Maternal rumen and milk microbiota shape the establishment of early-life rumen microbiota in grazing yak calves.

Early-life gut microbial colonization and development exert a profound impact on the health and metabolism of the host throughout the life span. The transmission of microbes from the mother to the offspring affects the succession and establishment of the early-life rumen microbiome in newborns, but the contributions of different maternal sites to the rumen microbial establishment remain unclear. In the present study, samples from different dam sites (namely, oral, rumen fluid, milk, and teat skin) and rumen fluid of yak calves were collected at 6 time points between d 7 and 180 postpartum to determine the contributions of the different maternal sites to the establishment of the bacterial and archaeal communities in the rumen during early life. Our analysis demonstrated that the dam's microbial communities clustered according to the sites, and the calves' rumen microbiota resembled that of the dam consistently regardless of fluctuations at d 7 and 14. The dam's rumen microbiota was the major source of the calves' rumen bacteria (7.9%) and archaea (49.7%) compared with the other sites, whereas the potential sources of the calf rumen microbiota from other sites varied according to the age. The contribution of dam's rumen bacteria increased with age from 0.36% at d 7 to 14.8% at d 180, whereas the contribution of the milk microbiota showed the opposite trend, with its contribution reduced from 2.7% at d 7 to 0.2% at d 180. Maternal oral archaea were the main sources of the calves' rumen archaea at d 14 (50.4%), but maternal rumen archaea became the main source gradually and reached 66.2% at d 180. These findings demonstrated the potential microbial transfer from the dam to the offspring that could influence the rumen microbiota colonization and establishment in yak calves raised under grazing regimens, providing the basis for future microbiota manipulation strategies during their early life.

Small intestinal morphology and sugar transporters expression when consuming diets of different energy levels: comparison between Tibetan and small-tailed Han sheep.

Some non-structural carbohydrates, especially starch, escape ruminal fermentation, are converted into glucose, and are absorbed from the small intestine. This glucose provides an important source of energy, and its usage is more efficient than glucose from carbohydrates which are fermented as short chain fatty acids in the rumen and, subsequently, undergo hepatic gluconeogenesis. Tibetan sheep graze on the harsh Qinghai-Tibetan Plateau (QTP) all year round and their carbohydrate and energy intakes fluctuate greatly with seasonal forage availability. Consequently, a high capacity to absorb glucose from the small intestine would be particularly beneficial for Tibetan sheep to allow them to cope with the inconsistent dietary intakes. This study examined how the small intestinal morphology and sugar transporters' expression of Tibetan and Small-tailed Han (Han) sheep respond to fluctuating energy intakes under the harsh conditions of the QTP. Han sheep graze on the QTP only in summer and are generally raised in feedlots. Twenty-four Tibetan sheep and 24 Han sheep, all wethers, were assigned randomly to four groups (n = 6 per breed/group), with each group offered a diet differing in digestible energy content: 8.21, 9.33, 10.45 and 11.57 MJ/kg DM. After 49 d, all sheep were slaughtered, tissues of the small intestine were collected, and measurements were made of the morphology and glucose transporters and the related regulation gene expressions. At intakes of low energy levels, Tibetan sheep had a greater villus surface area in the duodenum, jejunum and ileum and higher mRNA expression of sodium-dependent glucose transporter 1 in the duodenum and ileum (P < 0.05) than Han sheep. In the glucose transporter 2 (GLUT2) mediated glucose absorption pathway, Tibetan sheep had higher GLUT2 and taste receptor family 1 member 2 and 3 mRNA expressions than Han sheep in the duodenum, jejunum and ileum (P < 0.05). We concluded that the differences between breeds indicated a greater glucose absorption capacity in the small intestine of Tibetan than Han sheep, which would confer an advantage to Tibetan over Han sheep to an inconsistent energy intake on the harsh QTP. These findings suggested that ruminants raised under harsh environmental conditions with highly fluctuating dietary intakes, as is often the case in grazing ruminants worldwide, are able to absorb glucose from the small intestine to a greater extent than ruminants raised under more moderate conditions.

Energy substrate metabolism in skeletal muscle and liver when consuming diets of different energy levels: comparison between Tibetan and Small-tailed Han sheep.

