L’objectiu general d’aquesta tesi és l’estudi de les característiques reproductives del mascle i de la femella del burro Català per aplicar tecnologies reproductives que potenciïn la seva cria i evitin l’extinció d’aquesta raça de burro. El primer treball va estudiar el comportament sexual, les característiques del cicle estral i la predicció de l’ovulació en la burra Catalana. Els signes de zel més característics observats en les burres foren l’obertura i tancament de la boca junt amb l’extensió de les orelles enrere, orinar i vulvejar. Es va determinar que la durada del cicle estral era de 24.9 ± 0.3 dies, on el zel tenia una durada de 5.6 ± 0.2 dies. Les ovulacions es produïen amb un diàmetre mig preovulatori de 44.9 ± 0.5 mm i es va observar un percentatge elevat d’ovulacions múltiples (44.3%). La combinació dels següents paràmetres: diàmetre fol·licular, textura fol·licular i comportament de zel van determinar-se com els millors predictors de l’ovulació. El segon treball va estudiar l’efecte de la dilució i la centrifugació del semen (dos mètodes d’eliminació del plasma seminal) sobre la viabilitat, la motilitat i l’estructura subpoblacional mòtil del semen de burro Català refrigerat a 5ºC durant 24, 48 i 72h. Es va observar que la centrifugació o una ràtio de dilució elevada (1:10) eren els tractaments que mantenien més òptimament la viabilitat espermàtica durant les primeres 48h de refrigeració. L’estructura de 4 subpoblacions espermàtiques mòtils es mantenia amb la dilució o centrifugació, ara bé, només les mostres centrifugades després de 24h de refrigeració (5ºC) mantenien exactament la mateixa proporció d’espermatozoides en cada subpoblació que les mostres de semen fresc. El tercer treball va avaluar els canvis en la viabilitat i la motilitat que pateixen els espermatozoides de ruc Català durant la refrigeració (2h) i posterior centrifugació a tres temperatures diferents (5, 15 i 20ºC). En tots els tractaments es van mantenir les 4 subpoblacions mòtils del semen del ruc, observant que la refrigeració i centrifugació a 20ºC era la que mantenia l’estructura subpoblacional espermàtica més similar al semen fresc de burro Català. El quart estudi va valorar els efectes de la congelació-descongelació del semen de burro Català en comparació amb el semen de porc, sobre l’estructura subpoblacional mòtil i la viabilitat dels espermatozoides. Les característiques de motilitat dels espermatozoides de burro i de porc van respondre de manera molt diferent al procés de congelació-descongelació, mantenint però en els dos casos l’estructura de 4 subpoblacions mòtils. Els canvis en els valors mitjans de motilitat en les dues espècies estudiades es van deure sobretot, a canvis en les proporcions dels espermatozoides inclosos en cada subpoblació específica. En l’últim treball es va desenvolupar un model per avaluar la funcionalitat espermàtica del semen fresc i congelat de burro Català mitjançant l’interacció dels espermatozoides amb oòcits bovins lliures de zona pel·lúcida. Els espermatozoides de burro van ser capaços de fusionar-se amb l’oolemma i descondensar-se i formar el pronucli masculí (85-94%). Es va observar que les mostres de semen fresc i descongelat amb una viabilitat elevada (>60%) tenien índex de penetració superiors a les mostres amb viabilitat baixa (<40%). Així doncs, es van obtenir correlacions positives significatives (P<0.01) entre el percentatge de fecundació i la viabilitat (r=0.84), i també amb alguns paràmetres de motilitat (VAP, r=0.56; VCL, r=0.61; and mean ALH, r=0.68). En conclusió, una predicció adequada de l’ovulació de les burres, juntament amb l’utilització dels mètodes òptims de maneig i congelació del semen de ruc Català contribuiran positivament a l’aplicació de tecnologies reproductives en programes de cria que ajudin a evitar l’extinció d’aquesta raça. / The aim of this work is the knowledge of Catalonian donkey reproductive features of its females and males in order to apply reproductive technologies to maximize the production of live foals and help conserve the breed. In the first work we studied the reproductive behaviour, oestrous cycle characteristics and indicators that predict the moment of ovulation of the Catalonian donkey. The main signs of oestrus detected were mouth clapping, urinating and contraction and relaxation of the labia of the vulva with eversion of the clitoris. The length of the oestrus cycle was 24.9 ± 0.3 days, with oestrus itself lasting 5.6 ± 0.2 days. The mean diameter of the preovulatory follicle at day -1 was 44.9 ± 0.5 mm and we observed high incidence of multiple ovulations (44.3%). The combination of follicle size, follicle texture and oestrous behaviour were the best predictors of ovulation. In the second study we evaluated the effects of dilution and centrifugation (two methods of reducing the influence of the seminal plasma) on the survival of spermatozoa and the structure of motile sperm cell subpopulations in refrigerated Catalonian donkey semen at 24, 48 and 72h. At 48h, more surviving spermatozoa were seen in the more diluted (1:10) and in the centrifuged semen samples. The four donkey motile sperm subpopulations were maintained by refrigeration and centrifugation, moreover only centrifuged samples and only at 24h (5ºC) showed exactly the same motile sperm subpopulation proportions as recorded for fresh sperm. In the third work we assessed the effect of three storage temperatures (5, 15, 20ºC) on the survival and motility of Catalonian donkey sperm during transport (2h) to the laboratory for centrifugation. All treatments maintained the four motile sperm subpopulation, but 2h of storage at 20ºC followed by centrifugation was the treatment that maintained subpopulation structure more similar than fresh samples. In the fourth study we evaluated the influence of freeze/thawing on motile sperm subpopulations and sperm viability in ejaculates from two phylogenetically different mammalian species, boar and donkey. Boar and donkey sperm respond very differently in their mean motion characteristics to freezing/thawing, this process did not change the existence of a 4-subpopulations structure in the ejaculates. Motility changes induced by freezing/thawing protocol are especially linked to changes in the specific percentage of each of the motile sperm subpopulations. In the last work we studied the interaction between fresh/frozen-thawed donkey spermatozoa and zona pellucida (ZP)-free bovine oocytes in an attempt to develop a model for assessing cryopreserved Catalonian donkey sperm function. The donkey spermatozoa were able to fuse with the oolema and even to decondense and form the male pronucleus (85-94%). Fresh or frozen-thawed high viability (>60%) spermatozoa had higher penetration percentage than fresh or frozen/thawed low viability (<40%) spermatozoa. A significant positive correlation (P<0.01) was detected between percentage fertilization and viability (r=0.84), and between percentage fertilization and certain CASA parameters (VAP, r=0.56; VCL, r=0.61; and mean ALH, r=0.68). In conclusion, the exact moment prediction of ovulation in Catalonian jennies and adequate donkey semen manipulation and cryopreservation should have a positive impact upon the reproductive technologies application. Such knowledge should help maximize the production of live foals and help conserve the breed.

