Mohammad Rezaeian

Distribution and estimation of anaerobic zoosporic fungi along the digestive tracts of sheep.

The status of anaerobic zoosporic (Chytridiomycota) fungi along the entire digestive tract of sheep was assessed both analytically and microscopically. Digest samples were taken from different segments of the digestive tracts of three newly killed sheep that previously had been used in experimental dietary studies. These digest samples were tested for the presence of rumen fungi by assessing the recovery of live fungi from the samples, direct observation of digested plant remains under the scanning electron microscope (SEM), and using a chitin assay as an estimation of fungal biomass. Live anaerobic fungi were recovered from the abomasum, small and large intestine, caecum and faeces of sheep, but not from the digest samples of rumen and omasum. However, SEM examination of the samples confirmed the presence of fungal structures from all of these organs. In the large intestine and caecum samples the observed sporangial structures were rounded and showed conspicuous surface pitting. Results of the chitin assay indicated that the anaerobic fungi might account for up to 20% of the total microbial biomass in the rumen of sheep. The results of this study support the view that anaerobic fungi may be present as a resistant stage in the lower reaches of the digestive tract.

Methods for the isolation, culture and assessment of the status of anaerobic rumen chytrids in both in vitro and in vivo systems

Anaerobic fungi were isolated from both the rumen and faeces of nine sheep and a cow. A reliable and simple method for the isolation of anaerobic fungi using 24 h rumen incubated milled straw as the inoculum source was developed. We also evaluate the use of chitin measurements as an assay of rumen fungal biomass. Chitin levels were determined from various sample sources (milled barley straw used as the fungal culture substrate in vitro; plant particulate digests from the rumen (PLP) and centrifuged strained rumen fluid (CSRF) using both HPLC and colorimetric methods. Both methods were highly correlated and consequently the simpler colorimetric method was adopted for subsequent studies. There was also a high degree of correlation between anaerobic fungal cellulase activities with the assayed chitin content of milled barley straw cultures over 12 d of an in vitro experiment. The colorimetric chitin assay protocol was then used to assess the diurnal variation and abundance of rumen fungi in in vivo assays. We assessed the distribution of chitin (mg g(-1) dry matter) in various fractions of the strained rumen fluid (SRF) and PLP samples from the rumen of sheep. Chitin was detected in all fractions of strained rumen fluid but the main source of chitin in the samples may be attributed to the fungal biomass. We did not detect any significant differences in chitin levels over a 24 h sampling period. Finally, an SEM study on subsamples of milled straw and plant particulate matter used in the chitin assays, revealed that the pattern of the fungal development on substrate material differs from the culture medium to the rumen.

Simultaneous optimization of power and duration of radio-frequency pulse in PARACEST MRI.

Chemical exchange saturation transfer (CEST) MRI is increasingly used to probe mobile proteins and microenvironment properties, and shows great promise for tumor and stroke diagnosis. The CEST effect is complex and depends not only on the CEST agent concentration, exchange rates, the characteristic of the magnetization transfer (MT), and the relaxation properties of the tissue, but also varies with the experimental conditions such as radio-frequency (RF) pulse power and duration. The RF pulse is one of the most important factors that promote the CEST effect for biological properties such as pH, temperature and protein content, especially for contrast agents with intermediate to fast exchange rates. The CEST effect is susceptible to the RF duration and power. The present study aims at determining the optimal power and the corresponding optimal duration (that maximize the CEST effect) using an off-resonance scheme through a new definition of the CEST effect. This definition is formulated by solving the Bloch-McConnell equation through the R1ρ method (based on the eigenspace solution) for both of the MT and CEST effects as well as their interactions. The proposed formulations of the optimal RF pulse power and duration are the first formulations in which the MT effect is considered. The extracted optimal RF pulse duration and power are compared with those of the MTR asymmetry model in two- and three-pool systems, using synthetic data that are similar to the muscle tissue. To validate them further, the formulations are compared with the empirical formulation of the CEST effect and other findings of the previous researches. By extending our formulations, the optimal power and the corresponding optimal duration (in the biological systems with many chemical exchange sites) can be determined.

Effects of diets containing alfalfa hay or barley flour mixed alfalfa silage on feeding behavior, productivity, rumen fermentation and blood metabolites in lactating cows.

The effects of barley flour on the fermentation parameters of alfalfa silage and on the productivity of dairy cows were investigated. Alfalfa forage was ensiled either with or without barley flour. Barley flour was soaked in water for 24 h before being mixed with alfalfa (12 kg: 100 kg dry matter bases) at ensiling. Eighteen multi-parous cows were assigned to three equal treatment groups using a completely randomized design. Three isocaloric and isonitrogenous total mixed rations containing alfalfa hay, ordinary alfalfa silage or barley flour mixed alfalfa silage were then prepared. The concentrations of ammonia nitrogen, acetic acid and butyric acid were lower in barley flour mixed alfalfa silage compared to that in ordinary alfalfa silage but the concentration of lactic acid was lower in the ordinary alfalfa silage. Feeding behavior, milk yield and composition, ruminal fermentation and blood metabolites were measured. Although dry matter intake and milk production were not affected, the effect of preparation of alfalfa influenced feeding behavior and rumen fermentation parameters. Cows on alfalfa silage diets spent longer ruminating compared to those fed alfalfa hay. The ruminal ammonia nitrogen and blood urea were affected by ensiling (alfalfa hay versus alfalfa silages) while both parameters were lower in cows fed on barley flour mixed alfalfa silage than those fed on ordinary silage. Although similar blood glucose was recorded for cows fed on alfalfa silages, it was higher in cows fed on alfalfa hay. It is concluded that the addition of barely flour when making alfalfa silage may improve both the fermentation process during ensilage and the ruminal ammonia nitrogen utilization with no significant effects on productivity.