Orientador: Ana Maria Aparecida Guaraldo / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-18T14:53:24Z (GMT). No. of bitstreams: 1 Hofstatter_PauloGonzalez_M.pdf: 2633496 bytes, checksum: 19c74e9814c573f8ebf8f7645de76a37 (MD5) Previous issue date: 2011 / Resumo: Psittacidae são aves de estimação bem conhecidas e comuns em zoológicos, parques e criatórios particulares. Têm uma ampla distribuição mundial, principalmente em regiões tropicais. Apesar de sua popularidade, pouco se sabe a respeito de seus parasitas, principalmente coccídios. O filo Apicomplexa é um grupo de protozoários predominantemente parasíticos de imensa importância médica e veterinária, o qual apresenta afinidades com Dinozoa, Ciliophora e Heterokonta. Apesar da presença de Apicomplexa em psitacídeos ter sido relatada diversas vezes, poucas espécies de Eimeria (6) e Isospora (2) foram descritas até o momento. O objetivo deste estudo foi investigar, identificar e descrever os parasitas coccidianos encontrados em aves psitaciformes. Para isto, fezes de aves cativas originárias de zoológicos e criadores particulares foram coletadas entre 2009 e 2010 e submetidas a técnicas de flutuação a fim de recuperar oocistos de coccidios e ovos de nematódeos. 43 das 237 amostras estavam positivas (18,1%). Oocistos típicos de Eimeria foram observados em Amazona aestiva, Ara rubrogenis e Platycercus sp. e uma nova espécie de Eimeria foi descrita em A. aestiva. Nenhum oocisto de Isospora foi observado. Além disso, tivemos a oportunidade de observar rica variedade de ovos de nematódeos, possivelmente pertencendo a Capillaria plagiaticia, Ascaridia sp., Heterakis sp. e outro material não identificado. Observou-se também que espécimes de zoológicos foram significativamente mais parasitados do que aves oriundas de criatórios particulares. A prevalência de nematódeos (16,9%) foi muito maior que de protozoários (2,1%) / Abstract: Psittacidae are well-known companion birds and common as zoo specimens, in parks and breeding facilities. They have a wide distribution around the world, mainly in tropical regions. Despite their popularity, not much is known about the parasites associated to them, mainly coccidia. The phylum Apicomplexa is a predominantly parasitic protozoan group of huge medical and veterinary importance, which bears evolutionary affinities to Dinozoa, Ciliophora and Heterokonta. Although the presence of apicomplexan parasites in psittacine birds was reported several times, only a handful of Eimeria (6) and Isospora (2) species has been described until now. The aim of this study was to investigate, identify and describe the coccidian parasites found in psittacine birds. For this, feces of captive birds originating from zoos and private breeders were collected from 2009 to 2010 and they were submitted to flotation techniques in order to recover coccidian oocysts and nematode eggs. 43 of 237 samples were positive (18,1%). Typical eimerian oocysts were observed in Amazona aestiva, Ara rubrogenis and Platycercus sp. and a new Eimeria species was described in A. aestiva. No isosporan oocysts were detected. Besides, we were able to observe a rich variety of nematode eggs, possibly belonging to Capillaria plagiaticia, Ascaridia sp., Heterakis sp. and another non-identified material. Zoo specimens were significantly more heavily parasitized than birds from private breeding facilities. Nematode prevalence was 16,9%, which was much more prevalent than that of coccidia, at about 2,1% / Mestrado / Parasitologia / Mestre em Parasitologia

No description available.

