Silagens de grãos de milho puro e com adição de grãos de soja, de girassol ou uréia

O objetivo deste trabalho foi avaliar a composição química e a estabilidade em aerobiose de silagens de grãos de milho, ensilados com adição de soja crua, girassol ou uréia. Em delineamento inteiramente casualizado, com quatro repetições, foram avaliados os tratamentos: grãos de milho; grãos de milho mais grãos de soja; grãos de milho mais grãos de girassol; e grãos de milho mais uréia. Foram utilizados, como silos experimentais, tonéis de plástico com capacidade para 200 kg, que permaneceram vedados por nove meses. Avaliaram-se: a composição química, a degradabilidade ruminal da matéria seca e da proteína bruta, o desaparecimento do amido no rúmen e a estabilidade aeróbia das silagens. Todos os tratamentos tiveram efeito sobre a degradabilidade efetiva da matéria seca e da proteína bruta, e também sobre a composição química das silagens, especialmente nos teores de proteína bruta e extrato etéreo. A adição de 20% de grãos de soja, de girassol ou uréia (1%), na silagem de grãos de milho, melhora sua composição químico-bromatológica, porém reduz a degradabilidade efetiva da matéria seca e da proteína bruta. A adição de uréia à silagem de grãos úmidos de milho diminui as perdas de matéria seca e melhora a estabilidade em aerobiose.

Processing of metacaspase into a cytoplasmic catalytic domain mediating cell death in Leishmania major.

Metacaspases are cysteine peptidases that could play a role similar to caspases in the cell death programme of plants, fungi and protozoa. The human protozoan parasite Leishmania major expresses a single metacaspase (LmjMCA) harbouring a central domain with the catalytic dyad histidine and cysteine as found in caspases. In this study, we investigated the processing sites important for the maturation of LmjMCA catalytic domain, the cellular localization of LmjMCA polypeptides, and the functional role of the catalytic domain in the cell death pathway of Leishmania parasites. Although LmjMCA polypeptide precursor form harbours a functional mitochondrial localization signal (MLS), we determined that LmjMCA polypeptides are mainly localized in the cytoplasm. In stress conditions, LmjMCA precursor forms were extensively processed into soluble forms containing the catalytic domain. This domain was sufficient to enhance sensitivity of parasites to hydrogen peroxide by impairing the mitochondrion. These data provide experimental evidences of the importance of LmjMCA processing into an active catalytic domain and of its role in disrupting mitochondria, which could be relevant in the design of new drugs to fight leishmaniasis and likely other protozoan parasitic diseases.

Protozoários ciliados do rúmen de bovinos e bubalinos alimentados com dietas suplementadas com monensina ou própolis

O objetivo deste trabalho foi avaliar o efeito de dietas suplementadas com monensina ou produtos à base de própolis, nas populações de protozoários ciliados no rúmen de bovinos (Bos taurus) e bubalinos (Bubalus bubalis). Quatro bovinos da raça Holandesa e quatro búfalos da raça Murrah adultos, fistulados no rúmen, foram distribuídos em delineamento quadrado latino (4x4). A dieta constituiu-se de 50% de silagem de milho e 50% de concentrado à base de milho em grãos e farelo de soja, com adição de monensina sódica ou aditivo à base de própolis LLOSA2 ou LLOSC1. As amostras do conteúdo ruminal foram coletadas duas horas após a alimentação. O gênero Entodinium foi o mais representativo em todos os tratamentos, para ambas as raças de ruminantes. Em búfalos, foi observado o efeito redutor do tratamento LLOSC1 nas populações do gênero Entodinium, além do efeito redutor dos tratamentos monensina e LLOSA2 sobre os gêneros da subfamília Diplodiniinae. A média de ciliados foi maior em bubalinos (56x10(4) mL-1) do que em bovinos (26x10(4) mL-1). Houve aumento do pH ruminal dos bovinos no tratamento com monensina. O extrato de própolis LLOSC1 reduziu os ciliados do rúmen em bubalinos.

