Nitrogen Metabolim in Sheep Fed Hay-Based Diet Supplemented with Yucca schidigera with or without Nisin

Empat ekor domba yang berfistula pada bagian rumen digunakan pada rancangan Bujur sangkar latin 3 x 3. penelitian ini bertujuan untuk mengetahui pengaruh suplementasi Yucca schidigera dengan atau tanpa nisin terhadap penggunaan nitrogen, konsentrasi amonia dalam rumen, konsentrasi nitrogen urea dalam plasma, suplai nitrogen mikroba pada domba yang diberi pakan basal hay rumput timothy dan konsentrat. Tiga perlakuan pakan yaitu pakan basal (kontrol), pakan basal + Yucca schidigera 240 ppm (yucca), pakan basal + Yucca schidigera 240 ppm + nisin 6 mg/kg BW 0.75 (yucca+nisin). Ekskresi nitrogen dalam urin domba yang diberi perlakuan yucca+nisin lebih rendah (P<0.05) dibandingkan perlakuan kontrol. Perlakuan yucca+nisin meningkatkan keseimbangan nitrogen sebesar 8 % dibandingkan kontrol. Konsentrasi amonia  dalam rumen pada perlakuan yucca dan yucca+nisin lebih rendah (P<0.05) daripada perlakuan kontrol, dan konsentrasi tersebut masih berada diatas konsentrasi minimal untuk pertumbuhan mikroba dalam rumen. Konsentrasi nitrogen urea dalam plasma tidak dipengaruhi oleh perlakuan dan bervariasi antara 20.8 – 22.0 mg/dl. Suplai nitrogen mikroba pada perlakuan yucca dan yucca+nisin lebih tinggi (P<0.05) dibandingkan perlakuan kontrol. Berdasarkan hasil penelitian ini dapat disimpulkan bahwa suplementasi yucca atau kombinasi yucca dan nisin dapat menurunkan konsentrasi amonia dalam rumen, sehingga menurunkan ekskresi nitrogen pada urin dimana hal ini berimplikasi positif dalam mengurangi pencemaran nitrogen di lingkungan. (Animal Production 7(1): 34-39 (2005) Kata kunci : Yucca schidigera, Nisin, Penggunaan N, N Mikroba

Influence of Non-fibrous Carbohydrate and Degradable Intake protein and Ruminal Fermentation ,Nutrien Digestion and performance of Local Sheep

The objective of current study was to evaluate the impact dietary non-fibrous carbohydrate ( NFC) and ruminally degradable intake protein (DIP) concentration have on ruminal fermentation , nutrient digestion and performance of local sheep. The animal had a mean of  liveweight 19.80 ±1.55 kg. four diets ,arranged in a 2x2 factorial ,were formulated to contain either 40 or 50 % NFC and 50 or 60 % of dietary crude protein as DIP .dietary DM contained 25 % Indonesian field grass and 75 % concentrate. Solvent –extracted or formaldehyd  2 % -treated soybean meal were used to alter DIP and corn or soybean hulls to alter NFC level. Percentage of  energy and NDF digestion was similar ( p<0,01) as DIP level decreased in the diets. The soybean hulls was fermentable and total VFA concentration in the rumen increased ( p<0.01), but N-NH3 concentration was decreased ( p<0.01) as DIP level decreased in the diets. Daily live weight gain ( 146.29±25.84 g) and body composition ( fat, water , protein and mineral) was similar ( p<0.05) among diets. The preponderance ruminal fermentation ,nutrient digestion and performance of local sheeps did not be improved by sincronization of energy and nitrogen release but may more likely be limited by either energy or nitrogen alone. (Animal Production 3(2): 53-61 (2001)Key Word : Carbohydrate, protein, rumen fermentation, nutrients digestion and performance

Voluntary Feed Intake and Growth of Local Sheep Fed on Road Side Grasses Following A 6-Week Feed Restriction Period