El presente estudio, tuvo por objetivo determinar el escore de lesiones intestinales macroscópicas de coccidias en pollos de engorde desafiados con cepas de campo locales de E. acervulina, E. maxima y E. tenella y suplementados con un programa anticoccidial (Salinomicina / Nicarbazina). El estudio se realizó en el galpón experimental del Laboratorio de Patología Aviar de la Facultad de Medicina Veterinaria de la Universidad Nacional Mayor de San Marcos; donde se utilizaron 600 pollos de engorde de la línea Cobb Vantress 500. El diseño comprendió 4 tratamientos: T1 (n=150), T2 (n=150), T3 (n=150) y T4 (n=150) con 6 repeticiones por tratamiento. T3 y T4 recibieron diferentes dosis del programa Salinomicina / Nicarbazina, 40 o 50 ppm de cada principio activo respectivamente, y fueron desafiados a los 14 días de edad, vía oral, con 1 mL de un inóculo con ooquistes de Eimerias colectadas de campo. Se registró mortalidad, signos clínicos y lesiones intestinales compatibles con coccidias desde los 14 a 28 días de edad. Al 7° día post desafío se determinó el escore de lesiones macroscópicas para coccidias utilizando la escala de + 0 a + 4 descrita en 1970 por Johnson y Reid. Se confirmó la presencia de especies de Eimeria mediante el raspado de la mucosa intestinal. En pollos de engorde, frente a un desafío con especies patógenas locales de Eimerias, el tratamiento anticoccidial demostró una significativa reducción de la mortalidad y signos clínicos por coccidiosis, y una significativa reducción del escore de lesiones intestinales macroscópicas de Eimeria acervulina y Eimeria maxima (p<0.05), mas no de Eimeria tenella. El análisis estadístico se realizó con el programa Stata 12.0 (Stata Corp). Palabras clave: Salinomicina, Nicarbazina, Eimeria sp., escore de lesiones, pollos de engorde / --- This study aimed to determine the score of macroscopic intestinal lesions of coccidiosis in broiler chickens challenged with strains of local field E. acervulina, E. maxima and E. tenella and supplemented with an anticoccidial program (Salinomycin / nicarbazin). The study was conducted in the experimental shed Avian Pathology Laboratory, Faculty of Veterinary Medicine of the National University of San Marcos; T1 (n = 150), T2 (n = 150), T3 (n = 150) and T4 (n = 150) with 6 replicates: 600 broilers where the Cobb Vantress line 500. The design included 4 treatments were used per treatment. T3 and T4 received different doses of Salinomycin / nicarbazin, 40 or 50 ppm of active ingredient each program respectively, and were challenged at 14 days of age, orally with 1 mL of an inoculum with oocysts collected Eimerias field. mortality, clinical signs and intestinal lesions compatible with coccidia was recorded from 14 to 28 days old. On the 7th day post challenge the score for coccidial lesions macroscopic scale using + 0 to + 4 described in 1970 by Johnson and Reid was determined. The presence of Eimeria species was confirmed by scraping the intestinal mucosa. In broilers, against a challenge with pathogenic Eimerias premises, the anticoccidial treatment demonstrated a significant reduction in mortality and clinical signs coccidiosis, and a significant reduction of the score of macroscopic intestinal lesions of Eimeria acervulina and Eimeria maxima (p <0.05), but not of Eimeria tenella. Statistical analysis was performed using Stata 12.0 (Stata Corp)program. Key words: Nicarbazin, Salinomycin, Eimeria sp., injury score, broilers

Estima la prevalencia de eimerias en cabras criollas de cuatro distritos del departamento de Ica. Además, determinar la frecuencia según las variables procedencia, estrato etario y sexo; estimar la asociación entre la carga parasitaria y las variables en mención e identificar las especies de eimerias presentes. Se colectaron 728 muestras de heces, entre los meses de julio y agosto del 2017, las cuales fueron procesadas en el Laboratorio de Microbiología y Parasitología-Sección Parasitología de la Facultad de Medicina Veterinaria de la Universidad Nacional Mayor de San Marcos; empleándose las técnicas coproparasitologícas cualitativa: flotación con solución de Sheather y cuantitativa: método de McMaster modificado. La identificación de las especies de eimerias se realizó mediante la medición de ooquistes esporulados, obtenidos por la técnica de Ritchie modificado. Se obtuvo una alta prevalencia de eimerias en la población evaluada de 99.2%. No se halló diferencias significativas entre la prevalencia y las variables edad, sexo y procedencia. El promedio de la carga parasitaria fue de 2 158 opg, considerada como carga baja. Se halló diferencia significativa entre carga parasitaria y las variables edad (<1 año) sexo (macho) y procedencia (El Carmen y Chincha Baja). Se identificaron 8 especies de eimerias en cabras, E. caprina, E. caprovina, E. ninakohlyakimovae, E. arloingi, E. apsheronica, E. jolchijevi, E. alijevi, y E. christenseni. / Tesis