Wh-indefinites in Chinese have received great attention and discussion in the literature. This thesis investigates the status of Chinese wh-indefinites and their behaviors in the so-called donkey sentence. A typical example of wh-indefinite will be like the following: (4) Ta bu xiang zai shuo shenme le he not want again say what LE ¡¥He does not want to say anything again.¡¦ ¥L¤£·Q¦A»¡¤°»ò¤F¡C In this thesis, accounts of wh-indefinites as variables, polarity items and quantifiers from Huang (1982), Li (1992), Cheng (1991, 1994) and Lin (1996, 1998) are provided and this thesis discusses the problems each account presents. Countering Li¡¦s (1992) work, this work argue that wh-indefinite like zenmeyang ¡¦how¡¦ and weishenme ¡¥why¡¦ cannot be directly treated as variables since they feature variables only if they appear after an auxiliary (under which situation they will express purpose or method) in a donkey sentence according to Tsai (1999, 2000), see the following examples: (5) a. Akiu hui wei (le) shenme cizhi wo jiu hui wei (le) shenme cizhi¡@ Akiu will for LE what resign I then will for LE what resign ¡¥If Akiu will resign for the purpose x, I will then also resign for the purpose x.¡¦ ªüQ·|¬°¡]¤F¡^¤°»òÃ㾡A§Ú´N·|¬°¡]¤F¡^¤°»òÃ㾡C b. *Akiu wei (le) shenme hui cizhi wo jiu wei (le) shenme hui cizhi Akiu for LE what will resign I then will LE what will resign Intended ¡¥If Akiu will resign because of the reason x, I will then also resign because of the reason x. *ªüQ¬°¤F¤°»ò·|Ã㾡A§Ú´N¬°¤F¤°»ò·|Ã㾡C (Tsai¡¦s 2000, 15 glossed and translated by the author) Avoiding treating wh-indefinites as variables, we may treat them as quantifiers according to Huang¡¦s observation. But Huang¡¦s (1982) quantifier account of wh-indefinites is also rejected because I observe that wh-indefinites do not exhibit island sensitivity. Then we are led to another treatment: treating wh-indefinites as polarity items because they are sensitive to polarity environment. Aside from the sensitivity to polarity environment, Lin¡¦s (1996) work argues that wh-indefinites are licensed as polarity items if the sentence is subject to NEEC- non-entailment of existence condition. NEEC tells us that if (part of) a sentence does not have the existential import of object; wh-words can be licensed as wh-indefinites. This faces challenge because there are cases presupposing existential import yet the wh-indefinite is licensed. In chapter 3, two types of donkey sentences are identified in Cheng and Huang¡¦s (1996) work and are accounted with different approaches-Unselective Binding and E-type analysis. Though accepting their viewpoint generally, Lin (1996) makes a distinction between one-case and multi-case reading and opens the possibility of universal interpretation in ruguo-conditionals. Later I introduce Indirect binding approach to account both types of donkey sentences. Indirect Binding argues that some quantified expression plays the role as associating the indefinite with the anaphoric element in donkey sentences: (6) [Everyone who ti keeps a dogj]i like itj. The example here captures the very basic idea of Indirect Binding: the chunk of expression, which has the indefinite a dog in its scope, c-commands the pronoun it and hence relates it to the indefinite. I extend it to the analysis of Chinese donkey sentences and discover that although Indirect Binding does not target at a particular status, the condition that the indirect binder must c-command the indefinite seems to suggest that c-commanding is the way of licensing a wh-indefinite, which echoes Li¡¦s and Cheng¡¦s analyses. In Chapter 4, I introduce wh-indefinites in Japanese and Korean respectively and conclude that quantificational force comes from the environment but not from wh-indefinites themselves, countering Huang (1982). Finally, I suggest that licensing a wh-word as a wh-indefinite is through being c-commanded by the relevant operators. It is so because of Li¡¦s and Cheng¡¦s observation of wh-indefinites licensing and also of a condition in Indirect Binding that indirect binders must c-command the indefinite in order to have it as in its scope, as I have mentioned. This thesis then provides a viewpoint that perhaps, wh-indefinites can be treated uniformly as polarity items considering that Indirect Binding approach explains their behavior in donkey sentences. This thesis also opens the door for analyzing wh-indefinites in donkey sentences under Indirect Binding in other languages. Keywords¡G Wh-indefinite, polarity item, variable, quantifier, donkey phenomenon, Indirect Binding

Made available in DSpace on 2014-06-11T19:23:42Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-11-27Bitstream added on 2014-06-13T19:30:08Z : No. of bitstreams: 1 toledo_czp_me_jabo.pdf: 2114280 bytes, checksum: 3353189f0d92e6cea2a99b3bf82c63cc (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Objetivou-se com este estudo analisar a morfologia placentária de fêmeas asininas, espécie, cuja importância no meio rural é de inestimável valor, sendo poucas as informações detalhadas sobre sua placenta, “órgão” imprescindível à viabilidade fetal e a interação materno-fetal, mediante estudo macroscópico, microscópico e de microscopia eletrônica de varredura, os quais, exceto a macroscopia, foram realizados também na interação materno-fetal (útero-placentária). Para tanto, utilizaram-se onze fêmeas desta espécie e foram colhidas 11 placentas logo após a parição a campo, no instante de sua liberação, além de fragmentos de uma placenta oriunda de uma dessas 11 fêmeas, que teve sua gestação acompanhada por duas vezes. Assim, na primeira delas realizou-se operação cesariana e fragmentos da interação materno-fetal foram colhidos; parte deles fixados para posterior processamento usual em histologia e, outra parte, fixada e processada para análise à microscopia eletrônica de varredura. Sua segunda gestação foi acompanhada até o momento da parição e a liberação da placenta após parto normal, sendo colhida na íntegra. Após a colheita, as onze placentas foram pesadas, analisadas macroscopicamente e alguns fragmentos placentários fixados para subseqüente processamento usual em histologia; outros fragmentos foram fixados convenientemente para posterior processamento à microscopia eletrônica de varredura. Das peças analisadas e das preparações microscópicas e ultraestruturais, foram realizadas fotografias, fotomicrografias e ultrafotografias para a documentação. Verificou-se que o valor médio dos pesos dos jumentos, como os pesos das placentas, em relação aos neonatos foi de 8.36.1.10 e 13,18% respectivamente... / The objective of this study was analyzing the placental morphology of female asinines, species whose importance in the rural area is priceless, being little the detailed information about their placenta, vital “organ” to fetal viability and maternal fetal interaction, upon macroscopic, microscopic study, and scanning electron microscopy, which, except the macroscopy, were performed also in the maternal fetal interaction (placental-uterus). Therefore, eleven females of this species were used and eleven placentae were collected right after the calving down at field, in the instant of their release, besides the placenta fragments of one of these eleven females, which had its gestation followed-up twice. Thus, in the first of them, a caesarian operation was performed and fragments of the maternal fetal interaction were collected; part of them fixed for future usual processing in histology and, the other part, fixed and processed for analyses to scanning electron microscopy. Its second gestation was followed up until the moment of calving and the liberation of the placenta after the normal calving, being collected at full. After being collected, the eleven placentae were weighed, analyzed macroscopically and some placental fragments fixed for later usual processing in histology; other fragments were fixed conveniently for future processing to scanning electron microscopy. Among the analyzed pieces and the microscopic and ultra structural preparations, photographs, photomicrographs and ultra photographs were taken for the documentation. It was found that the average weights of the donkeys, as the weights of the placentae, in relation to the neonates were 8.36.1.10 and 13.18% respectively... (Coplete abstract click electronic access below)

