Periodic changes in chemical composition and in vitro digestibility of locally available Gramineae feed resources in the Philippines.

Efficient utilization of locally available feed resources is needed for further development of livestock productions in the tropics. However, an inadequate supply of nourish grass is common, and limited information exists regarding the chemical composition and digestibility of locally available feed resources in the different season. There were few reliable information concerning nutritive value of Gramineae resources in dry and rainy seasons in the Philippines. Hence, the present study was conducted to identify the chemical composition and in vitro digestibility of nine kinds of locally available Gramineae feed resources in dry and rainy seasons in the Philippines. The concentration of dry matter (DM), crude protein, and crude fiber of grass samples did not differ among species and seasons. The concentration of organic matter (OM), ether extract (EE), crude ash (CA), nitrogen-free extract (NFE), acid detergent fiber, and neutral detergent fiber (NDF) of grass samples differed significantly among species in both seasons except the NDF in the rainy season. However, the OM, CA, and NDF concentrations of the samples showed no differences between seasons. Imperata cylindrica showed the highest concentration of OM among the samples. Brachiaria brizantha had the lowest EE concentration among the grasses. The lowest NFE concentration was identified in Panicum maximum among the grasses. Although the digestibility of DM (DMD), OM (OMD), and NDF (NDFD) of grass samples showed significant differences among species. The DMD, OMD, and NDFD of I. cylindrica were the lowest among the samples, though Pennisetum purpureum showed the highest DMD, OMD, and NDFD among the species. Relatively high nutritive values of P. purpureum were remarkable among the samples in the present survey area.

Changes in nutrient composition and in vitro ruminal fermentation of total mixed ration silage stored at different temperatures and periods.

BACKGROUND: Total mixed ration (TMR) is widely used for dairy cattle and needs to be prepared daily because it deteriorates rapidly. Ensiling TMR allows preservation and saves labour at the farm; however, silage fermentation may influence various nutritional components. The objectives of this study were to evaluate nutritional changes and in vitro rumen fermentation of TMR silage that was stored at different temperatures and durations on a laboratory scale in comparison with those of typical TMR before ensiling. RESULTS: No distinct changes in crude protein (CP), neutral detergent fibre and non-fibrous carbohydrate contents were observed during silage fermentation. However, clear changes were observed in the soluble CP and soluble sugar fractions; solubilisation of the CP fraction in TMR silage was enhanced by prolonged storage and higher storage temperatures, and most soluble sugars were lost during ensiling. Short-chain fatty acid concentrations in the in vitro rumen from TMRs before and after ensiling were not significantly different; however, throughout incubation, NH3 -N concentrations from TMR silages were significantly higher than those from TMR before ensiling. CONCLUSION: A higher ruminal NH3 -N concentration from TMR silage may be a result of a shortage of fermentable sugars and enhanced deamination of CP. Feeding TMR ensiled under a high temperature must be investigated to balance proteins and carbohydrates for rumen fermentation.

Ruminal fermentation and microbial ecology of buffaloes and cattle fed the same diet.

Although buffaloes and cattle are ruminants, their digestive capabilities and rumen microbial compositions are considered to be different. The purpose of this study was to compare the rumen microbial ecology of crossbred water buffaloes and cattle that were fed the same diet. Cattle exhibited a higher fermentation rate than buffaloes. Methane production and methanogen density were lower in buffaloes. Phylogenetic analysis of Fibrobacter succinogenes-specific 16S ribosomal RNA gene clone library showed that the diversity of groups within a species was significantly different (P < 0.05) between buffalo and cattle and most of the clones were affiliated with group 2 of the species. Population densities of F.succinogenes, Ruminococcus albus and R. flavefaciens were higher until 6 h post-feeding in cattle; however, buffaloes exhibited different traits. The population of anaerobic fungi decreased at 3 h in cattle compared to buffaloes and was similar at 0 h and 6 h. The diversity profiles of bacteria and fungi were similar in the two species. The present study showed that the profiles of the fermentation process, microbial population and diversity were similar in crossbred water buffaloes and crossbred cattle.

Comparative analysis of methanogen diversity in the rumen of crossbred buffalo and cattle in the Philippines by using the functional gene mcrA.

