The thermostable β-1,3-1,4-glucanase from Clostridium thermocellum improves the nutritive value of highly viscous barley-based diets for broilers

1. Microbial β-1,3-1,4-glucanases improve the nutritive value of barley-based diets for poultry by effectively decreasing the degree of polymerisation of the anti-nutritive soluble β-glucans. Glycoside hydrolases (GHs) acting on recalcitrant polysaccharides display a modular architecture comprising a catalytic domain linked to one or more non-catalytic Carbohydrate-Binding Modules (CBMs).

2. GHs and CBMs have been classified in different families based on primary structure similarity (see CAZy webpage at http://www.cazy.org). The role of CBMs is to anchor the appended GHs into their target substrates, therefore eliciting the efficient hydrolysis of structural carbohydrates.

3. Here we describe the biochemical properties of the family 16 GH from Clostridium thermocellum, termed CtGlc16A. CtGlc16A is a thermostable enzyme that specifically acts on β-1,3-1,4-glucans with a remarkable catalytic activity (38000?U/mg protein).

4. CtGlc16A, individually or fused to the family 11 β-glucan-binding domain of cellulase CtLic26A-Cel5E of C. thermocellum, was used to supplement a highly viscous barley-based diet for broilers.

5. The data showed that birds fed on diets supplemented with the recombinant enzymes displayed an improved performance when compared with birds given diets not supplemented with exogenous enzymes. However, inclusion of the non-catalytic CBMs had no influence on the capacity of CtGlc16A to reduce the anti-nutritive effects of soluble β-1,3-1,4-glucans.

6. The data suggest that at elevated dosage rates, CBMs might be unable to potentiate the catalytic activity of appended catalytic domains; this effect may only be revealed when feed enzymes are incorporated at low levels.     

Chemical composition and nutritional value in vitro of mechanically‐deboned poultry meat

1. The composition by proximate analysis, amino acid content and digestibility by mammalian proteolytic enzymes of pastes from mechanically‐deboned chicken breasts wings, necks and backs, and hand‐deboned breast were measured. Values obtained were used to calculate indices of nutritional value.

2. All pastes compared favourably with similar protein materials used for human food. Mechanical deboning did not alter the nutritional indices.

3. The protein quality and digestibility of the breast were somewhat higher than those for other parts.     

Increased small intestinal fermentation is partly responsible for the anti‐nutritive activity of non‐starch polysaccharides in chickens

1. The mechanism of the anti‐nutritive activities of soluble non‐starch polysaccharides (NSPs) in broiler diets was investigated with emphasis on the inter‐relationship between viscosity and fermentation along the gut. Isolated soluble NSP were added to a control diet to effect high gut viscosity, and in vivo depolymerisation of the NSP was achieved using a commercial glycanase.

2. Addition of soluble NSPs significantly (P<0.01) increased gut viscosity, reduced the AME of the diet and depressed the growth and FCE of the birds. Enzyme supplementation of the NSP‐enriched diet reversed the adverse effects, increasing (P< 0.01) weight gain, FCE and AME. Comparisons of the viscosities (mPa) in birds fed on the NSP‐enriched diet and the same diet supplemented with enzyme were respectively: 11–9 v. 2–3 in the duodenum; 78–3 v. 4–4 in the jejunum and 409–3 v. 10–8 in the ileum.

3. Caecal volatile fatty acid concentration was markedly (P<0.01) elevated by enzyme supplementation, whereas ileal fermentation was inhibited.

4. Microscopic examination revealed that, among birds fed on the NSP‐enriched diet, there had been extensive small intestinal fermentation, which was eliminated by the enzyme supplementation.

5. Addition of a synthetic antibiotic (Amoxil) had no beneficial effects.

6. The current study demonstrated that increased fermentation occurs in the small intestine when a large amount of viscous NSPs is present in the diet and this is detrimental to the performance and well‐being of poultry.     

Variation among individual white‐leghorn hens in the concentration of minerals in the albumen and yolk content of their eggs

1. The mineral composition of the albumen and yolk was determined in several eggs from each of a number of individual hens from the same White‐Leghorn strain. X‐ray fluorescent spectrometry was used to undertake two independent series of analyses. A total of 8 minerals (calcium, chlorine, iron, magnesium, phosphorus, potassium, sodium, sulphur) were included in the analyses of the yolk and the same minerals, but excluding iron (which is present in only small amounts), in the albumen.

2. There was considerable variation between individuals in the mineral concentration in their eggs (coefficients of variation ranged between 3.8% for sodium to 19.9% for calcium in the albumen, and between 4.3% for phosphorus to 11.8% for iron in the yolk).