Resumo: Objetivou-se com este estudo analisar a morfologia placentária de fêmeas asininas, espécie, cuja importância no meio rural é de inestimável valor, sendo poucas as informações detalhadas sobre sua placenta, "órgão" imprescindível à viabilidade fetal e a interação materno-fetal, mediante estudo macroscópico, microscópico e de microscopia eletrônica de varredura, os quais, exceto a macroscopia, foram realizados também na interação materno-fetal (útero-placentária). Para tanto, utilizaram-se onze fêmeas desta espécie e foram colhidas 11 placentas logo após a parição a campo, no instante de sua liberação, além de fragmentos de uma placenta oriunda de uma dessas 11 fêmeas, que teve sua gestação acompanhada por duas vezes. Assim, na primeira delas realizou-se operação cesariana e fragmentos da interação materno-fetal foram colhidos; parte deles fixados para posterior processamento usual em histologia e, outra parte, fixada e processada para análise à microscopia eletrônica de varredura. Sua segunda gestação foi acompanhada até o momento da parição e a liberação da placenta após parto normal, sendo colhida na íntegra. Após a colheita, as onze placentas foram pesadas, analisadas macroscopicamente e alguns fragmentos placentários fixados para subseqüente processamento usual em histologia; outros fragmentos foram fixados convenientemente para posterior processamento à microscopia eletrônica de varredura. Das peças analisadas e das preparações microscópicas e ultraestruturais, foram realizadas fotografias, fotomicrografias e ultrafotografias para a documentação. Verificou-se que o valor médio dos pesos dos jumentos, como os pesos das placentas, em relação aos neonatos foi de 8.36.1.10 e 13,18% respectivamente... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The objective of this study was analyzing the placental morphology of female asinines, species whose importance in the rural area is priceless, being little the detailed information about their placenta, vital "organ" to fetal viability and maternal fetal interaction, upon macroscopic, microscopic study, and scanning electron microscopy, which, except the macroscopy, were performed also in the maternal fetal interaction (placental-uterus). Therefore, eleven females of this species were used and eleven placentae were collected right after the calving down at field, in the instant of their release, besides the placenta fragments of one of these eleven females, which had its gestation followed-up twice. Thus, in the first of them, a caesarian operation was performed and fragments of the maternal fetal interaction were collected; part of them fixed for future usual processing in histology and, the other part, fixed and processed for analyses to scanning electron microscopy. Its second gestation was followed up until the moment of calving and the liberation of the placenta after the normal calving, being collected at full. After being collected, the eleven placentae were weighed, analyzed macroscopically and some placental fragments fixed for later usual processing in histology; other fragments were fixed conveniently for future processing to scanning electron microscopy. Among the analyzed pieces and the microscopic and ultra structural preparations, photographs, photomicrographs and ultra photographs were taken for the documentation. It was found that the average weights of the donkeys, as the weights of the placentae, in relation to the neonates were 8.36.1.10 and 13.18% respectively... (Coplete abstract click electronic access below) / Orientador: Gilson Hélio Toniollo / Coorientador: Márcia Rita Fernandes Machado / Banca: Marion Burkhardt de Koivisto / Banca: Marcos Lania de Araújo / Mestre

O objetivo do presente estudo foi avaliar a inclusão de duas fontes energéticas (alto açúcar e alto amido (AA), alta fibra e alto óleo (FO)) na dieta de asininos e equinos, através de ensaios de digestibilidade aparente dos nutrientes, características físico-químicas das fezes e parâmetros sanguíneos, vinculados ao metabolismo energético dos equídeos. Foram utilizados 20 equídeos, dez equinos da raça Quarto de Milha e dez asininos da raça Pêga, distribuídos no delineamento alternado do tipo Cross-Over, com duas fontes energéticas e duas espécies. As dietas experimentais foram compostas de 50% da energia proveniente do Feno Tifton 85 (Cynodon spp.) e 50% proveniente do concentrado, formulados a base amido e açúcar, e fibra e óleo. Foi observado efeito de dieta (p&lt;0,05) sobre os coeficientes de digestibilidade do extrato etéreo e fibra em detergente ácido, capacidade tamponante a pH 5 e pH 6 (maiores valores para dieta Fibra óleo) e sobre as concentrações sanguíneas de glicose (maiores valores para dietas açúcar amido. Observou-se efeito de espécie (p&lt;0,05) sobre o coeficiente de digestibilidade aparente de proteína bruta, sobre a capacidades tamponante à pH 5 e sobre as concentrações sanguíneas de glicose (maiores níveis glicêmicos observados para equinos). Houve efeito de tempo de coleta (p&lt;0,05) sobre as concentrações plasmáticas de acetato. Para as concentrações sanguíneas e fecais de ácidos graxos de cadeia curta totais, propionato não houve efeito (p&gt;0,05) de dieta ou de espécie. Dietas composta por predominância de volumoso acrescido de concentrado a base de amido e açúcar e fibra e óleo são igualmente aproveitadas pelos equídeos com destaque para a dieta fibra e óleo no aproveitamento de lipídios, sem causar efeito deletério sobre as características físico-químicas, o pH e a capacidade tamponante das fezes. Asininos são mais eficientes no tamponamento de ácidos intestinais. Os asininos apresentam maiores concentrações sanguíneas de AGCC totais e acetato nas fezes, porém não no sangue nas dietas a base de FO. A glicemia e insulinêmica de asininos é menor do que a de equinos. Dietas com açúcar e amido apresentam maiores níveis glicêmicos e insulinêmicos para ambas as espécies. Dietas a base de fibra e óleo se mostraram promissoras para ambas as espécies, em especial para os asininos. / The aim of the present study was to evaluate the inclusion of two energy sources (high sugar and high starch (AA), high fiber and high oil (FO)) in the diet of asinines and equines, through the trials of apparent digestibility of nutrients, physico-chemical characteristics of feaces and blood parameters, related to equine energy metabolism. Twenty equids, 10 Quarter Horse horses and 10 Pêga breed donkeys were used, distributed in an alternate Cross-Over type design, with two energetic sources and two species. Experimental diets were composed of 50% of the energy from the Tifton 85 (Cynodon spp.) and 50% from the concentrate, based on starch and sugar, and fiber and oil. It was observed effect of diet (p&lt;0.05) on the digestibility coefficients of ether extract and acid detergent fiber, buffer capacity at pH 5 and pH 6 (higher values for fiber and oil diet) and on blood concentrations of glucose (higher values for sugar and starch diet). It was observed effect of specie (p&lt;0.05) on the digestibility coefficient of crude protein, on buffering capacities at pH 5 and on blood concentrations of glucose (higher glycemic levels observed for equines). There was effect of the collection time (p&lt;0.05) on the plasmatic concentrations of acetate. For blood and fecal concentrations of short chain fatty acids, there was no effect for propionate (p&lt;0.05) of diet or specie. Diets composed by predominance of forage plus concentrate based on starch and sugar, and fiber and oil are equally used by the equids with emphasis of the fiber and oil diet in the use of lipids, without causing deleterious effect on the physico-chemical characteristics, pH and buffer capacity of feaces. Donkeys are more effective in buffering intestinal acids and also have higher blood concentrations of total SCFA and acetate in faeces, but not in blood in FO-based diets.