Comparative analyses of methanogen diversity in the rumen of crossbred buffalo and cattle fed the same diet in the Philippines was performed by cloning the methyl coenzyme M reductase A (mcrA) gene. The cattle and buffalo libraries consisted of 50 clones each. Comparative analysis of the amino acid sequence revealed that these 2 libraries differed significantly (P < 0.01). The deduced amino acid sequences of the clones were classified into 9 operational taxonomic units (OTUs) in buffalo and 11 OTUs in cattle. Sequence similarity between the clones and known cultured methanogens ranged from 86 to 97 % for buffalo and 84 to 99 % for cattle. Methanobrevibacter species were predominant in buffalo (64 % of the clones), and an unknown mcrA was predominant in cattle (52 % of the clones). A large number of clones with low similarity to cultivated methanogens was observed in both buffalo and cattle, suggesting the presence of an unknown methanogen species in their rumen.

Metabolism of microbial nitrogen in ruminants with special reference to nucleic acids.

Characteristically the metabolism of microbial nitrogen (N) compounds in ruminants involves the degradation of dietary N and synthesis of microbial protein (MP), compounds including a small amount of peptides and free amino acids, which may account for 75-85% of total N and the remainder are nucleic acids (NA: DNA and RNA). Generally rumen microbes contain 10-25% NA-N of the total N while 70-80% is in the form of RNA. This paper describes the degradation and synthesis of NA in the rumen and their fate in the lower digestive tracts. Their physiological and nutritional significance in different types of ruminant animals is also discussed. The research works on NA metabolism in ruminants has been mainly on metabolism of purines after rumen microbial digestion and absorption in the lower gut. Subsequently, the fate of absorbed purines has been intensively investigated to assess the extent of MP synthesis in the rumen. The method for predicting ruminal synthesized MP and subsequently digested MP has been proposed using urinary purine derivative (PD) excretion in sheep and cattle fed on ordinary feed. The latter approach has now been adopted for calculation of protein supply in some feeding standards, although there are still difficulties in predicting representative samples of rumen microbes, and also uncertainties in variations of non-renal and endogenous purine losses.

Comparative study of plasma leptin concentration between solid ruminal and liquid abomasal feeding in weaned adult sheep.

This experiment was conducted to investigate the difference between ruminal (solid feed, SF) and abomasal (liquid feed, LF) feeding on the plasma leptin concentration in sheep. The experiment consisted of 2 weeks to adapt the animals to SF, 4 weeks of feeding on SF, 2 weeks adaptation to LF, 8 weeks of feeding on LF, 2 weeks of adaptation to SF, and 4 weeks of feeding on SF. The LF directory flowed into the abomasums of sheep by bottle feeding. Plasma leptin concentration before morning feeding was almost constant in the SF periods, whereas it showed between-day variations when measured during the LF periods. Mean plasma leptin levels were higher for LF (7.77 ± 0.76 ng/mL; mean ± SE) than for SF periods (3.95 ± 0.16 ng/mL; mean ± SE). Although plasma leptin concentration did not show any change after feeding in the SF and LF periods, plasma insulin and glucose levels increased within 15 min after liquid abomasal feeding, but not after solid ruminal feeding. The high plasma leptin concentration during the LF periods in weaned sheep could be due to change of digestible energy intake and changes in plasma insulin and glucose levels accompanying the changes in digestive processes and nutrient supply.

Eating and rumination behaviour in Brahman grade cattle and crossbred water buffalo fed on high roughage diet.

The purpose of this study is to compare feeding and other behavior and nutrient digestibility of tropical grade Brahman (body weight (BW) = 231 kg ± 12.4; n = 3) and crossbred water buffalo (BW = 300 kg ± 13.9; n = 3). This experiment on digestibility and measures of muscles of mastication utilized one-way, and animal behavior two-way, analysis of variance, respectively. Two video camcorders were installed in each pair of buffalo and Brahman for 24 h period programmed on the 107th, 109th and 111th days of the digestion trials. Frequency and duration of feeding, meal intake, rumination, bolus, chews, drinking, defecating, standing and lying were recorded daily. Muscle diameter of Digastricus, Masseter and Pterygoid and different regions of the tongue were sampled and measured under light microscope using a standard micrometer. Buffalo obtained significantly higher intake of dry matter, roughage, crude protein, total digestible nutrient and metabolized energy than Brahman. This was supported by longer meal duration (P ≤ 0.05), and shorter meal breaks (P ≤ 0.05) of buffalo than Brahman. The diameter of the muscles for mastication was bigger (P ≤ 0.05) in buffalo than in Brahman, which is indicative of stronger chewing ability. Briefly, lesser and slower chewing action; higher intake of roughage and crude protein; and longer resting behavior of crossbred water buffalo than Brahman are all indicative of better digestive and metabolic performance of the buffalo under high roughage feeding conditions.

