The evolution and application of enzymes in the animal feed industry: the role of data interpretation

ABSTRACT

1. Enzymes have been used commercially for nearly 40 years and save significant costs through sparing of expensive nutrients but the mechanism by which this is achieved is still debated.

2. The research focused on non-starch polysaccharidase (NSPase) enzymes is used as an example of where greater progress could have been made if the details of the work had been described more fully and the analysis of the data generated had been broader in scope and more critical.

3. Lack of standardisation of the details presented in the materials and methods has been identified as a significant barrier to meaningful retrospective analysis and thus limits advances in the understanding of the mode of action of these enzymes.

4. The identity of the enzyme employed and its activity is often lacking, and more importantly the purity is rarely disclosed. Contaminant activities which are neither listed nor assayed could play a significant role in the responses observed.

5. The dose optimum of most enzymes is often considerably higher than that employed in most studies. Thus studies claiming synergy between two ‘activities’ should ensure that the response is not related to each enzyme simply augmenting the dose of just one activity in the finished feed. This is a common problem, and coupled with the lack of factorial experiments to justify the presence of each enzyme in a multi-enzyme product, it is not surprising that there is still debate as to whether single or multi-enzymes are best suited poultry rations.

6. The three proposed mechanisms for NSPases (viscosity, cell wall and prebiotic) are discussed, and along with their strengths and weaknesses it is suggested that a re-evaluation of each is needed. Viscosity may have to be re-evaluated as being a function not only of the cereal being fed, but of the age of the animal as well. The cell wall theory as described is poorly modelled in vitro and hence the validity of these data is questioned. The prebiotic theory may need significant modification as it appears that the quantities of oligomers produced are insufficient to generate the additional volatile fatty acids (VFA)’s reported. It is likely that all three mechanisms play a role in the responses observed, but the prebiotic mechanism probably plays by far the most important part in low viscosity diets.

7. Future research would be improved if it considered all potential mechanisms when designing a trial. Significant failings are apparent as a result of adherence to tenets in explanation of the results. Most importantly, it should be emphasised that a hypothesis is there to be tested, not defended.     

Effect of dietary inclusion of date seed (<Emphasis Type="Italic">Phoenix dactylifera</Emphasis> L.) on intake,digestibility, milk production,and milk fatty acid profile of Holstein dairy cows

The objective of this experiment was to investigate the influence of ground date seed (GDS) on intake, digestibility, and milk yield and milk fatty acid (FA) composition of lactating Holstein cows. The experimental design was a 4?×?4 replicated Latin square with eight lactating dairy cows with an average milk production of 35.5?±?1.5 kg and 75?±?5 days in milk (DIM). Dairy cows were fed one of the four treatments contained 0, 2, 4, and 6% of diet dry matter (DM) GDS in replacement of wheat bran. All diets contained the same amount of forages (alfalfa hay and corn silage). Dietary treatments had no effect on DM intake (DMI), total tract apparent digestibility, milk yield, and milk composition. Increasing GDS linearly decreased concentration of C13:0 and increased cis-9 C14:1 and trans-11 C18:1 (vaccenic acid) (P?<?0.05). A linear tendency for more C16:1 content in milk fat was observed with increasing GDS (P?=?0.06). Feeding GDS resulted in a linear decrease (P?<?0.01) in saturated FA (SFA) but increased milk fat monounsaturated FA (MUFA) and trans FA (TFA) (P?<?0.05). Therefore, low levels of GDS (up to 6%) in the diet of Holstein dairy cows can beneficially modify milk FA composition without any adverse effects on intake, digestibility, and milk yield.     