The energy intake of Tibetan sheep on the harsh Qinghai-Tibetan Plateau (QTP) varies greatly with seasonal forage fluctuations and is often below maintenance requirements, especially during the long, cold winter. The liver plays a crucial role in gluconeogenesis and skeletal muscle is the primary tissue of energy expenditure in mammals. Both play important roles in energy substrate metabolism and regulating energy metabolism homeostasis of the body. This study aimed to gain insight into how skeletal muscle and liver of Tibetan sheep regulate energy substrate metabolism to cope with low energy intake under the harsh environment of the QTP. Tibetan sheep (n = 24; 48.5 ± 1.89 kg BW) were compared with Small-tailed Han sheep (n = 24; 49.2 ± 2.21 kg BW), which were allocated randomly into one of four groups that differed in dietary digestible energy densities: 8.21, 9.33, 10.45 and 11.57 MJ /kg DM. The sheep were slaughtered after a 49-d feeding period, skeletal muscle and liver tissues were collected and measurements were made of the activities of the key enzymes of energy substrate metabolism and the expressions of genes related to energy homeostasis regulation. Compared with Small-tailed Han sheep, Tibetan sheep exhibited higher capacities of propionate to glucose conversion and fatty acid oxidation and ketogenesis in the liver, higher glucose utilization efficiency in both skeletal muscle and liver, but lower activities of fatty acid oxidation and protein mobilization in skeletal muscle, especially when in negative energy balance. However, the Small-tailed Han sheep exhibited higher capacities to convert amino acids and lactate to glucose and higher levels of glycolysis and lipogenesis in the liver than Tibetan sheep. These differences in gluconeogenesis and energy substrate metabolism conferred the Tibetan sheep an advantage over Small-tailed Han sheep to cope with low energy intake and regulate whole-body energy homeostasis under the harsh environment of the QTP.

Comparison of nitrogen utilization and urea kinetics between yaks (Bos grunniens) and indigenous cattle (Bos taurus).

Under traditional management on the Qinghai-Tibetan Plateau, yaks () graze only on natural pasture without supplements and are forced to cope with sparse forage of low N content, especially in winter. In contrast, indigenous Tibetan yellow cattle () require supplements during the cold season. We hypothesized that, in response to harsh conditions, yaks cope with low N intakes better than cattle. To test this hypothesis, a study of whole-body N retention and urea kinetics was conducted in 2 concurrent 4 × 4 Latin squares, with 1 square using yaks and 1 square using cattle. Four isocaloric forage-concentrate diets differing in N concentrations (10.3, 19.5, 28.5, and 37.6 g N/kg DM) were formulated, and by design, DMI were similar between species and across diets. Urea kinetics were determined with continuous intravenous infusion of NN urea for 104 h, and total urine and feces were concomitantly collected. Urea production, urea recycling to the gut, and ruminal microbial protein synthesis all linearly increased ( < 0.001) with increasing dietary N in both yaks and cattle. Urinary N excretion was less ( = 0.04) and N retention was greater ( = 0.01) in yaks than in cattle. Urea production was greater in yaks than in cattle at the 3 lowest N diets but greater in cattle than in yaks at the highest N diet (species × diet, < 0.02). Urea N recycled to the gut ( < 0.001), recycled urea N captured by ruminal bacteria ( < 0.001), and ruminal microbial protein production ( = 0.05) were greater in yaks than in cattle. No more than 12% of urea recycling was through saliva, with no difference between species ( = 0.61). Glomerular filtration rate was lower ( = 0.05) in yaks than in cattle. The higher urea recycling and greater capture of recycled urea by ruminal microbes in yaks than in cattle suggest that yaks use mechanisms to utilize dietary N more efficiently than cattle, which may partially explain the better survival of yaks than cattle when fed low-N diets.

Protein and mRNA expression of follicle-stimulating hormone receptor and luteinizing hormone receptor during the oestrus in the yak (Bos grunniens).

