Analysis of a Long-term Experiment with Repeated-measurement Models

Long‐term experiments are a classical case of repeated measurements. Traits are measured on the same experimental unit over many years so that correlations arise between the observations made on the same plot in consecutive years. This paper describes the analysis of a three‐crop‐rotation long‐term experiment. We analysed the yields of the crops and the organic carbon content in the topsoil over 30 consecutive years. Several variance–covariance approaches are discussed and the trait‐specific best fit is interpreted. Mixed models are used to describe the structure of the experiment. Both yields and soil organic carbon show a more or less pronounced variance heterogeneity. Especially for yields, the heterogeneity of cycles and years is dominant. The consideration of correlations results in a better model fit in all cases.     

Effect of milk sampling techniques on milk composition, bacterial contamination, viability and functions of resident cells in milk.

Three different milk sampling techniques were evaluated during milk sampling: a direct aseptic collection from the udder through a sterile cannula was used as the reference technique, compared with either a manual or a mechanical sampling method. In this study 30 high-yielding Holstein-Friesian dairy cows at different stages of lactation and free of udder infection were used. For each milk sample, the influence of milk sampling techniques was determined for the following parameters: somatic cell count, milk composition, bacterial contamination, viability, in vitro phagocytosis and overall killing of Staphylococcus aureus Newbould 305, and cellular chemiluminescence. Because milk sampling occurred throughout lactation, the differences between early, mid- and late lactation were estimated. It was concluded that bacterial contamination was not significantly different in manual milking samples and the reference technique; bacterial contamination was, however, significantly (P < 0.001) higher in machine milking samples than in the reference technique. Among the different sampling techniques, no significant effects on SCC, milk composition, viability and functions of the cells isolated from milk were observed. It was found that viability, intracellular killing and cellular chemiluminescence of milk PMN were significantly lower (P < 0.05) in early lactation compared to mid-lactation. Phagocytosis was significantly (P < 0.05) higher in early lactation compared to mid- and late lactation, and no significant differences were observed between mid- and late lactation. From this study, it can be concluded that despite a higher bacterial contamination obtained with the mechanical sampling method, the 3 milk sampling techniques described in this study can be used for the evaluation of milk cell functions.     

Effects of Carbon Dioxide Exposure on Intensively Cultured Rainbow Trout Oncorhynchus mykiss: Physiological Responses and Fillet Attributes

Rainbow trout Oncorhynchus mykiss (261.6 × 24.7 g initial weight, mean × SEM) at 13.1 × 0.2 C were exposed for 94 d to one of three CO₂ treatments: control (22.1 × 2.8 mg/L), medium (34.5 × 3.8 mg/L), or high (48.7 × 4.4 mg/L). Trout were checked daily for survival, and fish were sampled at 0, 28, 56, and 84 d for physiological responses, growth, and fillet quality assessments. Trout were also challenged to a 15-min crowding stress at 93 d to assess their ability to initiate a stress response during hypercapnia. Chronically exposed trout showed nearly 100% survival through 84 d exposure (1 of 1,500 fish died). Growth and physiological results showed that increasing elevated CO₂, concentrations result in corresponding decreased growth rates and CO₂specific physiological parameters: The medium and high CO₂ treatments had significantly slower growth and subsequently smaller fish by 84 d. Exposed trout also showed significantly ( P < 0.05) decreased plasma chloride for medium and high CO₂ treatments compared to the control from 28 through 84 d. Decreased growth and smaller fish in the medium and high CO₂ treatments resulted in correspondingly smaller fresh and smoked fillet weights. Chronic CO₂ exposure did not result in notable changes in ultimate muscle pH. Exposure to 15-min crowding stress at 93 d resulted in significant changes in hematocrit, plasma cortisoI, glucose, and chloride for all treatment groups. CO₂-specific changes were detected in hematocrit, plasma cortisoI, and plasma chloride responses following the 15-min crowding stress.     

Patterns of silver eel (Anguilla anguilla L.) sex ratio in a catchment

Abstract –  Changes in the numbers and size-class structure of European silver eels, Anguilla anguilla , in the River Frémur (France) were examined over a 9-year period after installation of downstream eel passes. The number of silver eels migrating downstream peaked in 1999, then decreased strongly and steadily after 2000, reaching relatively low levels. At the same time, a gradual shift in the silver eel sex ratio from a dominance of males (size from 270 to 442 mm, age from 3 to 6 years) to females (size from 366 to 1112 mm, age from 4 to 9 years) was recorded. Possible explanations for the escapement patterns observed are environmental sex determination and the installation of eel passes on the main hydraulic engineering structures in 1992 and 1996.     