Effect of flow parameters of rumen digesta on effective degradability and microbial yield in sheep.

The aim of this experiment was to examine the effect of rumen digesta flow parameters on effective degradability (EDG) and microbial nitrogen (MBN) yield in sheep fed diets of identical provision of both metabolizable energy and rumen degradable nitrogen (RDN). Ruminal degradation parameters of early-harvested perennial ryegrass hay (EH), late-harvested perennial ryegrasses hay (LH) and winter sown barley straw (BS) were determined by a nylon bag technique. Subsequently, three experimental diets (EHD, LHD and BSD) were formulated using the tested forages, sucrose and urea as supplements. An in vivo feeding study was conducted using four rumen cannulated sheep in a partial Latin square design. Digestibility, rate constants of rumen particle breakdown and passage, ruminal fermentation parameters and MBN supply to the small intestine were determined. Animals thoroughly consumed the forages fed at a restricted level. Digestion coefficients were greater for EHD than for LHD and were lowest for BSD (P < 0.05). The rate constant of large particle breakdown was 4.3, 5.9 and 6.7 %/h, respectively, and small particle passage was 5.3, 4.7 and 6.3 %/h for EHD, LHD and BSD, respectively. The estimates differed (P < 0.05) between the diets. The overall passage rate constant of total rumen particles was estimated to be higher for BSD than that for EHD or LHD (P < 0.05). Ruminal fermentation parameters were unaffected by dietary treatments (P > 0.05). Intake levels of rumen degradable organic matter (RDOM) and RDN were estimated to be greater for LHD than that for EHD or BSD (P < 0.05). Although dietary arrangements were made to give identical microbial efficiency, the estimated value was higher for EHD than that for LHD or BSD (P < 0.05); and the MBN yield for BSD was estimated to be lower than that for EHD or LHD (P < 0.05). Rumen kinetic parameters of degradation and particle flow of forage affected EDG values and MBN yield from forage-related RDN intake, although those had little effect on the efficiency of MBN yield from forage-related RDOM intake.

The content of protein, fibre and minerals of leaves of selected Acacia species indigenous to north-western Tanzania.

Browse tree leaves of six species of Acacia (A. angustissima L., A. drepanolobium L., A. nilotica L., A. polyacantha L., A. senegal L., A. tortilis L.) were screened for chemical composition, including minerals and trace elements. Crude protein (CP) varied among the species from 145 (A. senegal) to 229 g/kg DM (A. angustissima). The species had moderate to high levels of minerals. The concentrations of Ca, P, Mg and S varied among the species from 14.6-31.5, 3.5-4.9, 1.4-3.0 and 1.7-2.8 g/kg DM, respectively. The forages showed relatively low concentrations of trace elements. Content of trace elements varied among the species from 4.5-23.8, 99.4-173.6, 146.2-432, 41.0-90.1, 10.9-22.2 and 0.05-0.65 mg/kg DM for Cu, Mo, Fe, Mn, Zn and Co, respectively. All leaves of browse species would meet the normal requirements for Ca, P, Mg and S in ruminants, although some species had higher levels of Ca than tabulated mineral requirements in livestock. Assayed Cu, Mn, Zn and Co would satisfy the lower range of recommended requirements of trace elements depending on their bioavailability. Therefore, browse leaves from Acacias could form good sources of CP and mineral supplements to ruminants.

Which genetic marker for which conservation genetics issue?