Follicle-stimulating hormone (FSH) and luteinizing hormone (LH) have a central role in follicle growth, maturation and oestrus, but no clear pathway in the seasonal oestrus of yak (Bos grunniens) has been found. To better understand the role of FSH and LH in seasonal oestrus in the yak, six yaks were slaughtered while in oestrus, and the pineal gland, hypothalamus, pituitary gland, and gonads were collected. Using real-time PCR and immunohistochemical assays, we determined the mRNA and protein expression of the FSH and LH receptors (FSHR and LHR) in these organs. The analysis showed that the FSHR mRNA expression level was higher in the pituitary gland tissue compared with LHR (p < .01) during oestrus. By contrast, there was low expression of FSHR and LHR mRNA in the pineal gland and hypothalamus. FSHR mRNA expression was higher than that of LHR (p < .05) in the ovary, whereas LHR mRNA expression was higher than that of FSHR (p < .01) in the uterus. FSHR and LHR proteins were located in the pinealocyte, synaptic ribbon and synaptic spherules of the pineal gland and that FSH and LH interact via nerve fibres. In the hypothalamus, FSHR and LHR proteins were located in the magnocellular neurons and parvocellular neurons. FSHR and LHR proteins were localized in acidophilic cells and basophilic cells in the pituitary gland, and in surface epithelium, stromal cell and gland epithelium in the uterus. In the ovary, FSHR and LHR protein were present in the ovarian follicle. Thus, we concluded that FSHR and LHR are located in the pineal gland, hypothalamus, pituitary and gonad during oestrus in the yak. However, FSHR was mainly expressed in the pituitary gland and ovaries, whereas LHR was mainly expressed in the pituitary gland and uterus.

Cloning of oligopeptide transport carrier PepT1 and comparative analysis of PepT1 messenger ribonucleic acid expression in response to dietary nitrogen levels in yak () and indigenous cattle () on the Qinghai-Tibetan plateau.

The gastrointestinal lumen can directly absorb all di- and tripeptide protein degradation products, and oligopeptide absorption depends on the specific peptide transport carriers, which are located in gastrointestinal epithelial cells on the brush border membrane. Yak () use N more efficiently than cattle do, which implies that yak have a specific mechanism of nonprotein utilization including a peptide absorption mechanism. However, this mechanism has not been clarified. Our objective was to explore whether yak possess any adaptive mechanisms of peptide absorption to survive in the harsh foraging environment of the Qinghai-Tibetan plateau. Twelve castrated males of each of 2 genotypes, yak () and indigenous cattle (), were fed diets of various N levels. The yak PepT1 (yPepT1) cDNA was cloned in omasum epithelial tissue. Our results showed that the full-length yPepT1 cDNA contains 2,805 bp, and a 2,121-bp open reading frame encodes a putative protein of 707 AA residues. The yPepT1 AA sequence identified 5 putative extracellular N-glycosylation sites (Asn, Asn, Asn, Asn, and Asn), 2 putative intracellular protein kinase A sites (Ser and Thr), and 3 intracellular putative protein kinase C sites (Ser, Ser, and Ser). The yPepT1 AA sequence was 99, 95, 86, and 83% identical to PepT1 from cattle (), sheep (), pigs (), and humans (), respectively. The relative PepT1 mRNA expression for indigenous cattle was greater than yak in the rumen, omasum, duodenum, ileum, and liver ( < 0.001); however, it was lower in jejunum tissue ( < 0.01). The relative PepT1 mRNA expression in response to increasing dietary N for both genotypes were linear in the rumen and jejunum ( < 0.10); quadratic or cubic in the reticulum ( < 0.01); linear or quadratic in the duodenum, ileum, and liver ( ≤ 0.01); and linear, quadratic, or cubic in the omasum ( < 0.001). Moreover, there were significant interactions between genotype and dietary N in rumen, reticulum, omasum, duodenum, jejunum, ileum, and liver tissues. In conclusion, the PepT1 profile and expression in gastrointestinal epithelial cells of yak varied from those of cattle, implying that yak have evolved a peptide transport mechanism to adapt the environment of the Qinghai-Tibetan plateau.

Isolation, identification and fibrolytic characteristics of rumen fungi grown with indigenous methanogen from yaks (Bos grunniens) grazing on the Qinghai-Tibetan Plateau.