Suplementos para ovinos mantidos em pastos de capim-marandu

O objetivo deste trabalho foi avaliar o efeito da suplementação na resposta produtiva, no pH e no nitrogênio amoniacal ruminal, e no custo de produção de ovinos em pastagem de capim-marandu no período da seca. O experimento teve a duração de 84 dias. Utilizaram-se 20 cordeiros não castrados com idade e peso corporal inicial médios de quatro meses e 24,20 kg, respectivamente, para avaliação do desenvolvimento, distribuídos em cada um dos tratamentos que são os suplementos, mineral, energético, proteico e múltiplo, com cinco animais por tatamento, em área de 0,1 ha. Para avaliação dos parâmetros nutricionais, foram utilizados quatro ovinos fistulados no rúmen com 12 meses e 55 kg de peso corporal foram distribuídos em quatro piquetes de 0,1 ha. Avaliaram-se os suplementos mineral, energético, múltiplo e proteico. Os ganhos de peso foram de 0,017, -0,008, 0,024 e 0,077 kg por dia para os suplementos mineral, energético, múltiplo e proteico, respectivamente. Quatro horas após suplementação, os valores de pH ruminal foram de 6,30, 6,40, 6,18 e 6,24 para os suplementos mineral, energético, múltiplo e proteico, respectivamente. Os valores para nitrogênio amoniacal do líquido ruminal foram de 10,57, 7,36, 21,58 e 24,50 mg dL-1 para os suplementos mineral, energético, múltiplo e proteico, respectivamente. Os suplementos mineral e proteico produziram o ganho de peso com o menor custo. O uso de suplemento energético para cordeiros submetidos à forragem com baixo teor de proteína reduz o ganho de peso.

Tannin-tolerant bacteria from crossbred Holstein x Zebu cows

The objective of this work was to isolate and characterize tannin-tolerant ruminal bacteria from crossbred Holstein x Zebu cows fed a chopped mixture of elephant grass (Pennisetum purpureum), young stems of "angico-vermelho" (Parapiptadenia rigida), and banana tree (Musa sp.) leaves. A total of 117 bacteria strains were isolated from enrichment cultures of rumen microflora in medium containing tannin extracts. Of these, 11 isolates were able to tolerate up to 3 g L-1 of tannins. Classical characterization procedures indicated that different morphological and physiological groups were represented. Restriction fragments profiles using Alu1 and Taq1 of 1,450 bp PCR products from the 16S rRNA gene grouped the 11 isolates into types I to VI. Sequencing of 16S rRNA PCR products was used for identification. From the 11 strains studied, seven were not identifiable by the methods used in this work, two were strains of Butyrivibrio fibrisolvens, and two of Streptococcus bovis.

Parâmetros sanguíneos, hepáticos e ruminais de ovinos alimentados com dietas com farelo de mamona destoxificado

O objetivo deste trabalho foi avaliar os parâmetros sanguíneos, hepáticos e ruminais, bem como caracterizar e quantificar a população de protozoários ciliados, no rúmen de ovinos alimentados com dietas com farelo de mamona destoxificada (FM). Foram utilizados 32 ovinos, para a avaliação dos parâmetros sanguíneos e hepáticos, e quatro ovinos adultos fistulados no rúmen, para a avaliação dos parâmetros ruminais. A dieta controle foi composta por feno de capim-buffel, milho em grão moído, ureia e farelo de soja (FS). Nos tratamentos, o FS foi substituído parcialmente pelo FM a 15, 30 e 45% no concentrado. Não houve diferença, entre os tratamentos, quanto aos teores de ureia no soro, cuja média foi de 666,0 mg L-1. Não houve diferença entre as dietas quanto à glicose, ao aspartato aminotranferase e à alanina aminotransferase, que tiveram média de 690,3 mg L-1, 127,4 UI L-1 e 16,9 UI L-1, respectivamente. Os valores de nitrogênio amoniacal e pH apresentaram padrão linear crescente com a substituição do FS pelo FM. O gênero Entodinium foi o mais frequente em todos os tratamentos e obteve média geral de 76,4% do total de protozoários; o tratamento sem inclusão do FM obteve a maior percentagem deste gênero entre os tratamentos.Adieta com substituição de 45% do farelo de soja pelo de mamona destoxificado favorece o ambiente ruminal.

