Sources of error with in vitro digestibility assay of pasture feeds

A series of experiments was undertaken to determine population statistics for in vitro organic matter digestibility ( in vitro OMD) data and to examine the effects of basal diet, donor animal and precollection fasting interval on the activity and specificity of rumen fluid inoculum. The experiments utilized wether sheep, a diverse set of pasture grass and legume feeds prominent in the Australian subtropics and the Tilley and Terry in vitro digestibility procedure running under the operating pressure of a practicing feeds evaluation laboratory.
The standard errors of in vitro OMD estimates for within and between batch runs were ±0·88 × 10⁻² and 0·62 × 10⁻², respectively. These error terms were used to develop protocols to accept, reject or scale raw in vitro OMD data. Differences between donor animals in the activity of rumen fluid were highly significant. Extending the precollection fasting interval beyond 16 h was associated with a substantial decline in inoculum activity.
An in vitro-in vivo calibration relationship based on fifteen test feeds and using lucerne ( Medicago sativa ) as basal diet was described by the linear model y = 1·3 x-0·195±4·9 × 10⁻² r = 0·79 (y = in vivo OMD, x = in vitro OMD). Despite large effects of basal diet on both the absolute values and relative ranking of test feeds, neither the RSD nor r values were improved using alternative diets to Lucerne chaff.
The results highlight the need to formally standardize the analytical and biological components of the in vitro digestibility procedure to safeguard the integrity of data.     

Effects of monensin on Mg, Ca, P and Zn metabolism and tissue concentrations in lambs

A study consisting of two trials was conducted to determine the effects of monensin on the apparent absorption and retention of magnesium (Mg), phosphorus (P), calcium (Ca) and zinc (Zn) and to determine mineral changes in tissue and ruminal fluid. Eight lambs (39 kg) were used in trial 1, and 10 lambs (37 kg) were used in trial 2. Animals were blocked by weight and fed a high concentrate diet with or without 20 mg/kg monensin. Trials began with a dietary adjustment period lasting 18 d in trial 1 and 21 d in trial 2. Animals were then placed in metabolism stalls for a 10-d stall adjustment period followed by a 12-d collection period. Collections to determine mineral balance were made during the first 10 d of the collection period. Blood and ruminal fluid samples were taken on d 11 of the collection period. Lambs were slaughtered on d 12 of the collection period and tissue samples were collected. Monensin supplementation increased (P less than .05) Mg retention 42.0%. Urinary Ca excretion decreased (P less than .05) 60.0% when monensin was fed. Monensin supplementation decreased (P less than .05) liver Ca and bone Ca, 45.5 and 2.9%, respectively. Apparent P digestibility increased (P less than .05) 40.0% and P retention increased (P less than .10) 26.8% due to monensin supplementation. Both apparent absorption and retention of Zn increased (P less than .01) 50.0 and 45.0%, respectively, with monensin supplementation. Ruminal fluid Zn concentrations decreased (P less than .05) 33.0% with the addition of monensin.(ABSTRACT TRUNCATED AT 250 WORDS)     

Identification of Cytokeratins in Bovine Sperm Outer Dense Fibre Fractions

Outer dense fibres (ODF) are important substructures of mammalian sperm tails that are involved in the regulation of sperm motility. In this study, we investigated the identity of several sodium dodecyl sulphate (SDS)‐insoluble ODF proteins. Bovine ODF were purified by separating sperm heads and tails using ultrasound and Percoll^® density gradient centrifugation. Sperm flagella were treated with the detergent cetyltrimethylammonium bromide (CTAB). CTAB‐insoluble material, which reportedly represents the ODF fraction, was collected, and electron microscopy confirmed a highly purified ODF fraction. We found after solubilization of this fraction with SDS that high amounts of insoluble material were retained after centrifugation. SDS‐insoluble material was collected and quantitatively dissolved in 8 M urea. SDS‐gel electrophoresis in the presence of urea revealed polypeptides with apparent molecular masses of approximately 25, 43, and 50 kDa. Subsequent immunoblotting with anti‐cytokeratin antibodies detected two urea‐soluble, SDS‐insoluble proteins with apparent molecular masses of 45 and 66 kDa. The 45‐kDa protein was identified as cytokeratin 19. An antibody reacting with a palette of cytokeratins (CK 1–18 and CK 20), KL1, was the only antibody that reacted with the 66‐kDa polypeptide. We conclude that sperm ODF fractions contain at least one each of type I and type II intermediate filaments. As keratins and intermediate filaments are described as rope‐like structures, we suggest that these intermediate filaments play an important structural or tension‐bearing role in sperm flagella.