AIM: To obtain co-cultures of anaerobic fungi and their indigenously associated methanogens from the rumen of yaks grazing on the Qinghai-Tibetan Plateau and investigate their morphology features and ability to degrade lignocellulose. METHODS AND RESULTS: Twenty fungus-methanogen co-cultures were obtained by Hungate roll-tube technique. The fungi were identified as Orpinomyces, Neocallimastix and Piromyces genera based on the morphological characteristics and internal transcribed spacer 1 sequences analysis. All methanogens were identified as Methanobrevibacter sp. by 16S rRNA gene sequencing. There were four types of co-cultures: Neocallimastix with Methanobrevibacter ruminantium, Orpinomyces with M. ruminantium, Orpinomyces with Methanobrevibacter millerae and Piromyces with M. ruminantium among 20 co-cultures. In vitro studies with wheat straw as substrate showed that the Neocallimastix with M. ruminantium co-cultures and Piromyces with M. ruminantium co-cultures exhibited higher xylanase, filter paper cellulase (FPase), ferulic acid esterase, acetyl esterase activities, in vitro dry matter digestibility, gas, CH4 , acetate production, ferulic acid and p-coumaric acid releases. The Neocallimastix frontalis Yak16 with M. ruminantium co-culture presented the strongest lignocellulose degradation ability among 20 co-cultures. CONCLUSIONS: Twenty fungus-methanogen co-cultures were obtained from the rumen of grazing yaks. The N. frontalis with M. ruminantium co-cultures were highly effective combination for developing a fermentative system that bioconverts lignocellulose to high activity fibre-degrading enzyme, CH4 and acetate. SIGNIFICANCE AND IMPACT OF THE STUDY: The N. frontalis with M. ruminantium co-cultures from yaks grazing on the Qinghai-Tibetan Plateau present great potential in lignocellulose biodegradation industry.

Short communication: Changes in the composition of yak colostrum during the first week of lactation.

Although the great interest has been paid to colostrum utilization for calves, no systematic studies evaluating the compositional changes of yak colostrum during the first week after parturition have been reported. The aim of this study was to elucidate such postpartum nutritional changes. Colostrum samples from 12 multiparous (2-7 lactations) yaks, grazed on alpine pasture, were collected at exactly 1, 24, 48, 72, 96, 120, 144, and 168 h postpartum. Gross composition (fat, total solids, protein, and ash) were measured, as well as fat and water-soluble vitamins, fatty acid and mineral composition, and IgG. The colostrum, collected 1h postpartum, had the highest concentrations of crude protein, total solids, ash, vitamin A and E, ß-carotene, and most minerals (Na, Mg, Zn, Cu, and Fe). These components decreased rapidly within 24h. Similarly, at 24h postpartum, IgG content decreased to 7.5% of the value (87.78 mg/mL) at 1h postpartum. In contrast, the concentration of vitamin C increased from 1 to 24h and then decreased consistently thereafter to the lowest value at 168 h. Phosphorus and Ca contents showed an increasing trend from 24 to 168 h after calving. Lactose content increased from 2.88% at 1h to 4.96% at 48 h postpartum and was steady to 168 h. Total n-3 fatty acids, monounsaturated fatty acids, polyunsaturated fatty acids, and n-3-to-n-6 fatty acid ratio were proportionally similar as a percentage of total fatty acid methyl esters during the first 168 h of milk production. However, the proportion of total n-6 fatty acid content to total fatty acid methyl esters decreased from 3.07% at 1h to 2.60% at 24h. In summary, experimental results indicate colostrum should be provided to yak calves as soon as possible after birth to provide nutrients and enhance passive immunity.

A comparison of nitrogen utilization and urea metabolism between Tibetan and fine-wool sheep.

To study metabolic adaptation to harsh foraging conditions, an experiment was conducted to characterize and quantify N utilization efficiency and urea metabolism in Tibetan and fine-wool sheep fed 4 levels of dietary N (11.0, 16.7, 23.1, and 29.2 g N/kg DM) in 2 concurrent 4 × 4 Latin square designs. Urea kinetics were determined using continuous intrajugular infusions of 15N15N-urea. Urinary excretions of total N and urea N increased linearly (P < 0.001) with dietary N and were not different between breeds (P ≥ 0.37). Fecal N excretion increased with dietary N for Tibetan sheep but not for fine-wool sheep (linear dietary N × breed; P < 0.05). Nitrogen retention (both amount per day and percentage of N intake) increased with increasing dietary N concentration (P < 0.001), and the rates of increase were greater in fine-wool than in Tibetan sheep (linear dietary N × breed and cubic dietary N × breed; P < 0.05). In Tibetan sheep, N retention as a percentage of intake was greatest for diets containing 16.7 g N/kg DM, whereas it was maximal for fine-wool sheep when the diet contained 23.1 g N/kg DM. Urea N entry rate, urea N recycled to the gastrointestinal tract (GIT), and urea N returned to the ornithine cycle all increased with dietary N (P < 0.05), and all were greater in Tibetan than fine-wool sheep for the 11.0 g N/kg DM diet but were greater in fine-wool than Tibetan sheep for the diet with 29.2 g N/kg DM (linear dietary N × breed; P < 0.05). Urea N excreted in feces, both amount and fraction of GIT entry rate, was less in Tibetan than fine-wool sheep for the 11.0 and 16.7 g N/kg DM diets but similar for diets with 23.1 or 29.2 g N/kg DM (linear dietary N × breed; P < 0.01). For the lowest-protein diet, the fraction of urea N production recycled to the GIT was greater in the Tibetan than fine-wool sheep (88% vs. 82%), but for the diet with 29.2 g N/kg DM it was greater for fine-wool than Tibetan sheep (46% vs. 39%; linear dietary N × breed; P < 0.05). Plasma urea N increased more rapidly in response to increasing dietary N concentration for fine-wool sheep than for Tibetan sheep (linear dietary N × breed; P < 0.05). Urea tubular load and the amount and percentage of urea reabsorbed by the kidney were greater in Tibetan than fine-wool sheep (P < 0.05). These results suggest that Tibetan sheep have mechanisms that allow them to utilize N more efficiently than the fine-wool sheep when dietary N is inadequate.

