05) schwerer waren als H- Tiere. Die Kälber beider Gruppen wurden in den ersten vier Lebenswochen jeden dritten Lebenstag hinsichtlich der Oozystenausscheidung, des Ernährungszustandes, des Trinkverhaltens, des Dehydratationsgrades und der Körpertemperatur untersucht. Zusätzlich wurde die Lunge auskultiert und die Konsistenz der Kotproben beurteilt. Durch die Halofuginonbehandlung und der zusätzlichen Desinfektion fand in der H+ Gruppe eine signifikante Senkung der Oozystenausscheidung bis zum 10. Lebenstag statt, jedoch waren die Tiere nach Absetzen der Behandlung für eine Infektion voll empfänglich. Das Auftreten von Diarrhoe zeigte eine positive Korrelation zur Oozystenanzahl untersuchter Kotproben. Die Anzahl an Tagen mit Durchfall korrelierte positiv und signifikant mit der Oozystenausscheidung. Für den Ernährungszustand, das Trinkverhalten, der Körpertemperatur sowie weitere klinischer Parameter bestand zu keiner Zeit eine signifikante Beziehung zur Kryptosporidiose. Damit erwies sich die integrierte Bekämpfung aus Halofuginonbehandlung und gezielter Desinfektion im für die Kälber kritischen Zeitraum der ersten zwei Lebenswochen als überaus effektiv. Allerdings muß bei derart geschützten Kälbern anschließend aufgrund einer offensichtlich nicht ausreichenden Immunisierung mit einer höheren Empfänglichkeit für C. parvum gerechnet werden.

Reproduction de : Thèse de doctorat : Parasitologie : Lille 2 : 2008. / Résumé en français et en anglais. Titre provenant de l'écran-titre. Bibliogr. f. 178-195.

Apicomplexan parasites are responsible for some of the most devastating human and veterinarian diseases and are parasites of great economic importance. Apicomplexa include Plasmodium, Toxoplasma and Babesia species. The pathogenic mechanisms developed by Apicomplexa parasites, in particular those that reside in a parasitophorous vacuole, involve considerable changes to the host cell, including the expression of variable surface proteins required for immune evasion. In Plasmodium falciparum infections, host cell remodeling is responsible for disease symptomology and severity in the human host. This work represents a multi-faceted study of antigenic secretion and the role of secreted antigens in pathogenesis. We study in detail the mechanisms of antigen secretion in Apicomplexa parasites. By use of comparative genomics, we find Plasmodium Export Element (PEXEL)-like motifs in a subset of Cryptosporidium and Babesia secreted proteins. However, in Babesia the motif functions as a spherical body targeting sequence, suggesting that secretory mechanisms in Apicomplexa are adapted to the parasite's intracellular lifestyle. To elucidate the relationship and function of exported antigens, we first focused on P. falciparum to determine gene co-expression modules. We found that in vivo, export modules are composed of constitutively or variably expressed genes, the latter group associated with patient clinical phenotypes. We then focused on a novel gene family called "phist" and show, using transcriptional expression profiling, its role in P. falciparum cytoadherence. In total, we demonstrate that antigen secretion is an evolutionary mechanism in Apicomplexa parasites and that variant expression of the genes encoding these antigens may allow parasites to adapt to environmental stresses.

Enteric pathogens in pool water can be unintentionally ingested during swimming, increasing the risk of Acute Gastrointestinal Illness. Swimmer activities and behaviors influence pool water ingestion rates, and can be quantified for use in risk assessment. Enteric infection risk estimates help identify data gaps, areas to focus resources, and research needs. Primary objectives of this study were to develop electronic, self-administered "exposure" and "pool operations" questionnaires; to gather swimmer behavior and activity data for use in risk assessment; and to estimate Cryptosporidium parvum infection risk in swimmers. Results were used to identify data gaps and future research needs relative to treated recreational water. To achieve these objectives, 126 swimmers were recruited at four pool sites in Tucson, Arizona, video-taped, and asked to complete a post-swim questionnaire. Forty-six of the 126 swimmers submitted a 24 hr post-swim urine sample for quantifying pool water ingestion. Head submersion frequency and duration and splashes to the face were observed and quantified in video analysis, and activities and behaviors were reported on the exposure questionnaire. Variable data were analyzed for associations with pool water ingestion estimated by urinalysis. Results indicate questionnaires can be self-administered electronically; the exposure questionnaire can be used to estimate ingestion magnitude in place of urinalysis; leisure swimming activities (diving, playing, splashing, wading, sitting) and frequency of face splashes are ingestion exposure factors; and that Cryptosporidium infection risk is greatest among leisure swimmers. Other activities observed and suspected of having associations with ingestion were short submersion durations (<1 sec), and spitting and spouting water. More research and resources focused on improving treated recreational water environments and reducing risks among swimmers are needed. Developing an indicator organism test representative of Cryptosporidium, a monitoring program for treated recreational water, education aimed at leisure swimmer, and routine engineering and administrative controls are recommended. Swimming is a unique activity that can be enjoyed by people of all ages and abilities. Controlling hazards in pool environments reduces Recreational Waterborne Illness risks associated with pool water ingestion and improves the health and safety of swimmers.