3. At the same time, the moderately high repeatability of mineral concentration (t = 0.4?0.6) in successive eggs from the the same hen for several of the minerals analysed is indicative of some positive control by the hen of the mineral composition of her eggs.

4. There was a highly significant correlation (P≤ 0.001) between the mean concentration of potassium in the albumen and the hatchability of the eggs, supporting the claim that a deficiency of potassium in the egg could be the basis of some failures in hatchability.

5. The study also revealed variation among individual birds in the concentration of iron in the yolk which was negatively correlated (P≤0.01) with hatchability. No clear basis could be suggested for this variation among individual birds.     

Increased levels of the 14-3-3 η and γ proteins in the synovial fluid of dogs with unilateral cranial cruciate ligament rupture

The present study investigated whether the 14-3-3 η and γ proteins, which are potent matrix metalloprotease (MMP) stimulators, are detectable in the synovial fluid of dogs with cranial cruciate ligament rupture (CCLR). Synovial fluid samples from 7 dogs with unilateral CCLR and control samples from 4 dogs without a history of any joint inflammation or any other abnormalities underwent Western blot analysis for the 14-3-3 η, γ, and σ proteins as well as MMP-1 and MMP-3. Craniocaudal and lateral radiographic projections of the stifle joint were evaluated for the presence and severity of 13 specific radiographic markers of osteoarthritis and graded numerically. The Spearman method was used to detect any correlation between the 14-3-3-η level in the synovial fluid and the radiograph-based grade. The η isoform was present only in the samples from the dogs with CCLR. The levels of 14-3-3-γ, MMP-1, and MMP-3 were significantly higher in the samples from the dogs with CCLR than in the control samples (P < 0.05). However, there was no significant difference between the CCLR and control samples in the level of the σ isoform. The Spearman method showed a significant correlation between the 14-3-3-η level in the synovial fluid and the presence of either patellar osteophytes or lateral or medial (or both) condylar periarticular osteophytes (P < 0.05). The MMP stimulatory effect of the 14-3-3 η and γ isoforms may be the reason for the high levels of MMP-1 and MMP-3 observed. Thus, 14-3-3 proteins, especially the η isoform, may be important markers of osteoarthritis caused by CCLR.     

Accuracy of capillary blood 3-β-hydroxybutyrate determination for the detection and treatment of canine diabetic ketoacidosis

In human medicine, diagnosis of diabetic ketoacidosis (DKA) is usually based on measurement of capillary 3-β-hydroxybutyrate (3-HB) with a hand held ketone sensor. This study was conducted to determine if measurement of capillary 3-HB could be useful for the diagnosis and monitoring of canine DKA. Fifteen dogs with diabetic ketosis and 10 with DKA were evaluated. Paired measurements of 3-HB of capillary and venous blood samples were analysed by the electrochemical sensor and reference method. Use of capillary 3-HB measurement during DKA management was then evaluated through simultaneous measurements of capillary 3-HB, urinary AcAc and venous blood gas analysis. Good agreement between capillary and venous 3-HB measurement was detected by the electrochemical sensor and reference method. Monitoring treatment of DKA revealed a significant correlation between capillary 3-HB and acidosis markers, while no significant correlation was observed between AcAc and acidosis markers. A cut-off value of capillary blood 3-HB >3.8 mmol/L for diagnosis of DKA resulted in 70% and 92% sensitivity and specificity. The electrochemical sensor accurately measures 3-HB concentration in both capillary and venous blood samples, is accurate in diagnosing canine DKA, and appears to reflect the patient''s metabolic status during DKA treatment.     

The influence of in ovo injection with the prebiotic DiNovo~? on the development of histomorphological parameters of the duodenum,body mass and productivity in large-scale poultry production conditions