Grasshopper (Orthoptera: Acrididae) community composition in the rangeland of the northern slopes of The Qilian Mountains in northwestern China.

In order to describe grasshopper (Orthoptera: Acrididae) species composition, diversity, abundance, and density of four rangelands types, we compared the grasshopper community composition and dynamics in the rangeland of the northern slopes of the Qilian Mountains. In total, 55 grasshopper species were collected from 2007 to 2009, representing three families and six subfamilies. The subfamily Oedipodinae was dominant, followed by Gomphocerinae and Catantopinae. Species abundance varied among rangeland types (RTs). The greatest abundance of grasshoppers was found in mountain rangeland, while the lowest abundance of grasshoppers was caught in alpine shrublands. Three species (Chorthippus cf. brunneus (Thunberg) (Acrididae), Chorthippus Dubius (Zubovski), and Gomphocerus licenti (Chang) were broadly distributed in the four RTs and constituted 7.5% of all grasshoppers collected. Ch. dubius was very abundant in desert rangeland and alpine shrubland. Bryodema dolichoptera Yin et Feng Eremippus qilianshanensis Lian and Zheng, and Filchnerella qilianshanensis Xi and Zheng (Pamphagidae) were endemic to the region of the Qilian Mountains. Species similarity between RTs ranged from 17.8 to 51.6 based on the Renkonen index. Similarly, the Sörensen index indicated a wide separation in species composition among RTs. The abundance of the eight most common species showed obvious differences among RTs and years. On average, mountain rangeland had the highest density values in 2007 and 2008, and alpine shrubland supported the smallest density. The densities in desert and mountain rangeland in 2007 were significantly higher than in 2008, while alpine rangeland and shrublands did not present obvious differences among years.

The effect of a diet containing grasshoppers and access to free-range on carcase and meat physicochemical and sensory characteristics in broilers.

1. Research was conducted to evaluate the impact of a diet containing grasshoppers on the carcase, physicochemical and sensory characteristics in a free-range, grassland-based broiler production system. 2. A total of 80, 28-d-old male broilers were reared on grassland containing a large population of grasshoppers (treatment PB). Control birds were reared intensively on a maize-soybean diet (treatment CB). At 91 d of age, 24 birds from each treatment were slaughtered to evaluate carcase, meat and sensory characteristics. 3. Treatment PB produced birds with significantly lower live weights, breast, wing, thigh and drum weights, and higher dressing percentage and breast percentage of carcase, compared with CB. Treatment PB produced breast meat with significantly higher redness values, shear force and protein content, and lower pH values, cooking loss, moisture and fat content compared with CB. Sensory panel results for breast and thigh meats showed no treatment effect on colour and juiciness, but significantly higher scores for chewiness, flavour, aroma and overall appreciation, and lower scores for tenderness from treatment PB compared with CB. 4. Rearing chickens on rangeland may provide an alternative way to produce poultry meat which is considered superior by modern consumers.

Effects of plant enzyme inactivation or sterilization on lipolysis and proteolysis in alfalfa silage.