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No. of bitstreams: 1 ArianaLCP_TESE.pdf: 1354175 bytes, checksum: 3b6daaceb10d7ed6b27d02cbd71cb2c9 (MD5) Previous issue date: 2016-07-29 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / We aimed to assess whether cortisol and the physiological variables of asinine are influenced by meteorological variables, and if tramadol at doses of 2.5mg.kg-1 and 4mg.kg-1 IV promotes post-surgical analgesia in donkeys that have undergone orchiectomy. A priori, we used 10 north-eastern donkeys, in which we evaluated heart (HR) and respiratory (fR) rates, systolic (SBP), diastolic (DBP) and mean (MBP) blood pressure, rectal temperature (RT) and cortisol. The weather station of UFERSA provided temperature of the environment (TE), relative humidity (RH) and wind speed (WS) values, from which we calculated the temperature and humidity index (THI) every 2 hours for 24 hours. Afterwards, 24 donkeys, divided into 3 groups of 8 animals each: CG (control group, saline IV), GT2.5 (2.5 mg.kg-1 of tramadol IV) and GT4.0 (4.0 mg.kg-1 of tramadol IV) either received or did not analgesic after castration. The physiological variables mentioned above were re-evaluated and inserted in a multifactorial numerical scale to assess the analgesia within 1h, 2h, 4h, 6h, 8h, 12h and 24h after surgery. Statistical differences between groups in each hour and between hours for each particular group were obtained through the Kruskal-Wallis and Friedman tests, respectively. The relationship between the physiological and meteorological variables were analysed through the Spearman correlation. In the cortisol and physiological variables applied Principal Component Analysis (PCA). HR did not change significantly. The RT rose from 6h until 18h, the fR until 20h, and the blood pressures peaked at 2h and 14h. TE rose significantly from 6h to 14h, the WS until 12h, while the RH decreased until 12h. From 8h until 24h THI was in the range of 71-78, considered critical, its highest average occurred at 14h. There was significant negative correlation between fR and WS, as well as between RT and WS; and positive between fR and RH. Cortisol increased at 6h, peaking every 6h. The mean pain scores in the tramadol groups were significantly lower than those in the CG at 4h after surgery. In total, 87.5% of the GT4.0 animals received rescue analgesia, against 100% of the other groups. We concluded that the meteorological variables influence the physiological variables of donkeys in the north-eastern semi-arid. Tramadol at doses of 2.5 and 4.0 mg.kg-1 IV increased postsurgical analgesia for a period of 4-6h in donkeys that had undergone castration. The doses used did not cause deleterious changes to the cardiovascular parameters of the asinine / Objetivou-se avaliar se o cortisol e os parâmetros fisiológicos de asininos são influenciados pelas variáveis meteorológicas, e se o tramadol nas doses de 2,5 mg.kg-1 e 4 mg.kg-1IV promove analgesia pós-cirúrgica em asininos submetidos à orquiectomia. À priori, utilizaramse 10 animais, nos quais avaliaram-se frequências cardíaca (FC) e respiratória (fR), pressão arterial sistólica (PAS), diastólica (PAD) e média (PAM), temperatura retal (TR) e cortisol. A estação meteorológica da UFERSA forneceu os dados de temperatura ambiente (TA), umidade relativa do ar (UR) e velocidade do vento (VV), a partir daí calculou-se o índice de temperatura e umidade (ITU) a cada 2h, durante 24 horas. À posteriori, 24 asininos, divididos em 3 grupos de 8 animais: GC (grupo controle, solução salina IV), GT2,5 (tramadol 2,5 mg.kg-1 IV) e GT4,0 (tramadol 4,0 mg.kg-1 IV) receberam ou não analgésico após castração. Os parâmetros fisiológicos acima citados foram reavaliados e inseridos em escala numérica multifatorial para avaliação da analgesia em 1h, 2h, 4h, 6h, 8h, 12h e 24h após a cirurgia. Diferenças estatísticas entre grupos dentro de cada momento e entre momentos para cada grupo, foram testadas com Kruskall-Wallis e Friedman respectivamente, as relações existentes entre as variáveis fisiológicas e meteorológicas, correlação de Spearman. Para cortisol e parâmetros fisiológicos aplicou-se Análise de Componentes Principais (ACP). A FC não variou significativamente. A TR elevou-se das 6h até às 18h, a fR até às 20h, e as pressões apresentaram picos às 2h e às 14h. A TA elevou-se significativamente das 6h até às 14h, a VV até às 12h, enquanto que a UR diminuiu até às 12h. De 8h às 24h o ITU estava na faixa de 71- 78, a maior média ocorreu às 14h. Houve correlação significativa negativa entre fR e a VV, e da TR com a VV; e positiva entre fR a com UR. O cortisol apresentou elevação às 6h da manhã, com picos a cada 6h. As médias dos escores de dor dos grupos tramadol foram significativamente menores do que o GC em 4h após a cirurgia. No total, 87,5% dos animais do GT4,0 receberam analgesia resgate, contra 100% dos demais grupos. Concluiu-se que as variáveis meteorológicas influenciam os parâmetros fisiológicos de asininos do semiárido nordestino. O tramadol nas doses de 2,5 e 4,0 mg.kg-1IV incrementaram a analgesia póscirúrgica pelo período de 4-6h em asininos submetidos à castração. As doses utilizadas não causaram alterações deletérias aos parâmetros cardiovasculares de asininos / 2017-04-18