Background: Among various feed additives currently used in poultry nutrition, an important role is played by bioactive substances, including prebiotics. The beneficial effect of these bioactive substances on the gastrointestinal tract and immune system give rise to improvements in broiler health and performance nutrition, thus increasing the productivity of these birds. An innovative method for introducing bioactive substances into chickens is the in ovo injection into eggs intended for hatching. The aim of the study was to evaluate the development of histomorphological parameters of the duodenum and productivity in chickens injected in ovo with the prebiotic DiNovo~?(extract of Laminaria species of seaweed, BioA tlantis Ltd., Ireland) on d 12 of incubation, under large-scale, high density poultry production conditions.Results: There was no significant impact of the injection of DiNovo~? prebiotic on the production parameters of broiler chickens(body weight, FCR, EBI and mortality) obtained on d 42 of rearing. No significant impact of the DiNovo~? injection on the duodenum weight and length was observed, as wel as on the CSA, diameter and muscular layer thickness of the duodenum. The in ovo injection of DiNovo~? significantly increased the width of the duodenal vil i(P 0.05) and crypt depth(P 0.01) of chickens on d 21 of rearing. Other histomorphological parameters of duodenal vil i at d 42 of chickens rearing such as: the height, width, and cross section area of vil i were significantly greater in chickens from the control group compared to those from the DiNovo~? group(P 0.05 and P 0.01).Conclusions: In conclusion, this study demonstrates that injection of DiNovo~? prebiotic into the air chamber of egg significantly influences the histomorphological parameters on d 21 of rearing without negatively affecting productivity in chickens at the end of rearing.     

Postpartum toll-like receptors and β-defensin 5 mRNA levels in the endometrium of Holstein cows

Toll-like receptors (TLRs) and β-defensins are important components of the innate immune system. This study aimed to evaluate endometrial mRNA levels of TLRs (1/6, 2, 4, and 5) and β-defensin 5 in Holstein cows by quantitative real time RT-PCR. Uterine biopsies were performed from 6 to 12 h after parturition, and cows were divided into two groups: (i) cows with placental retention and clinical signs of uterine infection until 45 days postpartum (n=10) or (ii) cows with normal puerperium (n=10). All cows had detectable levels of TLRs and β-defensin 5 mRNAs, but these levels did not differ between groups (P>0.05). Levels of TLR4 mRNA had a positive and significant correlation with the time required for uterine involution in both groups.     

Evaluation of methanol‐grown bacteria and hydrocarbon‐grown yeast as sources of protein for poultry: Performance of broilers during the finishing period

1. Three trials were carried out with male broilers during the 5‐ to 8‐week period, for the evaluation of two types of single‐cell protein (SCP) in practical‐type broiler finisher diets. The SCP tested were: Pruteen, produced from methanol‐utilising bacteria, and a Lavera‐type yeast which utilises the normal paraffins of heavy gas oil.

2. The inclusion of 90 to 100 g of either type of SCP/kg of the diet caused a 2 to 3% reduction in weight gain, a 3 to 5% decrease in food consumption, and up to a 2% improvement in food utilisation. The reduced growth rate could be fully explained by the decrease in food intake, with essentially no effect on food : gain ratio.

3 In diets containing Pruteen, arginine was second limiting after the sulphur amino acids.

4 An odd‐numbered fatty acid (17 carbons, unsaturation unknown) of Pruteen was not found in the carcass lipids of broilers fed on a diet containing 120 g Pruteen/kg.     

The Analysis for the Single Nucleotide Polymorphism of TGF-β1 and Its Association with Reproduction in Different Sheep Breeds

A single nucleotide polymorphism(SNP)of 805 bp region in the intron 6 of transforming growth factor β1(TGF-β1)gene was identified by polymerase chain reaction-single-strand composition polymorphism(PCR-SSCP)in 196 sheep among Small-tailed Han sheep,Tong sheep,Tan sheep and Oula sheep.Comparative sequence analysis of cloned products revealed an AGAC deletion at 294 bases upstream of exon 7 of the TGF-β1 gene(site 14201 in gi76871756).Statistical results of the genotype and allele frequencies in different breeds showed that genotype AB was dominant in the Small-tailed Han sheep.Genotype BB,however,was in majority in low-fecundity sheep.The results of a Chi-square test indicated that all the populations were in Hardy-Weinberg equilibrium.     

Influence of in ovo injection of disaccharides, glutamine and β-hydroxy-β-methylbutyrate on the development of small intestine in duck embryos and neonates

1. The objective of the present study was to examine the effect of in ovo injection of disaccharides (DS), disaccharides and glutamine (DS + Gln) or disaccharides and β-hydroxy-β-methylbutyrate (DS + HMB) at d 23 of incubation on the development of the small intestine. 2. In DS + Gln-injected ducks, the greatest relative small intestine mass and muscularis layer thickness among 4 treatments was observed from d 25 of incubation to 7 d of age. 3. Jejunal sucrase activity in DS-injected ducks was significantly greater than in controls at hatch and on d 7. 4. In DS + HMB-treated ducks, a tendency toward slightly higher jejunal DNA concentration was observed throughout the experiment. 5. Greater body weight was found in DS + Gln and DS + HMB treated ducks in the first two weeks. However, there was no significant difference in the market weight (35 d) of ducks among the 4 treatments. 6. The results of present study suggest that administering disaccharides and Gln, or disaccharides and HMB, to the duck embryos exerted a beneficial effect on the early development of small intestine and on growth performance.     