This experiment studied the contribution of plant enzymes and microbial activity on lipolysis and proteolysis in ensiled alfalfa. Before ensiling, the wilted alfalfa was treated with plant enzyme inactivation by autoclaving or with sterilization by γ-ray irradiation. The treated alfalfa was then inoculated with commercial lactic acid bacteria inoculants and ensiled for 40 d. Alfalfa without treatment was ensiled as the control. The content of total fatty acid (FA) after ensiling decreased 43% in the control silage and 28% in the γ-ray-treated silage, but did not change in the autoclave-treated silage. Among the major FA (C16:0, C18:2n-6, C18:3n-3), a considerable increase was observed in proportion of C16:0 in the control silage as compared with fresh alfalfa; conversely, decreases in proportions of C18:2n-6 and C18:3n-3 occurred during ensilage. Silage treated with γ-ray radiation at ensiling had a smaller proportion of C16:0 and greater proportions of C18:2n-6 and C18:3n-3 than control silage. Autoclave treatment further decreased proportions of C16:0 and most of the other FA, and increased C18:2n-6 and C18:3n-3 proportions in comparison with γ-ray treatment. Proportions of C16:0, C18:2n-6, C18:3n-3 and other detected FA (except for the proportion of C15:0) did not differ between fresh forage and autoclave-treated silage. Remarkably, smaller nonprotein nitrogen content was observed in the autoclave-treated silage compared with the γ ray-treated silage or the control silage. These results indicated that an extensive lipolysis occurred during ensiling of alfalfa, and plant enzymes played a major role in lipolysis and proteolysis.

Aspects of lipid oxidation of meat from free-range broilers consuming a diet containing grasshoppers on alpine steppe of the Tibetan Plateau.

The objective of this study was to evaluate the characteristics of lipid oxidation in the meat of free-range broilers feeding on grasshoppers in alpine rangeland on the Tibetan Plateau. Eighty 28-d-old Qinjiaoma male broilers were introduced into a rangeland where there was a dense population of grasshoppers (PB). Control birds were reared under intensive conditions and given a maize-soybean diet. At 91 d of age, 24 birds from each treatment were slaughtered. Fresh breast and thigh meats were packaged and refrigerated for determination. The results indicate that rearing conditions, which included a diet rich in grasshoppers for PB broilers, significantly (P < 0.05) affected α-tocopherol content, total iron, heme iron, and nonheme iron content in the muscle of both the breast and leg. Rearing system and diet also had an important effect on antioxidant activity and lipid oxidation during refrigerated storage. This influence changed with storage time between different tissues. The activities of glutathione peroxidase and superoxide dismutase in the muscle of chicken breasts or legs from PB broilers were significantly (P < 0.05) higher than those from the controls, but no significant (P > 0.05) differences were found for the activities of catalase between the PB broilers and the controls. In conclusion, the meat in free-range broilers feeding on grasshoppers has more antioxidative potential and longer storage life.

Contribution of exopeptidases to formation of nonprotein nitrogen during ensiling of alfalfa.

The experiment was conducted to investigate the exopeptidase classes in alfalfa (Medicago sativa L.) leaves, and to determine their contribution to the formation of nonprotein nitrogen (NPN) components during ensiling. Six classes of inhibitors that included bestatin (aminopeptidase inhibitor), potato carboxypeptidase inhibitor (PCI, carboxypeptidase inhibitor), 1,10-phenanthroline (dipeptidase inhibitor), diprotin A (dipeptidyl-peptidase inhibitor), butabindide (tripeptidyl-peptidase inhibitor), and dipeptide Phe-Arg (peptidyl-dipeptidase inhibitor) were used. To determine the contribution of each exopeptidase to the formation of NPN products, aqueous extracts of fresh alfalfa were fermented to imitate the proteolytic process of ensiled alfalfa and to ensure that each class of exopeptidase inhibitor would have immediate contact with the proteases in the alfalfa extract. Five classes of exopeptidases; namely, aminopeptidase, carboxypeptidase, dipeptidase, dipeptidyl-peptidase, and tripeptidyl-peptidase, were shown to be present in alfalfa leaves, each playing a different role in alfalfa protein degradation. Aminopeptidase, carboxypeptidase, and dipeptidase were the main exopeptidases contributing to the formation of NH(3)-N. Among the 5 exopeptidases, tripeptidyl-peptidase appeared to be the principal exopeptidase in hydrolyzing forage protein into peptides, whereas carboxypeptidase and dipeptidase appeared to be more important in contributing to the formation of amino acid-N. Dipeptidyl-peptidase and tripeptidyl-peptidase did not play a role in the formation of NH(3)-N or amino acid-N. Dipeptidase, carboxypeptidase, and tripeptidyl-peptidase were the principal exopeptidases for hydrolyzing forage protein into NPN during ensilage, and treatment with a mixture of the 5 inhibitors reduced the total NPN concentration in the fermented alfalfa extract to about 45% of that in the control after 21 d of fermentation.