Made available in DSpace on 2015-03-26T12:54:57Z (GMT). No. of bitstreams: 1 texto completo.pdf: 1973320 bytes, checksum: aa4f936802e91a3fcfd777d09de0f494 (MD5) Previous issue date: 2013-10-11 / We aimed with this study to evaluate the effect of photoperiod changes on reproductive characteristics of Pêgabreed jennies along the year and to verify the embryo recovery rate and embryo characteristics on days 7, 8 and 9 postovulation. In Experiment 1, 279 estrous cycles of 44 jennies were evaluated during 3 consecutive years. The experimental groups were split in two periods according to photoperiod along the year in the southern hemisphere: P1: October to March (Long photoperiod); P2: April to September (Short photoperiod). The sexual behavior, ovarian and uterine characteristics, and fertility during 3 years were evaluated. Jennies were checked for estrous behavior by a jack and were evaluated by transrectal palpation every two days to identify females in estrous. Jennies with sexual signs of estrous or with anovarian follicle bigger than 25 mm in diameter were controlled daily until ovulation was detected.Artificial insemination or natural breed was performed every two days when a follicle reached &#8805; 35 mm until ovulation. Pregnancy diagnose was done on days 13, 25 and 35 postovulation by ultrasonography using a transrectal linear transducer (5 MHz). Differences were not detected (P>0.05) between the two periods in the following characteristics: follicle diameter at the beginning of estrous (29.80 ± 0.18mm), follicle diameter at ovulation (44.61± 0.24mm), estrous length (7.44± 0.05days),interval from the onset of estrous to ovulation (5.99± 0.06days), interval from ovulation to the estrous end (1.45± 0.03days), follicle growth rate (2.50± 0.04mm/day)frequency of double (16.49%, 46/279) and triple (1.43%, 4/279)ovulations, frequency of ovulation in the left (52.65%, 146/279)and right (47,35%, 133/279)ovaries, and frequency of ovulations with visual signs in the pre-ovulatory follicle (77.85%, 217/279).Total pregnancy rate was 60%; a difference in pregnancy rate between the two periods was not detected (P>0.1).Photoperiod variation did not affect the characteristics of follicle growth, sexual behavior and fertility in jennies of Pêga breed under the present environment conditions.In Experiment 2, the recovery rate, development and quality of embryos on days 7, 8 and 9 were evaluated. For embryo recovery, 90 uterine flushes were performed in seven Pêga breed jennies, aging 6 to 14 years old. Sixty-three embryos from 90 flushes (70%) and 107 ovulations (58.9%) were recovered. Averaged embryo diameter was 747.38 ± 76.45 &#956;m. The proportions of embryos development stages were as follow: morulas,1.59% (1/63);early blastocysts, 6.35% (4/63);blastocysts, 23.81% (15/63); and expanded blastocysts,68.25% (43/63). For quality grades, 90.5% (57/63) of embryos were classified as excellent or good (Grade I and II) and 9.5% (6/63) as regular or poor (Grade III and IV). No difference (P>0.1) in embryo recovery rate was detected between the two year periods. The embryo quality observed in the present study is considered satisfactory in a embryo transfer program. / Objetivou-se nesse estudo avaliar o efeito da variação do fotoperíodo sobre as características reprodutivas de jumentas da raça Pêga ao longo do ano; verificar a taxa de recuperação e viabilidade embrionária de lavados feitos nos dias 7, 8 e 9 pós-ovulação. No Experimento 1 foram avaliados 279 ciclos de 44 jumentas durante 3 anos consecutivos, sendo os grupos experimentais divididos de acordo com os períodos do ano: P1: outubro a março (período de fotoperíodo mais longo); P2: abril a setembro (período de fotoperíodo mais curto). Foram avaliadas as características de comportamento sexual, características ovarianas, uterinas e fertilidade. As jumentas foram rufiadas e avaliadas por palpação retal e ultrassonografia a cada dois dias. Aquelas que apresentavam comportamento de estro ou a presença de folículo maior que 25 mm de diâmetro em um dos ovários, foram controladas diariamente até a ovulação. Após a detecção de um folículo maior ou igual a 35 mm de diâmetro as jumentas foram artificialmente inseminadas ou cobertas a cada 48 horas até a ovulação. Os diagnósticos de gestação foram realizados aos 13, 25 e aos 35 dias após a ovulação por ultra-sonografia utilizando transdutor linear trans-retal de 5 MHz. Os valores médios das características diâmetro folicular no início do estro, diâmetro folicular pré- ovulação, duração do estro, intervalo início do estro à ovulação, intervalo da ovulação ao fim do estro, taxa de crescimento folicular, foram de 29,80±0,18 mm, 44,61±0,24 mm, 7,44±0,05 dias, 5,99±0,06 dias, 1,45±0,03 dias, 2,50±0,04 mm/dia, respectivamente. A porcentagem de duplas e triplas ovulações, frequência de ovulações entre os ovários direito e esquerdo e porcentagem de ovulações com alterações do folículo pré-ovulatório foram de 16,49% (46/279), 1,43% (4/279), 52,65% (146/279), 47,35% (133/279) e 77,85% (217/279), respectivamente. A taxa de prenhez geral foi de 60%. Não foram observadas entre os períodos, diferenças (P>0,05) nas características avaliadas. A variação do fotoperíodo não afetou as características foliculares, comportamento sexual e fertilidade de jumentas da raça Pêga nas condições experimentais avaliadas. No Experimento 2 avaliou-se a taxa de recuperação, as características morfométricas e estádio de desenvolvimento embrionário de embriões coletados nos dias D7, D8 e D9. Foram realizados 90 lavados uterinos de 7 jumentas da raça Pêga com idade variando de 6 a 14 anos. Foram recuperados 63 embriões de 90 lavados (70%) referentes a 107 ovulações (58,9%). O diâmetro médio dos embriões foi de 747.38 ± 76.45 &#956;m. Dentre os embriões colhidos, as mórulas corresponderam a 1,59% (1/63), os blastocistos iniciais a 6,35% (4/63), os blastocistos a 23,81% (15/63) e os blastocistos expandidos 68,25% (43/63), dos embriões totais. Quanto à qualidade, 90,5% (57/63) dos embriões foram classificados como excelentes ou bons (Graus I e II) e 9,5% (6/63) como regulares ou ruins (Graus III e IV). Não foi observada diferença da taxa de recuperação embrionária entre os períodos do ano. Os dados de qualidade dos embriões obtidos no presente estudo são considerados satisfatórios para um programa de TE.