Intra- and interlaboratory variability of allergen-specific IgE levels in atopic dogs in three different laboratories using the Fc-ɛ receptor testing

The IgE specific Fc-? receptor test is widely used for serological evaluation of allergic canine patients. Aim of the study was to evaluate intra- and interlaboratory variability in three independent laboratories. Duplicate serum aliquots from 15 atopic dogs were submitted simultaneously to 3 laboratories (LA, LB, LC), which subsequently performed the test in a blinded fashion. LA and LC analysed sera for 35 allergens and expressed the results as optical density units (OD), while LB analysed 15 allergens and provided reaction grades (RG). Results were compared with three factorial ANOVA. Intralaboratory variability was evaluated by calculating the dispersion factor for data of LA and LC and standard deviation (SD) for LB. Correlation coefficients (r) were used to evaluate interlaboratory variability. Depending on the allergens, intralaboratory dispersion factor for LA and LC varied from 1.15 to 6.63 and 1.19 to 6.17, respectively. In LB the SD varied between 0 and 0.6 RG. Regarding interlaboratory variability, correlation of OD values between LA and LC was excellent (r > 0.8, p < 0.001) for 13 of 35 allergens. There was a significant difference in OD results for 9 allergens. The results were significantly correlated (r > 0.8, p < 0.001) for all except 3 allergens (Dermatophagoides pteronyssinus, mixed grasses, nettle). As far as negative/positive results were concerned, intralaboratory differences were 3.14% and interlaboratory differences were 4.76%.     

The preliminary evaluation of grasses and legumes for use in the high‐rainfall sandveld of Rhodesia

Uittreksel

In hierdie studie is ondersoek ingestel na die invloed van vier belankrike bosveldbome, naamlik, Acacia Senegal, Acacia tortilis, Boscia albitrunca en Combretum apiculatum op die verspreiding en en produktiwiteit van Panicum maximum.

Die studie is in twee fases uitgevoer: deur middel van veldproewe in die Laar‐ Krokodilrivier‐vallei, noord‐wes van Thabazimbi, en in die glasbuise van die Potchefstroomse Universiteit.

In die veld is opnames deur middel van strookpersele uitgevoer om die verspreiding van Panicum maximum in assosiasie met bogenoemde boomsoorte na te gaan.

In die glasbuise is die groeivermoë van Panicum maximum in grondmonsters wat onder die vier boomsoorte versamel is met dié van grondmonsters wat in oop dele versamel is vergelyk. Die invloed van ligte graad van beskaduïng en twee verskillende vogpeile op die groeivermoë van die proef plante is ter‐selfdertyd ondersoek.

‘n Proef is ook uitgevoer om die groeivermoë van Panicum maximum by toedienings van verskillende peile N, P en K by grond wat in oop dele versamel is te ondersoek.     

Evidence for vascular and enzymatic events in the pathophysiology of acute laminitis: which pathway is responsible for initiation of this process in horses?

To date, there is a substantial amount of data to support the hypotheses that vascular and enzymatic changes are ongoing in experimental laminitis. Furthermore, there is substantial in vitro evidence that the enzymatic changes weaken the dermo-epidermal attachments leading to mechanical failure of the hoof-bone interface of the equine digit. However, investigators of both the vascular and enzymatic theories have, to date, been unable to substantiate the effects of these pathophysiological changes in vivo on laminar tissues of horses afflicted with experimentally induced or naturally acquired laminitis. In addition, the effects of laminitis-inducing treatment have not been prevented or reversed by treatment with an MMP inhibitor or a vasoactive antagonist. It is possible that there is simultaneous activation of the vascular and enzymatic pathways and/or other inflammatory processes. Moreover, the third theory involving mechanical factors cannot be discounted simply because strong evidence for vascular and enzymatic changes exists. It is common for horses with severe musculoskeletal disease affecting weightbearing on a limb to develop laminitis in the contralateral limb. It remains to be determined what factors are responsible for initiation of laminitis in these individuals. Evidence has not been presented that precludes the possibility of coincident occurrence of vascular and enzymatic changes. In fact, many of the inflammatory mediators (e.g. interleukin-1beta) found in laminitic tissues can concurrently stimulate synthesis of vasoactive substances and activate MMPs. Because enzymatic action on proteins is largely dependent on the concentrations of proteins and enzyme, the enzymatic theory is not dependent upon increased delivery of enzymes via increased capillary flow. Likewise, because vascular changes can alter tissue function via increased capillary flow and oedema formation, the vascular theory is not dependent upon decreased capillary flow. It is true that naturally acquired laminitis is widely variable in severity and predisposing diseases. Therefore, most probably there are multiple mechanisms involved in the initiation and propagation of the pathophysiologic cascade(s) and, therefore, successful intervention will necessitate multiple treatment modalities.     