OmpA family proteins and Pmp-like autotransporter: new adhesins of Waddlia chondrophila.

Waddlia chondrophila is a obligate intracellular bacterium belonging to the Chlamydiales order, a clade that also includes the well-known classical Chlamydia responsible for a number of severe human and animal diseases. Waddlia is an emerging pathogen associated with adverse pregnancy outcomes in humans and abortion in ruminants. Adhesion to the host cell is an essential prerequisite for survival of every strict intracellular bacteria and, in classical Chlamydia, this step is partially mediated by polymorphic outer membrane proteins (Pmps), a family of highly diverse autotransporters that represent about 15% of the bacterial coding capacity. Waddlia chondrophila genome however only encodes one putative Pmp-like protein. Using a proteomic approach, we identified several bacterial proteins potentially implicated in the adhesion process and we characterized their expression during the replication cycle of the bacteria. In addition, we demonstrated that the Waddlia Pmp-like autotransporter as well as OmpA2 and OmpA3, two members of the extended Waddlia OmpA protein family, exhibit adhesive properties on epithelial cells. We hypothesize that the large diversity of the OmpA protein family is linked to the wide host range of these bacteria that are able to enter and multiply in various host cells ranging from protozoa to mammalian and fish cells.

Sanitary quality of edible bivalve mollusks in Southeastern Brazil using an UV based depuration system

The increase in seafood production, especially in mariculture worldwide, has brought out the need of continued monitoring of shellfish production areas in order to ensure safety to human consumption. The purpose of this research was to evaluate pathogenic protozoa, viruses and bacteria contamination in oysters before and after UV depuration procedure, in brackish waters at all stages of cultivation and treatment steps and to enumerate microbiological indicators of fecal contamination from production site up to depuration site in an oyster cooperative located at the Southeastern estuarine area of Brazil. Oysters and brackish water were collected monthly from September 2009 to November 2010. Four sampling sites were selected for enteropathogens analysis: site 1- oyster growth, site 2- catchment water (before UV depuration procedure), site 3 - filtration stage of water treatment (only for protozoa analysis) and site 4- oyster's depuration tank. Three microbiological indicators ! were examined at sites 1, 2 and 4. The following pathogenic microorganisms were searched: Giardia cysts, Cryptosporidium oocysts, Human Adenovirus (HAdV), Hepatitis A virus (HAV), Human Norovirus (HnoV) (genogroups I and II), JC strain Polyomavirus (JCPyV) and Salmonella sp. Analysis consisted of molecular detection (qPCR) for viruses (oysters and water samples); immunomagnetic separation followed by direct immunofluorescence assay for Cryptosporidium oocysts and Giardia cysts and also molecular detection (PCR) for the latter (oysters and water samples); commercial kit (Reveal-Neogee (R)) for Salmonella analysis (oysters). Giardia was the most prevalent pathogen in all sites where it was detected: 36.3%, 18.1%, 36.3% and 27.2% of water from sites 1, 2, 3 and 4 respectively; 36.3% of oysters from site 1 and 54.5% of depurated oysters were harboring Giardia cysts. The huge majority of contaminated samples were classified as Giardia duodenalis. HAdv was detected in water and o! ysters from growth site and HnoV GI in two batches of oysters ! (site 1) in huge concentrations (2.11 x 10(13), 3.10 x 10(12) gc/g). In depuration tank site, Salmonella sp., HAV (4.84 x 10(3)) and HnoV GII (7.97 x 10(14)) were detected once in different batches of oysters. Cryptosporidium spp. oocysts were present in 9.0% of water samples from site four. These results reflect the contamination of oysters even when UV depuration procedures are employed in this shellfish treatment plant. Moreover, the molecular comprehension of the sources of contamination is necessary to develop an efficient management strategy allied to shellfish treatment improvement to prevent foodborne illnesses. (C) 2011 Elsevier Ltd. All rights reserved.

Is Switzerland doing enough regarding the prevention of syphilis, gonorrhea and chlamydia infections ?