An antibody library against Eimeria tenella sporozoites was constructed by phage display. Total RNA was isolated from the spleen, bone marrow, and ceca of immune chickens, and was used to reverse-transcribe cDNA. Heavy and light antibody variable genes were amplified from cDNA by the Polymerase Chain Reaction (PCR), using primer pairs that contain complementary sequences encoding a short linker sequence. The single-chain antibody fragment (scFv) was obtained by a secondary overlap PCR with primers that incorporate SfiI restriction sites, thus allowing for subsequent cloning into the phagemid vector pComb3X. Vector and scFv insert were digested with SfiI, ligated, and transformed into competent XL1-Blue Escherichia coli cells by electroporation, yielding a library with 7.4 x 107 total transformants. The culture was grown under carbenicillin selective pressure, rescued with helper phage, and the antibody-displaying phage was precipitated by PEG/NaCl, and subsequently used for panning. Five panning rounds were performed using cryopreserved E. tenella sporozoites, with a gradual increase of washing stringency to select for specific, highaffinity binders. A 1000-fold increase in phage output was obtained after 3 rounds of panning. There was clear enrichment of the positive clones over the panning rounds, with the 3rd round resulting in a 3,000-fold enrichment over the first one, as the binding clones became the dominant population in the library. Selected antibodies from the last round of panning were sequenced and characterized by immunoblotting. Soluble antibody fragments were produced in a non-suppressor E. coli strain, and recognized a 66-KDa sporozoite antigen on a Western blot. Primary cultures of chicken enterocytes were prepared in the hope of serving for invasion assays with E. tenella sporozoites. The isolation procedure, however, proved to be cumbersome and time-consuming. Future investigations will focus on purification and further characterization of antibodies selected from the constructed library. Such antibodies can be tested, alone or in combination, for their ability to block in vitro the invasion mechanism of E. tenella.

Two consecutive experiments were conducted to evaluate the influence of dietary composition, specifically protein and amino acid profile, on performance parameters, oocyst output, and lesion development in male and female replacement broiler breeders of two different genetic lines vaccinated with a live coccidiosis vaccine. Dietary formulations were based on either breeder specific recommendations or formulations of a broiler integrator. On day 28, males of each genetic line were added to female pens to evaluate the effect of co-mingling on male performance. Lesion assessment was performed on three separate occasions per each experiment. Fecal material was collected to determine oocyst cycling patterns. During experiment 1, flock uniformity was improved (P less than/equal to 0.05) in Line A males fed the integrator diet. Increased body weight and improved uniformity of Line B females was observed with the breeder recommended diet. Co-mingling negatively impacted (P less than/equal to 0.05) male body weight. Multiple oocyst peaks were observed in both genetic lines, with the first peak occurring at approximately 16 to 18 days post placement. This first peak tended to have the highest observed magnitude and corresponded with the highest level of intestinal lesions observed during the experiment. In experiment 2, diet impacted (P less than/equal to 0.05) average body weight in Line A males, Line B males, and Line B females. Line A males fed the breeder recommended diet had increased (P less than/equal to 0.05) body weight at the termination of the experiment. Line B males and females fed the breeder recommended diet had increased (P less than/equal to 0.05) body weights throughout the experiment beginning on day 7. Negative effects (P less than/equal to 0.05) on male body weight resulting from co-mingling were observed. Oocyst peaks were delayed and at a lower magnitude in both genetic lines compared to peaks observed in experiment 1. Dietary interactions were observed in both experiments where magnitude of peak, duration of oocyst output, and severity of lesion development was influenced by diet in both male and female genetic lines. These data indicate that co-mingling negatively impacts male performance and dietary composition can impact male and female performance, oocyst cycling, and lesion development during coccidiosis vaccination in replacement broiler breeders and should be taken into consideration when rearing replacement broiler breeders.