Genetic evaluation of growth in farmers’ flocks of Madras Red sheep under long-term selection in a group breeding scheme

The Network Project on Sheep Improvement (NWPSI)–Madras Red field unit is a group breeding scheme involving 198 farmers’ flocks of Madras Red sheep in which selection for growth traits and rotation of rams have been practised for over two decades. Growth data collected from these flocks were used to evaluate the performance and understand the direct and expected responses to selection based on genetic parameters. The body weight at birth (BW), weaning weight (WW), 6-month weight (6W), 9-month weight (9W), 12-month weight (YW), pre-weaning average daily gain (ADG1, birth to 3 months), post-weaning ADG2 (3–6 months), ADG3 (6–9 months), ADG4 (9–12 months) and ADG5 (3–12 months) were 2.67, 10.05, 14.56, 18.36, 21.36, 80.13, 49.05, 43.00, 34.21 and 41.18 g, respectively. Univariate analyses were carried out using animal and sire models to estimate variance components. Heritability obtained from animal model for BW was 0.36 and the values for other body weight traits were almost unity. Heritability estimate for pre-weaning ADG1 was 0.31. Very high genetic variability was observed in spite of long-term selection and this sustenance of variability is one of the main advantages of a group breeding scheme, combining several flocks of smaller size. An increasing genetic and phenotypic trend was noticed for almost all the traits studied. The expected responses calculated based on genetic parameters also indicated scope for improvement.     

Male-to-female aggression in cage-housed common pheasants (Phasianus colchicus) during the breeding season was not related to male plasma testosterone level

1. The aim of this study was to investigate if male-to-female aggression of common pheasants in the course of the breeding season was related to the concentration of plasma testosterone and/or other biochemical plasma indicators in male pheasants housed in breeding cages. The influence of season on the concentration of testosterone and biochemical indicators was also investigated.

2. Males were divided into non-aggressive and aggressive groups during the breeding season based on ethological evaluation. At the beginning, in the middle and at the end of the breeding season, a blood sample was taken from all males on the same day and the concentration of selected biochemical indicators and the total circulating testosterone in the plasma were determined.

3. Male-to-female aggression during the breeding season of pheasants was not influenced by the total plasma testosterone of males.

4. The concentration of total plasma testosterone in males decreased gradually during the breeding season.

5. Male-to-female aggression of pheasants did not have a significant effect on any of the assessed biochemical indicators.

6. The influence of the breeding season affected the activities of alanine aminotransferase and aspartate aminotransferase as well as the concentrations of glucose, magnesium, potassium and chloride in the blood plasma of cage-housed male pheasants.     

The Impact of Natural Deep Eutectic Solvents and Extraction Method on the Co-Extraction of Trace Metals from Fucus vesiculosus

In recent years, natural deep eutectic solvents (NADES) have been widely investigated for the extraction of food and medicinal plants as well as seaweeds. However, the ability of NADES for trace elements co-extraction from natural sources is not well investigated. The aim of this study was to investigate the ability of common NADES for trace elements co-extraction from Fucus vesiculosus. All of the tested NADES did not recover As and Co (concentration <LOQ). Moreover, all of the tested NADES provided a low recovery (<9%) of Ba, Ca, Fe, Mg, Mn, Sr, and Zn. The method of extraction had not shown a statistically significant effect on the co-extraction of all elements (excluding Ba and Ca). In contrast, the water content in NADES was significantly affected on the recovery of Ba, Ca, Mg, Mn, Sr, and Zn. The recovery of Al and Cr was relatively high and considerably varied (from 1.5 to 59.9%). NADES comprising lactic acid:glucose:H₂O (5:3:1) provided the lowest contents of all elements, and the highest extracted amounts were obtained employing water contents of 60–80%. The calculated daily intake of all the elements contained in NADES extracts were less than the daily dose risk estimators. The hazard quotients, hazard indexes, and carcinogenic risk calculated for all trace elements and their combination were considerably less than 1. This evidences no health risk, and carcinogenic risk after topical application of all studied NADES. For the first time, the results of the current study demonstrated that NADES extracts of F. vesiculosus contain a lower amount of trace metals and are safer than the extracts obtained with water and 70% acetone. This indicates a significant advantage for NADES compared with the other solvents.     

