Avaliação da ocorrência de Giardia spp. por diferentes métodos coprológicos

O género Giardia inclui espécies com potencial zoonótico e com uma distribuição mundial, e em que alguns genótipos de Giardia duodenalis são responsáveis anualmente por milhares de novos casos em humanos. Existem vários ciclos de transmissão, sendo a água de consumo, por contaminação fecal de origem animal, uma das principais fontes de infecção humana. Neste estudo foram colhidas 162 amostras fecais de animais de quatro origens diferentes (Zoológico = 55; Produção = 25; Doméstico = 5; Canil = 77) que foram testadas por duas técnicas coprológicas diferentes, a técnica de flutuação com sacarose e a técnica de sedimentação com formol-acetato. Para a implementação de Nested PCR foram testados vários genes, β-giardina, Glutamato desidrogenase e 18SrRNA, ocorrendo apenas amplificação das amostras com o gene 18SrRNA. Com esta técnica foram analisadas 26 amostras, que incluía a treze positivas à microscopia e as restantes escolhidas aleatoriamente. Este trabalho permitiu determinar a ocorrência de Giardia spp. através das técnicas coprológicas em treze animais de diferentes origens e verificar que o número de animais de canil positivos não foi o esperadas de acordo com o descrito na literatura que refere ser este o grupo com maior prevalência. Este estudo também permitiu uma comparação entre os dois métodos de concentração de quistos de Giardia spp., com maior recuperação utilizando a técnica de flutuação com sacarose. Através da técnica molecular confirmaram-se dez dos positivos encontrados por microscopia e ainda se detectaram dois novos positivos.

Nitrosospira spp. can produce nitrous oxide via a nitrifier denitrification pathway

Nitrous oxide (N2O) emission from soils is a major contributor to the atmospheric loading of this potent greenhouse gas. It is thought that autotrophic ammonia oxidizing bacteria (AOB) are a significant source of soil-derived N2O and a denitrification pathway (i.e. reduction of NO2- to NO and N2O), so-called nitrifier denitrification, has been demonstrated as a N2O production mechanism in Nitrosomonas europaea. It is thought that Nitrosospira spp. are the dominant AOB in soil, but little information is available on their ability to produce N2O or on the existence of a nitrifier denitrification pathway in this lineage. This study aims to characterize N2O production and nitrifier denitrification in seven strains of AOB representative of clusters 0, 2 and 3 in the cultured Nitrosospira lineage. Nitrosomonas europaea ATCC 19718 and ATCC 25978 were analysed for comparison. The aerobically incubated test strains produced significant (P < 0.001) amounts of N2O and total N2O production rates ranged from 2.0 amol cell(-1) h(-1), in Nitrosospira tenuis strain NV12, to 58.0 amol cell(-1) h(-1), in N. europaea ATCC 19718. Nitrosomonas europaea ATCC 19718 was atypical in that it produced four times more N2O than the next highest producing strain. All AOB tested were able to carry out nitrifier denitrification under aerobic conditions, as determined by production of N-15-N2O from applied N-15-NO2-. Up to 13.5% of the N2O produced was derived from the exogenously applied N-15-NO2-. The results suggest that nitrifier denitrification could be a universal trait in the betaproteobacterial AOB and its potential ecological significance is discussed.

Genotypic differences in the growth of bananas (Musa spp.) infected with migratory endoparasitic nematodes. 1. Roots

The effects of nematodes on root morphology and the association of root characteristics with resistance to nematodes of seven banana varieties were investigated in two experiments. Banana plants were grown in controlled conditions within polytunnels and harvested on three occasions for the measurement of root morpholopy, and biomass. Varieties differed in their resistance to nematodes, from resistant (Yg Km5, FHIA 17, FHIA 03) and partly resistant (FHIA 01, FHIA 25) to not resistant ((FHIA 23, Williams). Nematodes reduced the root dry weight of FHIA 01, FHIA 17 and FHIA 23 at some harvests. Primary root number was on average 9.5% lower in nematode-infected plants than controls, with no differences among the varieties. Thus, there was no simple association between the resistance of these varieties and their tolerance to nematodes. Varieties differed in root morphology. Root dry weight was greatest for resistant varieties Yg Km5 and FHIA 03, and least for non-resistant varieties FHIA 23 and Williams. Thus, resistance to nematodes was associated with varieties with greater root mass and more and larger primary roots.

Genotypic differences in the growth of bananas (Musa spp.) infected with migratory endoparasitic nematodes. 2. Shoots

In order to identify the effect of burrowing nematodes on the shoots (pseudostem and leaves) of banana plants and to determine whether or not shoot characteristics are associated with plant resistance to nematodes two experiments were conducted in controlled conditions within polytunnels. The banana plants were harvested on three occasions for the measurement of root morphology and biomass. Varieties differed in their resistance to nematodes from resistant (Yg Km5, FHIA 17, FHIA 03) and partly resistant (FHIA 01, FHIA 25) to not resistant (FHIA 23, Williams). Nematodes reduced total plant dry weight at the first harvest in Experiment 1 and by an average of 8.8% in Experiment 2, but did not affect leaf area in either experiment. The ratio of above-ground Weight to total plant weight was reduced from 75% to 72% in nematode-infected plants compared with the control plants for all varieties tested in Experiment 1, but was only reduced in FHIA 25 and FHIA 23 in Experiment 2. Varieties differed in above-ground growth. The FHIA varieties had greater shoot weights and leaf area than YgKm5 and Williams. Overall, resistance to nematodes was associated with the partitioning of a greater proportion of biomass to the roots than to above-ground parts.