Abstract The purpose of this study is to investigate the probability of Eimeria tenella as a carrier for the transmission of chicken infectious anemia virus¡]CIAV¡^. One-day-old specific pathogen free¡]SPF¡^chickens were inoculated with CIAV, and then infected 20 days later with E. tenella. CIAV proteins were detected by immunofluorescent test in schizont, gametocytes and zygotes of E. tenella. Oocysts were purified from infected chicken by NaNO3 gradient centrifugation. Oocyst DNA was extracted by the treatments of SDS, proteinase K, DNase and chloroform. PCR template preparated from oocyst was used for amplification of CIAV DNA which is a 420 bp DNA fragment. The sequence of the PCR product from oocyst is similar to that of CIAV DNA from blood of infected chicken. The oocysts of E. tenella containing CIAV could also be suggested by inducing cytopathic effect after inoculated into cultured MSB1 cells. Our results indicated aprobability that E. tenella oocysts contain CIAV proteins and nucleic acids and thus E. tenella might paly an important role in the epizootiology.

Ziel der vorliegenden Arbeit war es, die Stallkokzidiose bei Sauglämmern in drei verschiedenen Lämmermastbetrieben anhand der klinischen Ausprägung und parasitologischer Parameter zu charakterisieren und zu vergleichen. Daneben wurden die Prävalenzen der am Infektionsgeschehen beteiligten Eimeria spp. untersucht und verglichen sowie morphometrische Merkmale ihrer Oozysten bestimmt. Des Weiteren wurde die Effektiviät einer metaphylaktischen Behandlung mit Toltrazuril im Vergleich zu unbehandelten Kontrolltieren untersucht. Ebenso wurden jeweils metaphylaktische und therapeutische Behandlungen mit Toltrazuril und Diclazuril miteinander und mit unbehandelten Kontrolltieren verglichen. Die Daten wurden im Zuge zweier Feldstudien erhoben, wobei die Kontrollgruppen für die epidemiologischen Untersuchungen genutzt wurden. In Studie 1, welche multizentrisch in drei verschiedenen Lämmermastbetrieben (A, B und C) in Sachsen-Anhalt und mit 120 Lämmern durchgeführt wurde, wurden zwei Gruppen gebildet: eine Kontrollgruppe und eine Toltrazuril-behandelte Gruppe, deren Lämmer metaphylaktisch eine einmalige orale Behandlung mit dem Wirkstoff (Baycox® 5% Suspension, Bayer HealthCare, Leverkusen) in der Dosierung von 20 mg/kg Körpergewicht etwa 12 Tage nach dem angenommenen Infektionszeitpunkt erhielten. Bei Studie 2, welche nur in Betrieb C stattfand, wurden aus weiteren 86 Lämmern drei Gruppen gebildet: eine metaphylaktische und eine therapeutische Diclazurilgruppe (Vecoxan®, Janssen, Belgien, Dosierung von 1 mg/kg Körpergewicht) und eine therapeutische Toltrazurilgruppe (Dosierung wie bei Studie 1). Diese wurden mit den Gruppen aus Studie 1 (Betrieb C) verglichen. Die therapeutische Behandlung wurde nach Beginn der Oozystenausscheidung (innerhalb von 48 Stunden nach Ausscheidung bei mindestens 10 % der Tiere der entsprechenden Gruppe) verabreicht. Es erfolgten über einen Zeitraum von 37 Tagen regelmäßige Entnahmen von Kotproben, bei denen die Oozystenausscheidung sowie die Kotkonsistenz bestimmt wurden. In Stichproben wurden die beteiligten Eimeria spp. differenziert. Ebenso wurden die Lämmer regelmäßig klinisch untersucht und die Gewichtsentwicklung anhand dreier Wägungen bestimmt. Kokzidiosen, welche sich subklinisch oder mit milden bis moderaten klinischen Symptomen manifestierten, traten in allen drei untersuchten Betrieben auf. Jedes Lamm schied mindestens einmal im Verlauf der Studie Oozysten aus, und die Erkrankung breitete sich rasch im Bestand aus. Ein typischer Verlauf der Befallsintensität, mit einem raschen Anstieg zu einem Maximum mit folgendem stetigen Absinken, konnte beobachtet werden. Insgesamt wurden 9 (Betrieb B) bzw. 12 (Betriebe A und C) Eimeria-Arten identifiziert. Es handelte sich (in absteigender Reihenfolge nach ihrer Prävalenz) um: E. ovinoidalis, E. faurei, E. parva, E. bakuensis, E. granulosa, E. crandallis, E. pallida, E. marsica, E. weybridgensis, E. ahsata, E. punctata und E. intricata. E. ovinoidalis war die prädominierende Art, sowohl bei der Höhe der Oozystenausscheidung und auch hinsichtlich der Prävalenz (> 90% in allen drei Betrieben). Das Auftreten von E. punctata in Deutschland wurde mit der vorliegenden Arbeit erstmals beschrieben. Infektionen mit drei bis fünf Eimeria spp. überwogen. Morphometrische Daten der Oozysten der nachgewiesenen Eimeria-Arten wurden dokumentiert und können als Hilfestellung für Diagnostik oder spätere Studien dienen. Durch die metaphylaktische Gabe von Toltrazuril wurden Höhe und Dauer der Oozystenausscheidung im Vergleich zur Kontrollgruppe in allen Betrieben signifikant reduziert (Studie 1). Im Hinblick auf die pathogenen Arten E. ovinoidalis und E. crandallis wurde eine signifikante Reduktion des Vorkommens sowie der Ausscheidungshöhe beobachtet. Ebenso wurde die Dauer des kokzidienassoziierten Durchfalls in der Toltrazuril-behandelten Gruppe signifikant im Vergleich zur Kontrollgruppe reduziert. In Studie 2 wurde die Oozystenausscheidung durch Toltrazuril und Diclazuril sowohl nach metaphylaktischer als auch therapeutischer Gabe signifikant reduziert. Die Ausscheidung von E. ovinoidalis-Oozysten wurde, mit Ausnahme der metapylaktischen Diclazurilgruppe, in den Behandlungsgruppen im Vergleich zur Kontrollgruppe gesenkt. Der kokzidienassoziierte Durchfall wurde ebenfalls in allen behandelten Gruppen im Vergleich zur Kontrollgruppe reduziert. Im Hinblick auf die untersuchten Parameter konnte für das Toltrazuril eine höhere Wirksamkeit im Vergleich zu Diclazuril festgestellt werden. Bezüglich der Gewichtsentwicklungen wurden in beiden Studien im Versuchszeitraum aufgrund der milden bis moderaten bzw. subklinischen Manifestation der Kokzidiose keine signifikanten Unterschiede festgestellt. Bei Lämmermastbetrieben mit intensiver Haltung ist eine Überwachung der Lämmer in der kritischen Phase (2-8 Wochen nach der Geburt) angezeigt, da mit Eimeria-Infektionen jedes Tieres und einer daraus resultierenden Kokzidiose zu rechnen ist und die Gefahr eines kritischen Vorkommens von E. ovinoidalis besteht. Für die medikamentelle Bekämpfung der Lämmerkokzidiose sind sowohl Diclazuril als auch Toltrazuril geeignet, wobei für das Toltrazuril eine deutlich höhere Reduktion der Oozystenausscheidung und des damit assoziierten Durchfalls festgestellt werden konnte. / The aim of the present studies was to characterise and compare coccidiosis in housed lambs of three different lamb fattening farms on the basis of the clinical symptoms and parasitological parameters. In addition, prevalences of the Eimeria spp. involved in the infection were determined and compared and morphological characteristics of their oocysts were determined. Furthermore, the efficacy of a metaphylactical treatment with toltrazuril was investigated in comparison to untreated control animals. Likewise efficacies of metaphylactical as well as therapeutical treatments with toltrazuril and diclazuril were compared with each other and to untreated controls. The data were collected in the course of two field studies, whereby the control groups were used for the epidemiological investigations. In study 1, which was conducted as a multicentric study in three different lamb fattening farms (A, B and C) in Saxony-Anhalt using 120 lambs, two study groups were formed: a control group and a toltrazuril-treated group. Lambs of the latter received a single oral treatment with toltrazuril (Baycox® 5% Suspension, Bayer HealthCare, Leverkusen) in a dose rate of 20 mg/kg body weight on the 12th day after the supposed infection time. In study 2, which was conducted