The purpose of this experiment was to investigate an adaptation time sheep to eat at a normal amount required by the local experiencing liveweight loss due to feed restriction, to investigate feed intake and feed digestibility and to investigate sheep growth following feed restriction. The experiment was conducted in The Experimental Farm of Faculty of Animal Husbandry, Unsoed. Twelve local sheep were randomly assigned to two different groups of treatment: namely, control (C) and restrictively Fed Animal (R). there were two periods through out the experiment, Period I (PI) and II (PII). During P I (6 week), Animals in group R were restrictively fed so that they lost live weight of approximately 15%. During P II Animals in group  R were than offered ad libitum. Whereas, animal in group C were fed ad libitum through out the experiment. The results of this experiment showed that animals in group R needed 17 days for adaptation to consume feed at a normal amount after they had experienced feed restriction for 6 weeks. Dry matter intake was not different between group C dan R. However, when it was expressed per metabolic live weight (g/Kg0,75), Dry matter intake for group R was significantly higher than that for group C (102.35 vs 91,79). Dry matter digestibility value for group R was higher than that for group C either during PI (62 vs 57%) or during PII (70 vs 61%). The rate of live weight gain was not different between C and R during period II. In conclusion, local sheep had capability of consuming more feed after resumption of full feeding for 17 days. (Animal Production 2(2): 47-52 (2000) Key words : local sheep, dry matter, organic matter, period, restriction, digestibility

The Effect of Urea and Sulphur Level in Mixing of Cassava Waste Fermented and Soybean Cake waste on Nitrogen Balance and Digestibility of Energy of Local Male Sheep

Cassava waste and soybean cake waste are by - product from home industry  that  have been used as animal feed. It contain high crude fibber, there fore it must be processed or added with another nutrients before feeding to animal to get a good performance. A research have been  conducted in Experimental Farm and Laboratory Animal of  Feed and Nutrition, Animal Science Faculty, Jenderal Soedirman University, Purwokerto at October- Nopember 2001. The purpose of  the research is to find the effect of urea and sulfur  level in mixing of cassava waste fermented and soybean cake waste on nitrogen balance and digestibility of energy of local male sheep. Eighteen local male sheep were  divided into 3 group based of the body weight. There are I = 18.55±1.27 kg, II = 15.79±0.67 kg and  III = 13.41±1.33 kg. Two level of urea (2% and 3%) and three level of sulfur (0 %, 0.15 % and 0.30%) dry matter concentrate used as  treatments, there fore factorial   design 2x3 with Randomized Block Design used this experiment All of treatment received same diet consist land grass and concentrates with 70:30 dry matter ratio. Concentrates consists  cassava waste fermented  and soybean cake waste with 77.50:22.50 dry matter ratio.  Dry matter intake was 4 % body weight. Observed variables were nitrogen balance and digestibility of energy by using total collection. Variance analysis was used for knowing the effect treatment on the variable observed. The result of the research shown that level of urea and sulfur are not significant interaction on the nitrogen balance and digestibility of energy.  However  level urea and sulfur  itself are significant on nitrogen balance linearly. Level urea also had significant effect on the energy digestion.  Balance nitrogen of all treatment is positive. Level of urea 3 % and sulfur 0.30% gave a good balance nitrogen and energy digestion.  It can be concluded that addition of urea and sulfur in diet containing fermentable  carbohydrates can increase balance nitrogen (N retention) and energy digestion. (Animal Production 3(2): 91-97 (2001)Key Words: Balance nitrogen, energy digestion, cassava waste and soybean cake waste