Conservation genetics focuses on the effects of contemporary genetic structuring on long-term survival of a species. It helps wildlife managers protect biodiversity by identifying a series of conservation units, which include species, evolutionarily significant units (ESUs), management units (MUs), action units (AUs), and family nets (FNs). Although mitochondrial DNA (mtDNA) evolves 5-10 times faster than single-copy nuclear DNA (scnDNA), it records few traces of contemporary events. Thus, mtDNA can be used to resolve taxonomic uncertainties and ESUs. Variable number of tandem repeats (VNTRs) evolve 100-1000 times faster than scnDNA and provide a powerful tool for analyzing recent and contemporary events. VNTR analysis techniques include polymerase chain reaction (PCR)-based microsatellite assays and oligonucleotide probing. Size homoplasy problems in PCR-based microsatellite assays can strongly affect the inference of recent population history. The high homozygosity in endangered species is reflected in a relatively low number and level of variability in microsatellite loci. This combined with "allelic dropout" and "misprinting" errors contributes to the generation of highly biased genetic data following analyses of natural populations. Thus, in conservation genetics, microsatellites are of limited use for identifying ESUs, MUs, and AUs. In contrast to PCR-based microsatellite analysis, oligonucleotide probing avoids errors resulting from PCR amplification. It is particularly suitable for inferring recent population history and contemporary gene flow between fragmented subpopulations. Oligonucleotide fingerprinting generates individual-specific DNA banding patterns and thus provides a highly precise tool for monitoring demography of natural populations. Hence, DNA fingerprinting is powerful for distinguishing ESUs, MUs, AUs, and FNs. The use of oligonucleotide fingerprinting and fecal DNA is opening new areas for conservation genetics.

Diquafosol elicits increases in net Cl- transport through P2Y2 receptor stimulation in rabbit conjunctiva.

The purpose of the present study was to understand the mechanisms of action of diquafosol, a stable derivative of uridine 5'-triphosphate, on Cl(-) transport across the isolated rabbit conjunctiva. Rabbit conjunctivas were isolated and mounted in a modified Ussing chamber. Under short-circuit conditions, the effects were determined of mucosal (tear) side diquafosol application on the short-circuit current (Isc). Diquafosol rapidly and dose-dependently increased the Isc at concentrations ranging from 0.1 to 968 microM when added to the mucosal side of the conjunctiva. In the absence of the serosal Cl(-), the Isc induced by 10 microM diquafosol was substantially reduced. On the contrary, in the absence of mucosal side Na(+), the diquafosol-induced increases in Isc were unchanged. Following 45-min preincubation, the P2Y(2) antagonist suramin inhibited the diquafosol-induced increases in the Isc whereas the P2Y(1) antagonist pyridoxal-phosphate-6-azophenyl-2'4'-disulfonic acid had no effect. These studies suggest that diquafosol stimulates net Cl(-) secretion from the serosal to the mucosal side via stimulation of P2Y(2) receptors in the rabbit conjunctiva.

Durability of FTLA treatment as a medicament for dentin hypersensitivity--abrasion resistance and profiles of fluoride release.

The purpose of this study was to evaluate the durability of tubules occluded with FTLA treatment by toothbrush abrasion test on the applied surface and by measuring fluoride release from the FTLA components. Dentin specimens with simulated hypersensitive surfaces were treated with APF containing tannic acid. After which, the specimens received lanthanum-chloride-with-powdered-fluoroapatite-glass-ceramics treatment. The specimens were subjected to toothbrush abrasion test up to 6,000 strokes. SEM observation revealed that dentinal tubules of the FTLA treated specimens were completely occluded with fine deposits even after toothbrush abrasion of 6,000 strokes. EPMA analysis revealed that fluoride, lanthanum, and aluminum were the main FTLA components on the dentin surface after 6,000-stroke abrasion. To measure fluoride release from the FTLA components, a slurry was enclosed in a cellulose tube and suspended in deionized water at 37 degrees C. After fluoride was dialyzed against deionized water, a high concentration of fluoride was found to be released from FTLA the components, indicating FTLA treatment's prominent durability. These results suggested that FTLA treatment has a superior resistance against toothbrush abrasion and a high fluoride-releasing performance. These characteristics lend much weight to showing that the FTLA method is an effective and durable medicament for dentin hypersensitivity.

P2Y(2) receptor elicits PAS-positive glycoprotein secretion from rabbit conjunctival goblet cells in vivo.