Sexually transmitted infections are a major problem for medicine and for public health services worldwide. More than 30 sexually transmittable pathogenic micro-organisms are known, including bacteria, viruses, fungi, protozoa and ectoparasites. According to estimates from the World Health Organisation more than 333 million of bacterial sexually transmitted infections occur worldwide per year. Sexually transmitted infections, by their nature, affect individuals, within partnerships and larger sexual networks, and in turn populations. This report focuses on three bacterial sexually transmitted infections in Switzerland that are Chlamydia trachomatis, Neisseria gonorrhea and Treponema pallidum (syphilis) in Switzerland. The prevalence of these infections has been increasing alarmingly for a decade. All three infections can be asymptomatic and their diagnosis and treatment can therefore occur too late or worse not at all, even though treatments are available. This is an important problem as untreated sexually transmitted infections may cause complications such as ascending infections, infertility, ectopic pregnancies and serious long-term neurological sequels. The consequences of these infections should not be underestimated. They constitute a significant public health burden as well as serious financial burden. The increases in chlamydia, syphilis and gonorrhea infections have also been observed in many European countries. Countries, where rising numbers of sexually transmitted infections have been observed, have reacted in different ways. Some have developed clinical guidelines or implemented screening programs, while others are still in their observational phase. The aim of this mémoire is to assess whether Switzerland is doing enough regarding the prevention of chlamydial, syphilis and gonorrheal infections. After first describing the infections, surveillance systems of sexually transmitted infections are assessed, then the epidemiological trends of these three infections are described, and finally the prevention measures implemented in Switzerland to respond to the increasing number of infections are described. The reaction of the United Kingdom to the same problem is reported for comparison. [Author, p. 7]

Importance of amoebae as a tool to isolate amoeba-resisting microorganisms and for their ecology and evolution: the Chlamydia paradigm.

Free-living amoebae are distributed worldwide and are frequently in contact with humans and animals. As cysts, they can survive in very harsh conditions and resist biocides and most disinfection procedures. Several microorganisms, called amoeba-resisting microorganisms (ARMs), have evolved to survive and multiply within these protozoa. Among them are many important pathogens, such as Legionella and Mycobacteria, and also several newly discovered Chlamydia-related bacteria, such as Parachlamydia acanthamoebae, Estrella lausannensis, Simkania negevensis or Waddlia chondrophila whose pathogenic role towards human or animal is strongly suspected. Amoebae represent an evolutionary crib for their resistant microorganisms since they can exchange genetic material with other ARMs and develop virulence traits that will be further used to infect other professional phagocytes. Moreover, amoebae constitute an ideal tool to isolate strict intracellular microorganisms from complex microbiota, since they will feed on other fast-growing bacteria, such as coliforms potentially present in the investigated samples. The paradigm that ARMs are likely resistant to macrophages, another phagocytic cell, and that they are likely virulent towards humans and animals is only partially true. Indeed, we provide examples of the Chlamydiales order that challenge this assumption and suggest that the ability to multiply in protozoa does not strictly correlate with pathogenicity and that we should rather use the ability to replicate in multiple and diverse eukaryotic cells as an indirect marker of virulence towards mammals. Thus, cell-culture-based microbial culturomics should be used in the future to try to discover new pathogenic bacterial species.

Implication of different domains of the Leishmania major metacaspase in cell death and autophagy.

Metacaspases (MCAs) are cysteine peptidases expressed in plants, fungi and protozoa, with a caspase-like histidine-cysteine catalytic dyad, but differing from caspases, for example, in their substrate specificity. The role of MCAs is subject to debate: roles in cell cycle control, in cell death or even in cell survival have been suggested. In this study, using a Leishmania major MCA-deficient strain, we showed that L. major MCA (LmjMCA) not only had a role similar to caspases in cell death but also in autophagy and this through different domains. Upon cell death induction by miltefosine or H2O2, LmjMCA is processed, releasing the catalytic domain, which activated substrates via its catalytic dyad His/Cys and a proline-rich C-terminal domain. The C-terminal domain interacted with proteins, notably proteins involved in stress regulation, such as the MAP kinase LmaMPK7 or programmed cell death like the calpain-like cysteine peptidase. We also showed a new role of LmjMCA in autophagy, acting on or upstream of ATG8, involving Lmjmca gene overexpression and interaction of the C-terminal domain of LmjMCA with itself and other proteins. These results allowed us to propose two models, showing the role of LmjMCA in the cell death and also in the autophagy pathway, implicating different protein domains.