The Effect Of Feed Restriction And Exercise On Growt Of Merino Sheep

Penelitian dengan judul “Pengaruh Pembatasan  Pakan dan Exercise terhadap Pertumbuhan Domba Merino” dilakukan untuk mempelajari perubahan bobot badan dan komposisi tubuh selama periode pembatasan pakan dan kerja (exercise) yang kemudian diikuti oleh pemberian pakan secara adlibitum dan berhenti bekerja.penelitian ini terdiri dari dua periode.Pada periode pertama, 1,5 ekor Domba Merino kastrasi umur 4-5 bulan dibagi menjadi tiga perlakuan ,yaitu pembatasan pakan dan exercise selama 2,50 jam (perlakuan I), pembatasan pakan tanpa Exercise (perlakuan II), serta pemberian pakan secara ad libitum tanpa Exercise (perlakuan III), yang masing-masing perlakuan diulang lima kali. Pada periode kedua ,semua domba diberi pakan ad libitum tanpa exercise. Hasil penelitian menunjukankan bahwa pada akhir penelitian periode pertama ,domba dalam perlakuan satu I dan perlakuan II mengalami penurunan bobot badan sebesar 28 dan 27 persen dari bobot badan awal. Selama periode kedua, domba dalam perlakuan I dan perlakuan II tumbuh lebih cepat dari pada domba perlakuan III. Pada akhir periode pertama, domba dalam perlakuan I kehilangan lemak lebih banyak dari pada perlakuan II.Selama periode ke dua ,penimbunan protein pada perlakuan I dan II lebih cepat dibanding  perlakuan III. Domba pada perlakuan III mengalami penimbunan lemak lebih banyak dari pada perlakuan I dan II. Domba pada perlakuan I dan II selama periode ke dua mempunyai kemampuan makan lebih tinggi per kilogram bobot badan metabolis dibanding domba pada perlakuan III. (Animal Production 2(1): 18-24 (2000)Kata kunci: pembatasan pakan , komposisi tubuh , kemampuan makan, pertumbuhan.

Transition into high school : a phenomenological study

Transition is a process of moving from the known to the unknown (Green, 1997) and transition from primary school to high school can be described in this way. In Australia, there is a two tiered system of primary and secondary schooling operating where school students typically undergo at least two transitions. Firstly, when they leave home to attend pre-school/primary school; and secondly, when they leave primary school to enter secondary school. Potentially some students may experience up to four transitions: from home to kindergarten to pre-school to primary school to secondary school while for a smaller number of students who attend P-12 schools, there may be only one transition: from home to preschool.

Effect of surgical approach on bone vascularisation, fracture and soft tissue healing : comparison of less invasive to open approach

Over the past ten years, minimally invasive plate osteosynthesis (MIPO) for the fixation of long bone fractures has become a clinically accepted method with good outcomes, when compared to the conventional open surgical approach (open reduction internal fixation, ORIF). However, while MIPO offers some advantages over ORIF, it also has some significant drawbacks, such as a more demanding surgical technique and increased radiation exposure. No clinical or experimental study to date has shown a difference between the healing outcomes in fractures treated with the two surgical approaches. Therefore, a novel, standardised severe trauma model in sheep has been developed and validated in this project to examine the effect of the two surgical approaches on soft tissue and fracture healing. Twenty four sheep were subjected to severe soft tissue damage and a complex distal femur fracture. The fractures were initially stabilised with an external fixator. After five days of soft tissue recovery, internal fixation with a plate was applied, randomised to either MIPO or ORIF. Within the first fourteen days, the soft tissue damage was monitored locally with a compartment pressure sensor and systemically by blood tests. The fracture progress was assessed fortnightly by x-rays. The sheep were sacrificed in two groups after four and eight weeks, and CT scans and mechanical testing performed. Soft tissue monitoring showed significantly higher postoperative Creatine Kinase and Lactate Dehydrogenase values in the ORIF group compared to MIPO. After four weeks, the torsional stiffness was significantly higher in the MIPO group (p=0.018) compared to the ORIF group. The torsional strength also showed increased values for the MIPO technique (p=0.11). The measured total mineralised callus volumes were slightly higher in the ORIF group. However, a newly developed morphological callus bridging score showed significantly higher values for the MIPO technique (p=0.007), with a high correlation to the mechanical properties (R2=0.79). After eight weeks, the same trends continued, but without statistical significance. In summary, this clinically relevant study, using the newly developed severe trauma model in sheep, clearly demonstrates that the minimally invasive technique minimises additional soft tissue damage and improves fracture healing in the early stage compared to the open surgical approach method.