Effect of strategic feed supplementation on productive and reproductive performance in yak cows.

A total of 2230 yak cows (5-13 years of age) in two populations with different milking systems were investigated. One population had a system of milking once a day (MOD), and the other population twice a day (MTD). The average milk yield of MOD cows was 0.7 +/- 0.2 kg/day within a milking period of 109 +/- 9 days. This compared with an average of 1.24 +/- 0.3 kg/day in 127 +/- 6 days in MTD yaks (p < 0.01). The cows showed a calving rate of 71% under the MOD system and 51.4% under the MTD system. Three farms with a total of 104 MTD cows between six and 12 years of age were used to provide three different feeding groups. The groups were fed with, or without oat hay or highland barley straw in amounts of 1-1.5 kg/head/day from December to April. The three farms were designated as Farm I, Farm II, and Farm III. Farm I had 41 cows with body weight of 230 +/- 67 kg each for grazing with no supplement (GNS). Farm II had 30 animals with body weight of 216 +/- 28 kg each for grazing + oat hay (GOH). Farm III had 33 animals body weight of 221 +/- 34 kg each for grazing + highland barely straw (GBS). The calving rates of the cows in GOH and GBS were 23 and 19% higher, respectively, than GNS cows (p < 0.01), and the highest rate reached 76.7% in GOH. The live weight loss of the cows in GNS was considerably higher (p < 0.01) than in the two other groups. Ten GOH cows and 12 GNS cows were used to collect milk samples for measuring the progesterone concentration using RIA kits provided by IAEA/FAO: Milk was sampled every five days from calving until 90 days postpartum. In the unsupplemented group, milk progesterone (P4) levels suggested that cows had started cyclic ovarian activity by 40 days postpartum, whereas only 25% had been observed in estrus. In the supplemented group, 80% of cows had started cyclic ovarian activity by the same time and 70% had been seen in estrus. Two types of cyclic activity in terms of progesterone changed were found. With Type I (normal), 50 and 80% of cows from GNS and GOH, respectively, had cyclic changes of P4 in milk at 40 days postpartum. With Type II, the P4 levels in the milk remained 0.89 ng/ml until 90 days postpartum. A total of 46 grazing cows between five and 13 years of age (body weight 214 +/- 68 kg) was used to collect blood samples to measure concentrations of nutrient of metabolites at two weeks pre-calving and at two weeks, two months and four months postcalving, respectively. The concentrations of nutrient metabolites [albumin, globulin, urea, beta-hydroxybutyrate (BHB) and inorganic phosphorus] suggested general underfeeding of energy and protein in the winter/late pregnancy period with some recovery in lactation. Energy constraints appeared again as the summer progressed. No dietary phosphate deficiency was found. BHB and albumin testing on serum yaks could be a useful tool to identify poor nutritional status during the winter and so illustrate the need for supplementation.

Occupational injuries: a study of health care workers at a northwestern health science center and teaching hospital.

1. This retrospective study examined the magnitude of the problem of work related health hazards affecting professional and non-professional employees of a state university health science center and hospital. 2. From the 1,513 injury episodes that occurred among the 9,668 employees during the 32 month study period, it was determined that the highest risk categories for both department and job were nursing, housekeeping, food service, and laboratory technicians. 3. Injury rate for females (11.2 per 100 person years) was more than twice that of males (5.1). Injury rates declined from 11.6 per 100 person years at ages 25 to 39 to 3.8 at ages over 60. Puncture wounds (32.7%), mostly by needlesticks, were the most frequently reported injury type, followed by strains and sprains (17.2%), lacerations (12.5%), and contusions (12.1%).

Inability of awake patients to correctly locate a cough stimulus.

To determine whether patients can accurately identify the lung in which a cough reflex originates, a unilateral cough stimulus was administered to 67 patients during fiberoptic bronchoscopy. Although the stimulus produced a vigorous cough reflex in all subjects, 51 of 67 individuals were unable to identify the side stimulated. These results suggest that impulses from endobronchial irritant receptors are not perceived cortically as originating within a particular hemithorax.