His world is full of organisms with no names. Brian Leander is working to discover and characterize the diversity of life on Earth.

This research aimed to address the knowledge gaps in the literature regarding the removal of waterborne pathogens (viruses and protozoa) by modified multistage slow sand filtration. In the current study, two pilot-scale multistage slow sand filtration systems were operated continuously for over two years. The pilot systems treated agricultural- and urban-impacted raw river water of variable quality with turbidity peaks over 300 NTU and seasonal cold temperatures <2??C. The first system (Pilot 1) consisted of two independent trains that included pre-ozonation, shallow-bed upflow gravel roughing filtration, and shallow-bed slow sand filtration. Pilot 1 was a pilot-scale version of an innovative, commercially available full-scale system. The second system (Pilot 2) included a full-depth upflow gravel roughing filter, a full-depth slow sand filter, and a second shallow-depth slow sand filter in series. The SSFs of both pilots were operated at high hydraulic loading rates (typically 0.4 m/h) at the upper limit of the literature recommended range (0.05 to 0.4 m/h). Both pilot systems provided excellent turbidity removal despite the high filtration rates. Effluent turbidity of all multistage SSF pilot systems were within the regulated effluent limits in Ontario for full-scale SSFs (below 1 NTU at least 95% of the time and never exceeded 3 NTU), despite raw water turbidity peaks over 100 NTU. The roughing filters contributed to approximately 60-80% of the full-train turbidity removal, compared to and 20-40% for the slow sand filters. On average, the second slow sand filter in pilot 2 provided almost no additional turbidity removal. The slow sand filter run lengths were short because of frequent high raw water turbidity, with about 50-80% of the runs in the range of 1-3 weeks. To prevent excessive SSF clogging and maintenance, filtration rates should be decreased during periods of high turbidity. Seven Cryptosporidium and Giardia challenge tests were conducted on the slow sand filters of both pilot systems at varying filtration rates (0.4 or 0.8 m/h), temperatures (2 to 25??C), and biological maturities (4 to 20 months). Removal of oocysts and cysts were good regardless of sand depth, hydraulic loading rate, and water temperature in the ranges tested. Average removals in the SSFs ranged from 2.6 to >4.4 logs for Cryptosporidium oocysts and ranged from >3.8 to >4.5 logs for Giardia cysts. This was consistent with findings in the literature, where oocyst and cyst removals of >4 logs have been reported. Cryptosporidium oocyst removals improved with increased biological maturity of the slow sand filters. At a water temperature of 2??C, average removal of oocysts and cysts were 3.9 and >4.5 logs, respectively, in a biologically mature SSF. Doubling the filtration rate from 0.4 to 0.8 m/h led to a marginal decrease in oocyst removals. Sand depths in the range tested (37-100 cm) had no major impact on oocyst and cyst removals, likely because they are removed primarily in the upper section of slow sand filter beds by straining. In general, good oocyst and cyst removals can be achieved using shallower slow sand filter bed depths and higher filtration rates than recommended in the literature. There are very few studies in the literature that quantify virus removal by slow sand filtration, especially at high filtration rates and shallow bed depths. There are no studies that report virus removal by slow sand filtration below 10??C. As such, 16 MS2 bacteriophage challenge tests were conducted at varying water temperatures (<2 to >20??C) and filtration rates (0.1 vs. 0.4 m/h) between February and June 2006 on biologically mature slow sand filters with varying bed depths (40 vs. 90 cm). Biologically mature roughing filters were also seeded with MS2. Average MS2 removals ranged from 0.2 to 2.2 logs in the SSFs and 0.1 to 0.2 logs in the RFs under all conditions tested. Virus removal by slow sand filtration was strongly dependant on hydraulic loading rate, sand depth, and water temperature. Virus removal was greater at a sand depth of 90 cm vs. 40 cm, at an HLR of 0.1 m/h vs. 0.4 m/h, and at warm (20-24??C) vs. cold (<2-10??C) water temperatures when sufficient warm water acclimation time was provided. Increased sand depth likely increased MS2 removal because of greater detention time for predation and greater contact opportunities for attachment to sand grains and biofilms. A lower HLR would also increase MS2 removal by increasing detention time, in addition to decreasing shear and promoting attachment to filter media and biofilms. Greater MS2 removal at warmer water temperatures was attributed to improved biological activity in the filters. Schmutzdecke scraping was found to have only a minor and short-term effect on MS2 removals. Virus removal can be optimized by providing deep SSF beds and operating at low filtration rates. Virus removal may be impaired in cold water, which could affect the viability of using SSF/MSF at northern climates if communities do not use disinfection or oxidation. As a stand-alone process, slow sand filtration (with or without roughing filtration) may not provide complete virus removal and should be combined with other treatment processes such as disinfection and oxidation to protect human health.