We investigated in vivo whether UTP and ATP increased periodic acid and Schiff's reagent (PAS)-positive glycoprotein release from rabbit conjunctival goblet cells. Fifty microL of UTP or ATP at the concentrations of 0.003, 0.03, 0.3, 3.0, 8.5% (54 microM-154 mM) or saline were applied to rabbit eyes. Impression cytology was performed on the upper nasal bulbar conjunctiva and the cells were stained with PAS. To clarify purinergic receptor-mediated involvement in this response, suramin (1%; 7 mM), P2Y(2) antagonist and pyridoxal-phosphate-6-azophenyl-2',4'-disulphonic acid (PPADS, 0.01%; 167 microM), P2Y(1) antagonist were applied in the rabbit conjunctival sac for 10 min. before UTP or ATP application. Images of the specimens were taken with a digital camera mounted on a microscope and the PAS staining area was measured using an image analyzing system. UTP or ATP eye drop instillation transiently decreased the PAS staining area in a dose-dependent manner, but it gradually recovered after another 30 min. Saline instillation had no effect until 60 min. later. All of the agonists-induced declines were inhibited by pretreatment with 1% (7 mM) suramin but not 0.01% (167 microM) PPADS. UTP and ATP stimulate PAS-positive glycoprotein secretion via P2Y(2) receptor on goblet cells in the rabbit bulbar conjunctiva in vivo.

Genetic differentiation and subspecies development of the giant panda as revealed by DNA fingerprinting.

Over the last 100 years giant pandas (Ailuropoda melanoleuca) have been separated into six completely isolated mountain ranges. DNA fingerprinting revealed different differentiation patterns in giant pandas, including early-stage, late-stage, recent divergence and recent genetic depression. A separation around 10 000 years ago resulted in highly significant differences in DNA fingerprints and morphological characteristics between Qinling and Sichuan populations. Supported by morphological differences, the genetic data were used to classify the Qinling population as a new subspecies, A. m. qinlingensis, while the Sichuan populations were classified into the original subspecies, A. m. melanoleuca. Thus, the Qinling population deserves management as a separate unit. In the Sichuan populations, two management units were defined, including Qionglai-Minshan and Daxiangling-Xiaoxiangling-Liangshan. Our data suggest urgent measures are needed to establish green corridors between subpopulations in each mountain range to increase gene flow and genetic variation to ensure long-term survival.

Novel noninvasive sensitive determination of tear volume changes in normal cats.

We describe a novel, high-resolution and noninvasive method for measuring tear volume changes in cats. The method entails photographing at the lid margin the tear meniscus area defined by instillation of 0.1% fluorescein solution into the cul-de-sac. The inferior tear meniscus area was obtained from the digitized images with computer-assisted software. The tear meniscus area increased in proportion to the saline volume applied into the conjunctival sac, which validates the technique. Furthermore, this technique detected with high sensitivity previously described increases in tear fluid secretion induced by the P2Y(2) agonist. We demonstrate in cats that changes in conjunctival sac tear volume can be evaluated by measurement of its inferior tear meniscus area.

INS365 suppresses loss of corneal epithelial integrity by secretion of mucin-like glycoprotein in a rabbit short-term dry eye model.

P2Y2 receptor agonists, like UTP and ATP, stimulate mucin secretion from goblet cells in vitro. Therefore, mucin stimulants could be good candidates for the treatment of dry eye syndrome because mucin increases the tear film stability and protects against desiccation of ocular surface. INS365 is a more stable P2Y2 receptor agonist than UTP. In the present study, we evaluated, in normal rabbit eyes, its effectiveness to release mucin from goblet cells and to protect the corneal damage induced by desiccation. For mucin secretion, impression cytology was performed following the instillation of INS365 solution or saline into the conjunctival sac. The specimens were stained with periodic acid and Schiff (PAS) reagent, and then the staining area was calculated using computer software. INS365 dose-dependently decreased the PAS staining area of conjunctival goblet cells from 2 to 15 min post-application. Furthermore, we utilized the rabbit short-term dry eye model to evaluate if INS365 eyedrops could protect against any of the damage produced by blockage of blinking with ocular speculum. INS365 significantly suppressed corneal damage at concentrations of more than 0.1% w/v. These results suggest that this P2Y2 agonist is a good candidate for the treatment of dry eye disease.