Equine Melanocytic Tumors: A Retrospective Study of 53 Horses (1988 to 1991)

A study of 57 cutaneous melanocytic tumors from 53 horses revealed 4 distinct clinical syndromes: melanocytic nevus, dermal melanoma, dermal melanomatosis, and anaplastic malignant melanoma. Melanocytic nevus and anaplastic melanoma each had histopathologic features that distinguished them from dermal melanoma and dermal melanomatosis. Dermal melanoma and dermal melanomatosis were histologically similar but could be differentiated by their clinical features. Melanocytic nevi were diagnosed in 29 horses with an average age of 5 years; they were solitary, superficial masses that occurred in both grey and nongrey horses, and in which surgical excision was generally curative. Dermal melanomas were diagnosed in 20 horses with an average age of 13 years; all horses of known coat color were grey. Eight horses with an average age of 7 years had 1 or 2 discrete dermal melanomas. Follow-up information was available for 6 horses; metastases occurred in 2 horses, and surgical excision was apparently curative in 4 horses. Dermal melanomatosis was diagnosed in 12 grey horses with an average age of 17 years; all 6 of these horses evaluated had internal metastases. In 2 aged nongrey horses with anaplastic malignant melanoma, the tumors metastasized within 1 year of diagnosis. Two tumors with features of both melanocytic nevus and dermal melanoma remained unclassified.     

Evaluation of Sperm-Activating Solutions in Atlantic Salmon Salmo salar Fertilization Tests

The use of saline solutions was tested to determine their efficacy as replacements for ovarian fluid as sperm activators and to eliminate variability encountered with the use of Ovarian fluid. We tested fertilization rate of semen from eight males on Atlantic salmon Salmo salar eggs after five sperm-activating solutions and a non-activating saline were substituted for ovarian fluid. We used solutions shown acceptable for use with other salmonid species. The six solutions tested were a non sperm-activating phosphate-buffered saline (PBS, 7.2 g/L NaCl, 1.48 g/L Na₂HPO₄, 0.43 g/L K H₂PO₄), a Tris buffer (6.99 g/L NaCl, 3.63 g/L Tris and 2.42 g/L glycine), a Borax buffer (12.2 g boric acid/L in solution 1, 76 g disodium tetraborate/L combined 100:118, then 1 L combined with 3.7 L water and 18 g NaCl), and three solutions of 9.25 g/L (125 mM) NaCl buffered to pH 6.0, 7.5, and 8.9. The latter five solutions activated sperm immediately on contact, while PBS required additional water to activate sperm. The PBS solution was the least effective (mean percent eyed eggs 37.6%) for egg fertilization. The mean percent eyed eggs for the other five saline solutions (range 78% to 86%) were not significantly different. Sperm from one male provided significantly lower egg fertilization (39.6%) when compared with the other seven males (67.2–87.4% egg fertilization). Percent egg fertilization was not related to number of live sperm cells per egg. Our results show that osmotically-balanced sperm-activation solutions, even those with a pH range from 6.0 to 8.9 provide adequate media for fertilization of Atlantic salmon eggs. Fertilization in a deactivation saline and water reactivation of sperm resulted in low egg fertilization.     

Storage of Non-structural Carbohydrates in Sweet Sorghum [Sorghum bicolor (L.) Moench.]: Comparison of Sterile and Fertile Lines

The accumulation of soluble carbohydrates in vegetative parts of sweet sorghum ( Sorghum bicolor (L.) Moench.) represents an intermediate reserve pool for grain filling as well as an irreversible storage up to maturity. In order to study the effect of Sterility on soluble sugar accumulation in vegetative parts, two cytoplasmic male sterile (CMS) lines of sweet sorghum were compared with their corresponding fertile mamtainers for non-structural carbohydrate concentrations in shoots. Plants were harvested at four different physiological stages. The lines showed significant differences in the yield of non-structural carbohydrates. At maturity, the yields of soluble carbohydrates from vegetative parts of the two sterile lines were 93 % and 43 % higher than the yield of the two corresponding fertile lines. At that stage, sucrose represented about 80 % of total soluble carbohydrates and the higher yield of soluble carbohydrates of sterile lines was mainly due to the higher yield of sucrose. This indicates that the yield of extractable carbohydrates, especially sucrose, in vegetative parts of sweet sorghum could be considerably improved by using sterile lines which are utilized in hybrid production.