Utilization of Sake lees as Broiler Feedstuff and its Effects on Growth Performance and Intestinal Immunity

Increasing food loss and waste (FLW) is a global problem, and efforts are being made to use waste food as potential livestock feed material. The amount of self-supplied feed is lower in Japan than in other countries, and the government recommends FLW use for animal feed. Sake (Japanese rice wine) is a traditional alcoholic beverage. During the sake manufacturing process, large amounts of squeezed solids or “lees” (sake lees) are generated. Sake lees are nutritious and functional, but are prone to spoilage. In this study, we investigated whether sake lees should be mixed with animal feed immediately or after drying. To assess the usefulness of sake lees as a poultry feed ingredient and determine the effect of sake lees on intestinal immunity, we performed a feeding trial with three treatments: a raw sake lees (RSL) diet, dried sake lees (DSL) diet, and control diet. Three-week-old broilers were fed these diets (n=8 per group) for two weeks. We then calculated feed efficiency and performed RT-qPCR to assess the effects of diet on intestinal immunity. The growth performance in the RSL diet group was equivalent to that in the control diet group. The DSL diet became difficult for broilers to eat, resulting in decreased growth performance. In the ileum of RSL-diet broilers, the mRNA expression levels of TGF-β1 and avian β-defensin (AvBD)12 were significantly increased compared to those of control diet broilers (p<0.05), and a significant correlation was observed between the two genes (p<0.05). Our results indicated that sake lees should not be dried and should be mixed immediately with feed, and this sake lees when fed to chicken activates the intestinal immunity. However, sake lees have a lower fat content than corn, and it is thus important to combine sake lees with high-energy feed.     

Role of ovary-inhibiting hormone from eyestalks of marine penaeid prawns (Parapenaeopsis hardwickii) during ovarian developmental cycle

The activity of the ovary-inhibiting hormone in the eyestalks of female prawns, Parapenaeopsis hardwickii, was determined during its ovarian developmental cycle. In the inactive (stage I) and post-spawned phases of ovarian development, the ovary-inhibiting hormone activity was found to be very high. During the phase of full vitellogenesis (stage IV), the inhibiting activity is almost negligible. After spawning (stage V) it is present, but at a lower level than during inactive and post-spawned phases. Hence it is inferred that ovary-inhibiting hormone appears to be responsible for the phase of ovarian quiescence and to control the rate of vitellogenesis.     

Experimental pathogenicity of reovirus 13p2 for juvenile American oysters Crassostrea virginica (Gmelin) and bluegill fingerlings Lepomis macrochirus (Rafinesque)

Abstract. A viral agent (13p₂), isolated from clinically normal juvenile American oysters ( Crassostrea virginica ) and characterized as a new serotype of reovirus, was tested to determine if it could replicate and produce disease in experimental juvenile oysters. Because the virus replicated well in the bluegill fry (BF-2) fish cell line, fingerling bluegills Lepomis macrochirus were included in the pathogenicity experiments. Exposure of oysters to the 13p₂ virus in ambient seawater resulted in no significant mortality and no increased virus titres or histological lesions. Virus particles were not observed in tissues of exposed oysters when examined with transmission electron microscopy (TEM). Intraperitoneal inoculation of the 13p₂ virus into bluegills resulted in 44% mortality associated with a focal necrotic hepatitis. Evidence of virus replication, when evaluated histological or by virus titration, was observed in 94% of 32 inoculated fish. Samples of infected livers examined with TEM revealed typical cytoplasmic arrays of 13p₂ virus particles in affected hepatocytes. Rising virus titres and hepatic lesions also occurred in bluegills exposed to water containing the 13p₂ virus. These results indicated the natural host of the 13p₂ virus was not the American oyster, but that it was a significant pathogen for at least one fish species.     

Gas-liquid chromatographic determination of monosodium glutamate in soups and soup bases.

A method is given for determining monosodium glutamate (MSG) in soups and soup bases by gas-liquid chromatography (GLC) of the trimethylsilyl ether derivative of glutamic acid. This method compared favorably with existing methods including GLC of the trifluoroacetate (TFA)/butyl ester derivative, and analysis on an amino acid analyzer. In addition, some samples were analyzed by GLC/mass spectrometry. The MSG content of various products ranged from approximately 0.2% in some condensed soups to 13.1% on bouillon cubes. The method, which can detect as little as 0.05%, requires only a 10 min single step derivatization at room temperature and is preferred to the TFA/butyl ester technique.