only in farm C, three study groups with different treatment regimens were formed out of 86 additional lambs: a metaphylactical and a therapeutical diclazuril-group (Vecoxan®, Janssen, Belgien, dose rate 1 mg/kg body weight) and a therapeutical toltrazuril-group (dose rate as in study 1). These groups were compared with the study groups of study 1 (farm C). Therapeutical treatment took place after the onset of oocyst excretion (within 48 hours after 10% of the lambs of the respective groups showed oocyst excretion). During a time period of 37 days faecal samples were collected regularly and oocyst excretion and faecal consistency were determined. In randomly chosen samples the Eimeria spp. involved were differentiated. In addition, clinical examination of the lambs was carried out regularly and the body weight was determined at three time points of the study. Coccidiosis with subclinical or mild to moderate clinical manifestation occurred in all three investigated farms. Every lamb excreted oocysts at least once during the study, and the disease spread rapidly within the herd. A typical course of excretion intensity with a rapid incline to a maximum followed by a continuous decline, was encountered. A total of 9 (farm B) or 12 (farms A and C) Eimeria species were identified, respectively. The following species were recorded (in decreasing order according to their prevalence): E. ovinoidalis, E. faurei, E. parva, E. bakuensis, E. granulosa, E. crandallis, E. pallida, E. marsica, E. weybridgensis, E. ahsata, E. punctata and E. intricata. E. ovinoidalis was the predominant species both with regard to the oocyst excretion rate and the prevalence (> 90% in all farms). The occurrence of E. punctata in Germany was described by the present study for the first time. Infections with three to five Eimeria species were the most common. Morphometrical data of the oocysts were collected and may provide assistance for diagnostic purposes or prospective studies. In all farms rate and duration of oocyst excretion were significantly reduced by metaphylactical treatment with toltrazuril (study 1). With regard to the pathogenic species E. ovinoidalis and E. crandallis a significant reduction of occurrence and excretion rate was observed. Likewise, the duration of the coccidia-related diarrhoea in the toltrazuril-treated group was reduced significantly compared to the control group. In study 2 oocyst excretion was reduced significantly after both metaphylactical and therapeutical treatment with toltrazuril and diclazuril compared to the control group. Excretion of E. ovinoidalis-oocysts was reduced in the treated groups, except for the metaphylactical diclazuril group. Coccidia-related diarrhoea was reduced in all treated groups in comparison to the untreated control group. With respect to the examined parameters a higher efficacy of toltrazuril was observed compared to diclazuril. Regarding the development of body weight in both studies no significant group differences were observed due to the mild to moderate clinical symptoms of coccidiosis or due to the subclinical manifestation, respectively. In lamb-fattening farms with an intensive rearing system the monitoring of lambs in the critical phase (2-8 weeks after birth) is advisable because Eimeria-infection of every lamb resulting in coccidiosis have to be expected and a risk of critical occurrence of E. ovinoidalis has to be taken into account. For the control of lamb coccidiosis metaphylactical as well as therapeutical application of toltrazuril or diclazuril are suitable. However, toltrazuril showed a higher reduction of oocyst excretion and coccidia-related diarrhoea.

Zugl.: Giessen, Univ., Diss., 2006

Thesis (Ph. D.)--University of Wisconsin--Madison, 1956. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 51-53).