O objetivo do presente estudo foi avaliar a inclusão de duas fontes energéticas (alto açúcar e alto amido (AA), alta fibra e alto óleo (FO)) na dieta de asininos e equinos, através de ensaios de digestibilidade aparente dos nutrientes, características físico-químicas das fezes e parâmetros sanguíneos, vinculados ao metabolismo energético dos equídeos. Foram utilizados 20 equídeos, dez equinos da raça Quarto de Milha e dez asininos da raça Pêga, distribuídos no delineamento alternado do tipo Cross-Over, com duas fontes energéticas e duas espécies. As dietas experimentais foram compostas de 50% da energia proveniente do Feno Tifton 85 (Cynodon spp.) e 50% proveniente do concentrado, formulados a base amido e açúcar, e fibra e óleo. Foi observado efeito de dieta (p&lt;0,05) sobre os coeficientes de digestibilidade do extrato etéreo e fibra em detergente ácido, capacidade tamponante a pH 5 e pH 6 (maiores valores para dieta Fibra óleo) e sobre as concentrações sanguíneas de glicose (maiores valores para dietas açúcar amido. Observou-se efeito de espécie (p&lt;0,05) sobre o coeficiente de digestibilidade aparente de proteína bruta, sobre a capacidades tamponante à pH 5 e sobre as concentrações sanguíneas de glicose (maiores níveis glicêmicos observados para equinos). Houve efeito de tempo de coleta (p&lt;0,05) sobre as concentrações plasmáticas de acetato. Para as concentrações sanguíneas e fecais de ácidos graxos de cadeia curta totais, propionato não houve efeito (p&gt;0,05) de dieta ou de espécie. Dietas composta por predominância de volumoso acrescido de concentrado a base de amido e açúcar e fibra e óleo são igualmente aproveitadas pelos equídeos com destaque para a dieta fibra e óleo no aproveitamento de lipídios, sem causar efeito deletério sobre as características físico-químicas, o pH e a capacidade tamponante das fezes. Asininos são mais eficientes no tamponamento de ácidos intestinais. Os asininos apresentam maiores concentrações sanguíneas de AGCC totais e acetato nas fezes, porém não no sangue nas dietas a base de FO. A glicemia e insulinêmica de asininos é menor do que a de equinos. Dietas com açúcar e amido apresentam maiores níveis glicêmicos e insulinêmicos para ambas as espécies. Dietas a base de fibra e óleo se mostraram promissoras para ambas as espécies, em especial para os asininos. / The aim of the present study was to evaluate the inclusion of two energy sources (high sugar and high starch (AA), high fiber and high oil (FO)) in the diet of asinines and equines, through the trials of apparent digestibility of nutrients, physico-chemical characteristics of feaces and blood parameters, related to equine energy metabolism. Twenty equids, 10 Quarter Horse horses and 10 Pêga breed donkeys were used, distributed in an alternate Cross-Over type design, with two energetic sources and two species. Experimental diets were composed of 50% of the energy from the Tifton 85 (Cynodon spp.) and 50% from the concentrate, based on starch and sugar, and fiber and oil. It was observed effect of diet (p&lt;0.05) on the digestibility coefficients of ether extract and acid detergent fiber, buffer capacity at pH 5 and pH 6 (higher values for fiber and oil diet) and on blood concentrations of glucose (higher values for sugar and starch diet). It was observed effect of specie (p&lt;0.05) on the digestibility coefficient of crude protein, on buffering capacities at pH 5 and on blood concentrations of glucose (higher glycemic levels observed for equines). There was effect of the collection time (p&lt;0.05) on the plasmatic concentrations of acetate. For blood and fecal concentrations of short chain fatty acids, there was no effect for propionate (p&lt;0.05) of diet or specie. Diets composed by predominance of forage plus concentrate based on starch and sugar, and fiber and oil are equally used by the equids with emphasis of the fiber and oil diet in the use of lipids, without causing deleterious effect on the physico-chemical characteristics, pH and buffer capacity of feaces. Donkeys are more effective in buffering intestinal acids and also have higher blood concentrations of total SCFA and acetate in faeces, but not in blood in FO-based diets.

Les pathologies intestinales sont des maladies multifactorielles dont l’incidence ne cesse d’augmenter. Les maladies organiques regroupent les maladies inflammatoires chroniques de l'intestin tel que la maladie de Crohn (MC) et les pathologies fonctionnelles digestives comprennent les différentes formes du syndrome de l'intestin irritable (SII). Ces pathologies présentes des caractéristiques communes comme des troubles du transit, des douleurs abdominales, un dysfonctionnement de la barrière intestinale et des modifications de la communication bidirectionnelle de l’axe intestin-cerveau et une altération de la composition du microbiote intestinal (dysbiose). Cette dernière peut être en partie associée à un déficit de production en peptides et protéines antimicrobiennes (PAMs) par les cellules de Paneth. Parmi les cibles thérapeutiques stratégiques, une réduction de la dysbiose dans le but de réduire l’état inflammatoire ou micro-inflammatoire de la muqueuse font aujourd’hui l’objet de différentes études. Ces traitements alternatifs montrent l’efficacité d’un régime nutritionnel adapté, d’un apport en probiotiques ou en prébiotiques sur le microbiote intestinal des patients MC et SII, et sur l’intégrité de leur barrière intestinale. Parmi les matrices alimentaires présentant une composition nutritionnelle intéressante proche du lait maternel humain, le lait d’ânesse (LA) contient également une forte teneur en PAMs (lysozyme). En parallèle, un observatoire économique et social de la filière asine commandité par l’Institut National Âne et Mulet fait le diagnostic d’une activité grandissante autour du LA chez les éleveurs français et montre, face à une concurrence italienne et chinoise importante, la nécessité d’une meilleure organisation de la filière via une meilleure valorisation du LA. L’objectif de cette thèse était d’évaluer un potentiel effet santé du LA et d’apporter des preuves scientifiques robustes pour permettre une meilleure valorisation de ce dernier. Ainsi, l’effet d’une consommation orale chronique de LA a été évalué sur deux modèles précliniques murins distincts : un modèle d’iléite expérimentale induite par une administration per os d’indométacine et un modèle de stress psychologique chronique (Water Avoidance Stress, WAS). Dans ce travail, le rôle clé de l’activité du lysozyme dans les effets observés a également été évalué. Des essais de traitements thermiques ont aussi été réalisés pour optimiser un barème de pasteurisation permettant de proposer un LA répondant aux normes réglementaires microbiologiques et sanitaires tout en conservant l’activité du lysozyme contenue dans le lait. Les travaux de cette thèse montrent que le LA possède des propriétés anti-inflammatoires se traduisant par une réduction significative des lésions inflammatoires macroscopiques et microscopiques de l’iléon. Cet effet est associé à une réduction de la dysbiose intestinale et normalise les niveaux de PAMs dans les cellules de Paneth drastiquement réduits lors de l’iléite. Dans le modèle WAS, le LA et la fraction contenant l’activité en lysozyme réduisent l’hypersensibilité viscérale, l’état micro-inflammatoire induites par le stress et restaurent les niveaux de PAMs dans les cellules de Paneth réduits par le stress. Un traitement thermique de 2 min/72°C permet d’augmenter la durée de vie du LA tout en conservant l’activité du lysozyme et ses propriétés santé sur la muqueuse intestinale chez la souris. / Gut pathologies are multifactorial with a constant increasing incidence. Organic diseases include inflammatory bowel diseases such as Crohn's disease (CD) and functional pathologies include various subtypes of irritable bowel syndrome (IBS). These pathologies present common features such as transit disorders, abdominal pain, dysfunction of the intestinal barrier associated with bidirectional gut-brain axis modifications and alterations of the gut microbiota composition (dysbiosis). Dysbiosis is generally associated with a lack of production of antimicrobial peptides and proteins (AMPs) by Paneth cells. Among the alternative treatment strategies, it has been proposed to reduce gut inflammation by correcting intestinal dysbiosis. Recent studies show the efficacy of a suitable diet, probiotics or prebiotics interventions on gut microbiota and on the integrity of their intestinal barrier in CD and IBS patients. Among interesting food matrices, donkey milk (DM) is a good candidate with a nutritional composition close to human breast milk and contains high levels of AMPs like lysozyme. An economic and social observatory of the donkey industry was ordered by the Institut National Âne et Mulet for a better support of growing French activity around DM against Italian and Chinese competitions. The objective of this thesis was to evaluate the potential beneficial healthy effect of DM and to provide robust scientific evidences for a better valorization of this product. Thus, the effect of chronic oral intake of DM was evaluated on two distinct murine preclinical models: a model of experimental ileitis induced by an oral administration of indomethacin and a model of chronic psychological stress (Water Avoidance Stress, WAS). In these studies, the key role of lysozyme activity in the observed effects was highlighted. In order to develop the DM market in France, thermal treatments have been carried out to optimize a pasteurization process to obtain a DM in accordance with the regulatory microbiological and sanitary standards while preserving the activity of the lysozyme contained in milk. The work of this thesis shows that DM exerts anti-inflammatory properties resulting in a significant reduction of the macroscopic and microscopic inflammatory lesions of the ileum. This effect is associated with a reduction of gut dysbiosis while normalizing the level of drastically reduced AMPs contained Paneth cells in the ileitis model. In the WAS model, DM and the fraction containing activity of lysozyme reduce visceral hypersensitivity, stress-induced gut microinflammatory status and also restore the level of AMPs in Paneth cells . A heat treatment at 2 min/72 °C allows to increase the shelf-life of DM while preserving the activity of lysozyme and its gut beneficial healthy properties in mouse.