Assessment of Molasses–Urea Blocks for Goat and Sheep Production in the Sultanate of Oman: Intake and Growth Studies

The goal of this study was to develop a molasses–urea block (MUB) for purposes of supplementing trace minerals to domestic ruminant livestock in Oman. To accomplish this, the utility of molasses and date syrup as fermentable energy sources, of straw, date flakes and wheat bran as fibre sources, and of cement and lime as binders were evaluated. The proportion of cement needed for adequate hardening of the block was also studied. Molasses- and date syrup-based blocks hardened equally well. However, the higher cost of date syrup precluded its use. Wheat straw yielded a low-density block that hardened slowly. Date fibre retained moisture and hardened extremely slowly. Wheat bran-based blocks hardened quickly and yielded dense blocks. Hence, wheat bran was judged to be the superior source of fibre. Lime did not effectively bind the blocks. A cement content of 15% allowed hardening of the blocks within 2–3 weeks. A level of 10% cement in the block reduced the hardening rate by about 50%. Sheep and goats consumed both the straw- and wheat bran-based blocks but at different rates. Consumption of the straw-based block by sheep ranged from 50 to 179 g/head per day, whereas the denser wheat bran-based block was consumed at a rate of 8–20 g/head per day. Consumption of the straw-based block by goats was low (8 g/head per day) compared to that of wheat bran-based blocks (16–24 g/head per day). On the basis of the intake of the bran-based block by sheep, a block was designed that would provide approximately 50% of an animal's trace mineral requirements per day. This block consisted of 45% molasses, 10% urea, 5% trace minerals, 2.5% NaCl, 22.5% wheat bran and 15% cement. Sheep consuming this block gained more weight than sheep fed a conventional mineral block or sheep receiving no mineral supplementation. MUBs are inexpensive (9.5 US cents/kg). We conclude that MUBs have utility for providing trace elements in ruminant diets.     

Effectiveness of vaccines and vaccination programs for the control of foot-and-mouth disease in Uganda, 2001–2010

Foot-and-mouth disease (FMD) is a highly contagious disease of cloven-hoofed animals. In Uganda, FMD outbreaks are mainly controlled by ring vaccination and restriction of animal movements. Vaccination stimulates immunity and prevents animals from developing clinical signs which include lameness, inappetence, and decreased production. Ring vaccination and restriction of animal movements have, however, not successfully controlled FMD in Uganda and outbreaks reoccur annually. The objective of this study was to review the use of FMD virus (FMDV) vaccines and assess the effectiveness of vaccination programs for controlling FMD in Uganda (2001–2010), using retrospective data. FMD vaccine distribution patterns in Uganda (2001–2010) matched occurrence of outbreaks with districts reporting the highest number of outbreaks also receiving the largest quantity of vaccines. This was possibly due to “fire brigade” response of vaccinating animals after outbreaks have been reported. On average, only 10.3 % of cattle within districts that reported outbreaks during the study period were vaccinated. The average minimum time between onset of outbreaks and vaccination was 7.5 weeks, while the annual cost of FMDV vaccines used ranged from US $58,000 to 1,088,820. Between 2001 and 2010, serotyping of FMD virus was done in only 9/121 FMD outbreaks, and there is no evidence that vaccine matching or vaccine potency tests have been done in Uganda. The probability of FMDV vaccine and outbreak mismatch, the delayed response to outbreaks through vaccination, and the high costs associated with importation of FMDV vaccines could be reduced if virus serotyping and subtyping as well as vaccine matching were regularly done, and the results were considered for vaccine manufacture.     

Evaluation of untreated and urea molasses–treated finger millet (Eleusine coracana) straw and lowland bamboo (Oxytenanthera abyssinica) leaf hay on nutritive values and the performance of Gumuz sheep in Ethiopia

Tropical Animal Health and Production - The experiment was conducted to evaluate effects of untreated and urea molasses–treated finger millet straw and lowland bamboo leaf hay feeding on...