Agglutinins from marine macroalgae of the southeastern United States

Protein extracts from 22 species of marine macroalgae from Florida and North Carolina were compared for their abilities to agglutinate sheep and rabbit erythrocytes. Protein extracts from 21 algal species agglutinated rabbit erythrocytes compared to 19 for sheep erythrocytes. However, agglutination by brown algal extracts was variable. The agglutination produced by protein extracts from Dictyota dichotoma could be blocked by addition of polyvinylpyrrolidone. Protein extracts from North Carolina macroalgae were also tested against five bacterial species. Three of these agglutinated bacterial cells. Ulva curvata and Bryopsis plumosa agglutinated all five species. Protein extracts from five species of Florida algae were tested for their effects on mitogenesis in mouse splenocytes and human lymphocytes. Gracilaria tikvahiae HBOI Strain G-5, Ulva rigida and Gracilaria verrucosa HBOI Strain G-16S stimulated mitogenesis in mouse splenocytes, while Gracilaria tikvahiae HBOI Strain G-16stimulated mitogenesis in human lymphocytes.

Ovine bone and marrow derived progenitor cells : isolation, characterization, and osteogenic potential

Recently, research has focused on bone marrow derived multipotent mesenchymal precursor cells (MPC) for their potential clinical use in bone engineering. Prior to clinical application, MPC-based treatment concepts need to be evaluated in preclinical, immunocompetent, large animal models. Sheep in particular are considered a valid model for orthopaedic and trauma related research. However, ovine MPC and their osteogenic potential remain poorly characterized. In the present study, ex vivo expanded MPC isolated from ovine bone marrow proliferated at a higher rate than osteoblasts (OB) derived from tibial compact bone as assessed using standard 2D culture. MPC expressed the respective phenotypic profile typical for different mesenchymal cell populations (CD14-/CD31-/CD45- /CD29+/CD44+/CD166+) and showed a multilineage differentiation potential. When compared to OB, MPC had a higher mineralization potential under standard osteogenic culture conditions and expressed typical markers such as osteocalcin, osteonectin and type I collagen at the mRNA and protein level. After 4 weeks in 3D culture, MPC constructs demonstrated higher cell density and mineralization, whilst cell viability on the scaffolds was assessed >90%. Cells displayed a spindle-like morphology and formed an interconnected network. Implanted subcutaneously into NOD/SCID mice on type I collagen coated polycaprolactone-tricalciumphosphate (mPCL-TCP) scaffolds, MPC presented a higher developmental potential than osteoblasts. In summary, this study provides a detailed in vitro characterisation of ovine MPC from a bone engineering perspective and suggests that MPC provide promising means for future bone disease related treatment applications.

Maternal administration of erythromycin fails to eradicate intrauterine ureaplasma infection in an ovine model

Erythromycin is the standard antibiotic used for treatment of Ureaplasma species during 3 pregnancy; however, maternally administered erythromycin may be ineffective at eliminating 4 intra-amniotic ureaplasma infections. We asked if erythromycin would eradicate intra-amniotic 5 ureaplasma infections in pregnant sheep. At 50 days of gestation (d, term=150d) pregnant ewes 6 received intra-amniotic injections of erythromycin-sensitive U. parvum serovar 3 (n=16) or 10B 7 medium (n=16). At 100d, amniocentesis was performed; five fetal losses (ureaplasma group: 8 n=4; 10B group: n=1) had occurred by this time. Remaining ewes were allocated into treatment 9 subgroups: medium only (M, n=7); medium and erythromycin (M/E, n=8); ureaplasma only (Up, 10 n=6) or ureaplasma and erythromycin (Up/E, n=6). Erythromycin was administered intra11 muscularly (500 mg), eight-hourly for four days (100d-104d). Amniotic fluid samples were 12 collected at 105d. At 125d preterm fetuses were surgically delivered and specimens were 13 collected for culture and histology. Erythromycin was quantified in amniotic fluid by liquid 14 chromatography-mass spectrometry. Ureaplasmas were isolated from the amniotic fluid, 15 chorioamnion and fetal lung of animals from the Up and Up/E groups, however, the numbers of 16 U. parvum recovered were not different between these groups. Inflammation in the 17 chorioamnion, cord and fetal lung was increased in ureaplasma-exposed animals compared to 18 controls, but was not different between the Up and Up/E groups. Erythromycin was detected in 19 amniotic fluid samples, although concentrations were low (<10-76 ng/mL). This study 20 demonstrates that maternally administered erythromycin does not eradicate chronic, intra- amniotic ureaplasma infections or improve fetal outcomes in an ovine model, potentially due to 22 the poor placental passage of erythromycin.