<p>Млеко магарице је тек последњих година доспело у фокус научних истраживања, иако су његова благотоворна терапеутска и козметичка својства позната још од античких времена. Упркос томе што знање о функционалности млека магарице константно расте, његов антимикробни потенцијал остаје најмање истражен. Литературни подаци о микробиологији и евентуалној антимикробној активности млека магарице балканске расе која егзистира на подручју Србије и околних балканских земаља су јако лимитирани. Ово млеко се традиционално конзумира без претходне топлотне обраде, што покреће низ питања која се односе на здравствену безбедност конзумирања млека у сировом стању. Предмети истраживања ове докторске дисертације били су утврђивање микробиолошког квалитета сировог млека магарице балканске расе, као и испитивање његове антибактеријске активности према одабраним сојевима Escherichia coli, Salmonella Enteritidis, Salmonella Typhimurium, Staphylococcus aureus, Listeria monocytogenes и Klebsiella pneumoniae. У циљу утврђивања главних носилаца антибактеријског дејства, као и самог модела антибактеријског деловања, у одабраним узорцима овог млека намењеним извођењу антибактеријских испитивања, извршено је одређивање садржаја антимикробних протеина - лизозима и лактоферина, као и садржаја калцијума. У узорцима млека магарице предвиђеним за испитивање антибактеријске активности према S. aureus и L. monocytogenes, одређен је садржај масних киселина са документованом антибактеријском активношћу према Грам<br />позитивним бактеријама. Присуство L. monocytogenes и Salmonella spp. није утврђено ни у једном од 137 испитаних узорака сировог млека магарице, док је број E. coli, колиформних бактерија, Enterobacteriaceae, квасаца и плесни и суспектног B. cereus био испод границе детекције методе. Према резултатима добијеним у овој докторској дисертацији сви испитани узорци сировог млека магарице балканске расе су задовољавали захтеве европских регулатива 92/46/ЕЕC и EC 853/2004 и са микробиолошког аспекта су у потпуности били безбедни за директно конзумирање без претходне топлотне обраде. Сирово млеко магарице балканске расе је показало различиту антибактеријску активност према Грам позитивним и Грам негативним бактеријским сојевима тестираним у оквиру ове докторске дисертације, чији интензитет је зависио од нивоа контаминације узорака млека тестираним патогеном и температуре инкубирања. Температура инкубирања од +15 &deg;C је у односу на +9 &deg;C била генерално повољнија за активност антибактеријских супстанци у млеку магарице према свим тестираним патогенима, али је антибактеријски потенцијал узорака млека магарице на +15 &deg;C био лимитиран бржим умножавањем испитаних бактерија на овој температури. То је посебно важило за узорке са већим нивоима контаминације (103 и 104 cfu/ml). Антибактеријска активност сировог млека магарице према L. monocytogenes и S. aureus се нa свим радним температурама састојала од продужeња лаг фазе и инхибирања бактеријског раста, док се у случају S. Enteritidis, S. Typhimurium, E. coli АTCC 8739, E. coli АTCC 10536 и K. pneumoniae она огледала у редукцији њиховог почетног броја и/или продужењу лаг фазе. Резултати ове докторске дисертације су указали на калцијум зависну природу антибактеријске активности сировог млека магарице према S. Enteritidis, S. Typhimurium и оба тестирана соја E. coli. Најснажнију антибактеријску активност према K. pneumoniae показали су узорци млека магарице, који су поред великог садржаја лизозима имали и велики садржај лактоферина, па би у овом случају основни механизам антибактеријског деловања млека магарице могао бити синергизам лизозима и лактоферина. Лизозим је највероватније главни носилац антибактеријске активности млека магарице балканске расе, обзиром да је у односу на други антимикробни агенс &ndash; лактоферин у тестираним узорцима млека магарице детектован у вишеструко већој концентрацији. Известан допринос антибактеријској активности млека<br />магарице балканске расе према L. monocytogenes и S. aureus могле би да дају и масне киселине са документованим антибактеријским дејством према Грам позитивним бактеријама (линолна, лауринска и олеинска киселина), које су у анализираним узорцима млека магарице у збиру чиниле од 40,3 до 54,7% од укупних масних киселина.</p> / <p>Mleko magarice je tek poslednjih godina dospelo u fokus naučnih istraživanja, iako su njegova blagotovorna terapeutska i kozmetička svojstva poznata još od antičkih vremena. Uprkos tome što znanje o funkcionalnosti mleka magarice konstantno raste, njegov antimikrobni potencijal ostaje najmanje istražen. Literaturni podaci o mikrobiologiji i eventualnoj antimikrobnoj aktivnosti mleka magarice balkanske rase koja egzistira na području Srbije i okolnih balkanskih zemalja su jako limitirani. Ovo mleko se tradicionalno konzumira bez prethodne toplotne obrade, što pokreće niz pitanja koja se odnose na zdravstvenu bezbednost konzumiranja mleka u sirovom stanju. Predmeti istraživanja ove doktorske disertacije bili su utvrđivanje mikrobiološkog kvaliteta sirovog mleka magarice balkanske rase, kao i ispitivanje njegove antibakterijske aktivnosti prema odabranim sojevima Escherichia coli, Salmonella Enteritidis, Salmonella Typhimurium, Staphylococcus aureus, Listeria monocytogenes i Klebsiella pneumoniae. U cilju utvrđivanja glavnih nosilaca antibakterijskog dejstva, kao i samog modela antibakterijskog delovanja, u odabranim uzorcima ovog mleka namenjenim izvođenju antibakterijskih ispitivanja, izvršeno je određivanje sadržaja antimikrobnih proteina - lizozima i laktoferina, kao i sadržaja kalcijuma. U uzorcima mleka magarice predviđenim za ispitivanje antibakterijske aktivnosti prema S. aureus i L. monocytogenes, određen je sadržaj masnih kiselina sa dokumentovanom antibakterijskom aktivnošću prema Gram<br />pozitivnim bakterijama. Prisustvo L. monocytogenes i Salmonella spp. nije utvrđeno ni u jednom od 137 ispitanih uzoraka sirovog mleka magarice, dok je broj E. coli, koliformnih bakterija, Enterobacteriaceae, kvasaca i plesni i suspektnog B. cereus bio ispod granice detekcije metode. Prema rezultatima dobijenim u ovoj doktorskoj disertaciji svi ispitani uzorci sirovog mleka magarice balkanske rase su zadovoljavali zahteve evropskih regulativa 92/46/EEC i EC 853/2004 i sa mikrobiološkog aspekta su u potpunosti