Enzymes of yeast polyphosphate metabolism: structure, enzymology and biological roles.

Inorganic polyphosphate (polyP) is found in all living organisms. The known polyP functions in eukaryotes range from osmoregulation and virulence in parasitic protozoa to modulating blood coagulation, inflammation, bone mineralization and cellular signalling in mammals. However mechanisms of regulation and even the identity of involved proteins in many cases remain obscure. Most of the insights obtained so far stem from studies in the yeast Saccharomyces cerevisiae. Here, we provide a short overview of the properties and functions of known yeast polyP metabolism enzymes and discuss future directions for polyP research.

Review and new therapeutic alternatives for the treatment of cutaneous Leishmaniasis

Leishmaniasis comprises a group of diseases caused by protozoa of the genus Leishmania and has two basic clinical forms, visceral Leishmaniasis and cutaneous Leishmaniasis. The clinical features of Leishmaniasis depend on the species of Leishmania, the interaction between host and parasite and the immune response. This work focuses on cutaneous leishmaniosis because although it is not a deadly disease it results in significant scars and facial disfigurements, thus being clinically important. Furthermore, the first-line treatment consists of intravenous or intramuscular administration of intralesional pentavalent antimonials, which are highly toxic, making hospitalization of patients compulsory during treatment, with the associated financial costs. Herein, we review studies on drugs and treatments with fewer side effects and easier routes of administration such as topical administration. Recent research shows that the topical route of administration holds promise for the future treatment of cutaneous leishmaniosis.

Review and new therapeutic alternatives for the treatment of cutaneous Leishmaniasis

Leishmaniasis comprises a group of diseases caused by protozoa of the genus Leishmania and has two basic clinical forms, visceral Leishmaniasis and cutaneous Leishmaniasis. The clinical features of Leishmaniasis depend on the species of Leishmania, the interaction between host and parasite and the immune response. This work focuses on cutaneous leishmaniosis because although it is not a deadly disease it results in significant scars and facial disfigurements, thus being clinically important. Furthermore, the first-line treatment consists of intravenous or intramuscular administration of intralesional pentavalent antimonials, which are highly toxic, making hospitalization of patients compulsory during treatment, with the associated financial costs. Herein, we review studies on drugs and treatments with fewer side effects and easier routes of administration such as topical administration. Recent research shows that the topical route of administration holds promise for the future treatment of cutaneous leishmaniosis.

Compostagem de resíduos da indústria de desfibrilação de algodão

O processo de compostagem foi avaliado por meio do monitoramento diário da temperatura em leiras confeccionadas com o resíduo da indústria de desfibrilação de algodão e tipos de inóculo. Foram avaliadas seis leiras, divididas em dois sistemas: com aeração e sem aeração, sendo estudados, em cada sistema, três tipos de inóculo: esterco bovino diluído em água; esterco bovino, e conteúdo ruminal. Avaliou-se, também, a porcentagem de redução de volume nas leiras quando se intensificaram os revolvimentos, visando a obter informações para fins de dimensionamento do pátio de compostagem. Os compostos produzidos foram transferidos para minhocário e amostras do produto após a compostagem e após a vermicompostagem foram analisadas quimicamente com o objetivo de avaliar os teores de nutrientes. Os resultados obtidos demonstraram que o conteúdo ruminal utilizado como inóculo tende a apresentar valores mais altos de temperatura na fase inicial (fase ativa) e menores na fase final (maturação), embora, independentemente do tipo de inóculo utilizado, a estabilização praticamente tenha ocorrido no mesmo período; o sistema com aeração possibilitou, em todos os tratamentos, estabilização mais rápida do material, quando comparado ao sistema sem aeração; a intensificação dos revolvimentos possibilitou diminuição do tempo de retenção com redução de 46% no volume final, e a composição química dos vermicompostos apresentou-se enriquecida quando comparada aos compostos produzidos.