Spatial and temporal variations of mechanical properties and mineral content of the external callus during bone healing

After bone fracture, various cellular activities lead to the formation of different tissue types, which form the basis for the process of secondary bone healing. Although these tissues have been quantified by histology, their material properties are not well understood. Thus, the aim of this study is to correlate the spatial and temporal variations in the mineral content and the nanoindentation modulus of the callus formed via intramembranous ossification over the course of bone healing. Midshaft tibial samples from a sheep osteotomy model at time points of 2, 3, 6 and 9 weeks were employed. PMMA embedded blocks were used for quantitative back scattered electron imaging and nanoindentation of the newly formed periosteal callus near the cortex. The resulting indentation modulus maps show the heterogeneity in the modulus in the selected regions of the callus. The indentation modulus of the embedded callus is about 6 GPa at the early stage. At later stages of mineralization, the average indentation modulus reaches 14 GPa. There is a slight decrease in average indentation modulus in regions distant to the cortex, probably due to remodelling of the peripheral callus. The spatial and temporal distribution of mineral content in the callus tissue also illustrates the ongoing remodelling process observed from histological analysis. Most interestingly the average indentation modulus, even at 9 weeks, remains as low as 13 GPa, which is roughly 60% of that for cortical sheep bone. The decreased indentation modulus in the callus compared to cortex is due to the lower average mineral content and may be perhaps also due to the properties of the organic matrix which might be different from normal bone.

Mechanical Behavior of Articular Cartilage After Osteochondral Autograft Transfer in an Ovine Model

BACKGROUND: Grafting of autologous hyaline cartilage and bone for articular cartilage repair is a well-accepted technique. Although encouraging midterm clinical results have been reported, no information on the mechanical competence of the transplanted joint surface is available. HYPOTHESIS: The mechanical competence of osteochondral autografts is maintained after transplantation. STUDY DESIGN: Controlled laboratory study. METHODS: Osteochondral defects were filled with autografts (7.45 mm in diameter) in one femoral condyle in 12 mature sheep. The ipsilateral femoral condyle served as the donor site, and the resulting defect (8.3 mm in diameter) was left empty. The repair response was examined after 3 and 6 months with mechanical and histologic assessment and histomorphometric techniques. RESULTS: Good surface congruity and plug placement was achieved. The Young modulus of the grafted cartilage significantly dropped to 57.5% of healthy tissue after 3 months (P < .05) but then recovered to 82.2% after 6 months. The aggregate and dynamic moduli behaved similarly. The graft edges showed fibrillation and, in some cases (4 of 6), hypercellularity and chondrocyte clustering. Subchondral bone sclerosis was observed in 8 of 12 cases, and the amount of mineralized bone in the graft area increased from 40% to 61%. CONCLUSIONS: The mechanical quality of transplanted cartilage varies considerably over a short period of time, potentially reflecting both degenerative and regenerative processes, while histologically signs of both cartilage and bone degeneration occur. CLINICAL RELEVANCE: Both the mechanically degenerative and restorative processes illustrate the complex progression of regeneration after osteochondral transplantation. The histologic evidence raises doubts as to the long-term durability of the osteochondral repair.