bili bezbedni za direktno konzumiranje bez prethodne toplotne obrade. Sirovo mleko magarice balkanske rase je pokazalo različitu antibakterijsku aktivnost prema Gram pozitivnim i Gram negativnim bakterijskim sojevima testiranim u okviru ove doktorske disertacije, čiji intenzitet je zavisio od nivoa kontaminacije uzoraka mleka testiranim patogenom i temperature inkubiranja. Temperatura inkubiranja od +15 &deg;C je u odnosu na +9 &deg;C bila generalno povoljnija za aktivnost antibakterijskih supstanci u mleku magarice prema svim testiranim patogenima, ali je antibakterijski potencijal uzoraka mleka magarice na +15 &deg;C bio limitiran bržim umnožavanjem ispitanih bakterija na ovoj temperaturi. To je posebno važilo za uzorke sa većim nivoima kontaminacije (103 i 104 cfu/ml). Antibakterijska aktivnost sirovog mleka magarice prema L. monocytogenes i S. aureus se na svim radnim temperaturama sastojala od produženja lag faze i inhibiranja bakterijskog rasta, dok se u slučaju S. Enteritidis, S. Typhimurium, E. coli ATCC 8739, E. coli ATCC 10536 i K. pneumoniae ona ogledala u redukciji njihovog početnog broja i/ili produženju lag faze. Rezultati ove doktorske disertacije su ukazali na kalcijum zavisnu prirodu antibakterijske aktivnosti sirovog mleka magarice prema S. Enteritidis, S. Typhimurium i oba testirana soja E. coli. Najsnažniju antibakterijsku aktivnost prema K. pneumoniae pokazali su uzorci mleka magarice, koji su pored velikog sadržaja lizozima imali i veliki sadržaj laktoferina, pa bi u ovom slučaju osnovni mehanizam antibakterijskog delovanja mleka magarice mogao biti sinergizam lizozima i laktoferina. Lizozim je najverovatnije glavni nosilac antibakterijske aktivnosti mleka magarice balkanske rase, obzirom da je u odnosu na drugi antimikrobni agens &ndash; laktoferin u testiranim uzorcima mleka magarice detektovan u višestruko većoj koncentraciji. Izvestan doprinos antibakterijskoj aktivnosti mleka<br />magarice balkanske rase prema L. monocytogenes i S. aureus mogle bi da daju i masne kiseline sa dokumentovanim antibakterijskim dejstvom prema Gram pozitivnim bakterijama (linolna, laurinska i oleinska kiselina), koje su u analiziranim uzorcima mleka magarice u zbiru činile od 40,3 do 54,7% od ukupnih masnih kiselina.</p> / <p>Donkey&#39;s milk has recently become a very popular topic among scientific community although it has been used for therapeutic and cosmetic purposes since ancient times. Even though the number of investigations concerning donkey&#39;s milk functionality has increased, the studies on its antimicrobial potential are limited. Especially, there is a lack in literature data on microbiological quality and antibacterial activity of donkey&#39;s milk obtained from Domestic Balkan breed originated in Serbia and other countries in Balkan region. The fact that this milk is traditionally consumed without heat treatment launches a series of issues concerning its safety.<br />The aim of this thesis was to determine microbiological quality of Domestic Balkan donkey&#39;s milk, as well as to investigate its antibacterial activity against the selected strains of Escherichia coli, Salmonella Enteritidis, Salmonella Typhimurium, Staphylococcus aureus, Listeria monocytogenes and Klebsiella pneumonia. In order to determine the major antibacterial substances, as well as the model of antibacterial activity, the content of antimicrobial proteins (lysozyme and lactoferrin) and calcium was determined. The amount of fatty acids which are known to be effective against Gram-positive bacteria was determined in milk samples tested for antibacterial activity against S. aureus и L. monocytogenes. The presence of L. monocytogenes and Salmonella spp. was not detected in any of the 137 raw donkey&#39;s milk samples, while the levels of E. coli, coliform bacteria, Enterobacteriaceae, yeast and molds and suspect Bacillus cereus were below the limit of detection of the assays. According to obtained results, all examined Domestic Balkan donkeys&#39; raw milk samples were<br />in accordance with the EU regulations 92/46/ЕЕC and EC 853/2004 in terms of their<br />microbiological quality, thus being completely safe for direct consumptions, without thermal treatment. Raw Domestic Balkan donkey&#39;s milk has expressed different antibacterial activities against Gram-positive and Gram-negative bacteria strains tested in this thesis, with the intensities which depended on contamination level and incubation temperature. In comparison to temperature of +9 &deg;C, the incubation temperature of +15 &deg;C was more favourable for activity of antibacterial substances in donkey&#39;s milk against all tested pathogens. However, the antibacterial potential of donkey&#39;s milk samples at +15 &deg;C was limited with rapid multiplication of bacteria at this temperature. This was especially pronounced in samples characterized with higher contamination levels (103 and 104 cfu/ml). Antibacterial activity of raw donkey&#39;s milk against L. monocytogenes and S. aureus at all tested temperatures reflected in prolonged lag phase and inhibition of bacterial growth, while the activity against S. Enteritidis, S. Typhimurium, E. coli АTCC 8739, E. coli АTCC 10536 and K. pneumonia comprised reduction of initial bacterial numbers and/or prolongation of lag phase.<br />Results obtained in this thesis have indicated on calcium dependent antibacterial activity of raw donkey&#39;s milk against S. Enteritidis, S. Typhimurium and both E. coli strains. The most pronounced antibacterial activity against K. pneumonia was found in donkey&#39;s milk samples which besides high content of lysozyme had high content of lactoferrin, thus indicating that the basic antibacterial mechanism of donkey&#39;s milk might be the synergism between lysozyme and lactoferrin. Lysozyme was probably the substance with the highest antimicrobial activity in Domestic Balkan donkey&#39;s milk, since it was detected in significantly higher concentrations than lactoferrin. The fatty acids which are known to be effective against Gram-positive bacteria (linoleic, lauric and oleic) and which comprised from 40.3 to 54.7% of total fatty acids in tested samples, may also contribute to antimicrobial activity of Domestic Balkan donkey&#39;s milk against L. monocytogenes and S. aureus.</p>