Involvement of phenazine-1-carboxylic acid, siderophores and hydrogen cyanide in suppression of Rhizoetonia solani and Pythium spp. damping-off by Pseudomonas oryzihabitans and Xenorhabdus nematophila

Entomopathogenic bacterial strains Pseudomonas (Flavimonas) oryzihabitans and Xenorhabdus nematophilus, both bacterial symbionts of the entomopathogenic nematodes Steinernema abbasi and S. carpocapsae have been recently used for suppression of soil-borne pathogens. Bacterial biocontrol agents (P. oryzihabitans and X nematophila) have been tested for production of secondary metabolites in vitro and their fungistatic effect,on mycelium and spore development of soil-borne pathogens. Isolates of Pythium spp. and Rhizoctonia solani, the causal agent of cotton damping-off, varied in sensitivity in vitro to the antibiotics phenazine-I-carboxylic acid (PCA), cyanide (HCN) and siderophores produced by bacterial strains shown previously to have potential for biological control of those pathogens. These findings affirm the role of the antibiotics PCA, HCN and siderophores in the biocontrol activity of these entomopathogenic strains and support earlier evidence that mechanisms of secondary metabolites are responsible for suppression of damping-off diseases. In the present studies colonies of R oryzihabitans showed production of PCA with presence of crystalline deposits after six days development and positive production where found as well in the siderophore's assay when X nematophila strain indicated HCN production in the in vitro assays. In vitro antifungal activity showed that bacteria densities of 101 to 10(6)cells/ml have antifungal activity in different media cultures. The results show further that isolates of Pythium spp. and R. solani insensitive to PCA, HCN and siderophores are present in the pathogen population and provide additional justification for the use of mixtures of entomopathogenic strains that employ different mechanisms of pathogen suppression to manage damping-off.

Prevalence and determinants of Cryptosporidium spp. infection in smallholder dairy cattle in Iringa and Tanga Regions of Tanzania

The prevalence of Cryptosporidium spp. infection in a cross-sectional study of dairy cattle, from two contrasting dairying regions in Tanzania, were determined by staining smears of faecal samples with the modified Ziehl-Neelsen technique. Of the 1126 faecal samples screened, 19.7% were positive for Cr\yptosporidium spp. The prevalence was lower in Tanga Region than in Iringa Region. The prevalence of affected farms was 20% in Tanga and 21 % in Iringa. In both regions, the probability of detecting Cryptosporidium oocysts in faeces varied with animal class, but these were not consistent in both regions. In Tanga Region, Cryptosporidium oocysts were significantly more likely to be found in the faeces of milking cows. In Iringa Region, the likelihood that cattle had Cryptosporidium-positive faeces declined with age, and milking cattle were significantly less likely to have Cryptosporidium-positive faeces. In this region, 7% of cattle were housed within the family house at night, and this was marginally associated with a higher likelihood that animals had Ctyptosporidium-positive faeces. Our study suggests that even though herd sizes are small, Cryptosporidium spp. are endemic on many Tanzanian smallholder dairy farms. These protozoa may impact on animal health and production, but also on human health, given the close associations between the cattle and their keepers. Further studies are required to assess these risks in more detail, and understand the epidemiology of Cryptosporidium spp. in this management system.

Farm-level risk factors for fluoroquinolone resistance in E-coli and thermophilic Campylobacter spp. on finisher pig farms

Logistic regression, supported by other statistical analyses was used to explore the possible association of risk factors with the fluoroquinolone (FQ)-resistance status of 108 pig finisher farms in Great Britain. The farms were classified as 'affected' or 'not affected' by FQ-resistant E. coli or Campylobacter spp. on the basis of isolation of organisms from faecal samples on media containing 1 mg/l FQ. The use of FQ was the most important factor associated with finding resistant E. coli and/or Campylobacter, which were found on 79% (FQ-resistant E. coli) and 86% (FQ-resistant Campylobacter) of farms with a history of FQ use. However, resistant bacteria were also found on 19% (FQ-resistant E. coli) and 54% (FQ-resistant Campylobacter) of farms with no history of FQ use. For FQ-resistant E. coli, biosecurity measures may be protective and there was strong seasonal variation, with more farms found affected when sampled in the summer. For FQ-resistant Campylobacter, the buying-in of grower stock may increase risk and good on-farm hygiene may be protective. The findings suggest that resistant organisms, particularly Campylobacter, may spread between pig farms.