Osteoclastic activity begins early and increases over the course of bone healing

Osteoclasts are specialised bone-resorbing cells. This particular ability makes osteoclasts irreplaceable for the continual physiological process of bone remodelling as well as for the repair process during bone healing. Whereas the effects of systemic diseases on osteoclasts have been described by many authors, the spatial and temporal distribution of osteoclasts during bone healing seems to be unclear so far. In the present study, healing of a tibial osteotomy under standardised external fixation was examined after 2, 3, 6 and 9 weeks (n = 8) in sheep. The osteoclastic number was counted, the area of mineralised bone tissue was measured histomorphometrically and density of osteoclasts per square millimetre mineralised tissue was calculated. The osteoclastic density in the endosteal region increased, whereas the density in the periosteal region remained relatively constant. The density of osteoclasts within the cortical bone increased slightly over the first 6 weeks, however, there was a more rapid increase between the sixth and ninth weeks. The findings of this study imply that remodelling and resorption take place already in the very early phase of bone healing. The most frequent remodelling process can be found in the periosteal callus, emphasising its role as the main stabiliser. The endosteal space undergoes resorption in order to recanalise the medullary cavity, a process also started in the very early phase of healing at a low level and increasing significantly during healing. The cortical bone adapts in its outward appearance to the surrounding callus structure. This paradoxic loosening is caused by the continually increasing number and density of osteoclasts in the cortical bone ends. This study clearly emphasises the osteoclastic role especially during early bone healing. These cells do not simply resorb bone but participate in a fine adjusted system with the bone-producing osteoblasts in order to maintain and improve the structural strength of bone tissue.

CYR61 (CCN1) protein expression during fracture healing in an ovine tibial model and its relation to the mechanical fixation stability

The formation of new blood vessels is a prerequisite for bone healing. CYR61 (CCN1), an extracellular matrix-associated signaling protein, is a potent stimulator of angiogenesis and mesenchymal stem cell expansion and differentiation. A recent study showed that CYR61 is expressed during fracture healing and suggested that CYR61 plays a significant role in cartilage and bone formation. The hypothesis of the present study was that decreased fixation stability, which leads to a delay in healing, would lead to reduced CYR61 protein expression in fracture callus. The aim of the study was to quantitatively analyze CYR61 protein expression, vascularization, and tissue differentiation in the osteotomy gap and relate to the mechanical fixation stability during the course of healing. A mid-shaft osteotomy of the tibia was performed in two groups of sheep and stabilized with either a rigid or semirigid external fixator, each allowing different amounts of interfragmentary movement. The sheep were sacrificed at 2, 3, 6, and 9 weeks postoperatively. The tibiae were tested biomechanically and histological sections from the callus were analyzed immunohistochemically with regard to CYR61 protein expression and vascularization. Expression of CYR61 protein was upregulated at the early phase of fracture healing (2 weeks), decreasing over the healing time. Decreased fixation stability was associated with a reduced upregulation of the CYR61 protein expression and a reduced vascularization at 2 weeks leading to a slower healing. The maximum cartilage callus fraction in both groups was reached at 3 weeks. However, the semirigid fixator group showed a significantly lower CYR61 immunoreactivity in cartilage than the rigid fixator group at this time point. The fraction of cartilage in the semirigid fixator group was not replaced by bone as quickly as in the rigid fixator group leading to an inferior histological and mechanical callus quality at 6 weeks and therefore to a slower healing. The results supply further evidence that CYR61 may serve as an important regulator of bone healing.

The course of bone healing is influenced by the initial shear fixation stability

Fracture healing is influenced by fixation stability and experimental evidence suggests that the initial mechanical conditions may determine the healing outcome. We hypothesised that mechanical conditions influence not only the healing outcome, but also the early phase of fracture healing. Additionally, it was hypothesised that decreased fixation stability characterised by an increased shear interfragmentary movement results in a delay in healing. Sixty-four sheep underwent a mid-shaft tibial osteotomy which was treated with either a rigid or a semi-rigid external fixator. Animals were sacrificed at 2, 3, 6 and 9 weeks postoperatively and the fracture callus was analysed using radiological, biomechanical and histological techniques. The tibia treated with semi-rigid fixation showed inferior callus stiffness and quality after 6 weeks. At 9 weeks, the calluses were no longer distinguishable in their mechanical competence. The calluses at 9 weeks produced under rigid fixation were smaller and consisted of a reduced fibrous tissue component. These results demonstrate that the callus formation over the course of healing differed both morphologically and in the rate of development. In this study, we provide evidence that the course of healing is influenced by the initial fixation stability. The semi-rigid fixator did not result in delayed healing, but a less optimal healing path was taken. An upper limit of stability required for successful healing remains unknown, however a limit by which healing is less optimal has been determined.