Partial characterization of alkaline proteases from viscera of vermiculated sailfin catfish Pterygoplichthys disjunctivus Weber, 1991

Vermiculated sailfin catfish (Pterygoplichthys disjunctivus, Weber, 1991), a member of the Loricariidae family and an invasive species of several inland waters around the world, possess an enormous digestive tract representing about 10% of fish weight. Thus, the aim of this study was to partially characterize proteases from their digestive tracts. Azocasein digestion of the crude extract of intestine at different pH values and temperatures revealed the presence of alkaline proteases with optimum activities at pH 9.0 and 50°C. Incubation assays of the crude extract with inhibitors such as phenyl methyl sulfonyl fluoride, N-α-p-tosyl-l-lysine chloromethyl ketone, N-tosyl-phenyalanine chloromethyl ketone, benzamidine, pepstatin A and ethylenediamine tetra-acetic acid showed that trypsin and chymotrypsin are the main alkaline proteinases present. Zymography showed that the crude extract of Pterygoplichthys disjunctivus viscera contained proteases with molecular masses ranging from 21.5 to 116 kDa. Trypsin and chymotrypsin were inhibited by the following ions in decreasing order: Hg²⁺, Fe²⁺, Cu²⁺, Li⁺, Mg²⁺, K⁺, while Mn²⁺, and Ca²⁺ had no effect. Activities decreased continuously as the NaCl concentration increased from 0 to 30%. These results constitute important background information for future studies and for the potential biotechnological use of the crude digestive extract from this invasive species.     

Effects of tillage systems in dryland farming on near-surface water content during the late winter period

Tillage systems modify, at least temporarily, some of the physical properties of soil, such as soil porosity. Tillage also has an indirect effect on soil water content throughout the growth cycle, particularly in areas with a Mediterranean climate. This paper presents the results of monitoring the water content in the topsoil (0–0.20 m) of three adjacent plots during February to May cycles starting in 1994–1995 and ending in 1998–1999. Each of the plots had a surface area of 2700 m², an 8% slope and Calcic Cambisol soil. Starting in 1994, three different tillage systems were applied: conventional tillage, which is typical of the area (CT); minimum tillage (MT); and no-tillage (NT). Two vertical 200 mm TDR probes were permanently installed in each plot and measurements were taken every week. The results show that, under an NT system, the soil had significantly higher water content than the other two soil plots. However, this increased quantity of water did not denote increased crop production; on the contrary, these preliminary data point to a decrease in crop production.     

Analysis of unbalanced mixed model data: A case study comparison of ANOVA versus REML/GLS

Major transition has occurred in recent years in statistical methods for analysis of linear mixed model data from analysis of variance (ANOVA) to likelihood-based methods. Prior to the early 1990s, most applications used some version of analysis of variance because computer software was either not available or not easy to use for likelihood-based methods. ANOVA is based on ordinary least squares computations, with adoptions for mixed models. Computer programs for such methodology were plagued with technical problems of estimability, weighting, and handling missing data. Likelihood-based methods mainly use a combination of residual maximum likelihood (REML) estimation of covariance parameters and generalized least squares (GLS) estimation of mean parameters. Software for REML/GLS methods became readily available early in the 1990s, but the methodology still is not universally embraced. Although many of the computational inadequacies have been overcome, conceptual problems remain. Also, technical problems with REML/GLS have emerged, such as the need for adjustments for effects due to estimating covariance parameters. This article attempts to identify the major problems with ANOVA, describe the problems which remain with REML/GLS, and discuss new problems with REML/GLS.     

Effects of density profile of MDF on stiffness and strength of nailed joints

Nail-head pull-through, lateral nail resistance, and single shear nailed joint tests were conducted on medium density fiberboard (MDF) with different density profiles, and the relations between the results of these tests and the density profiles of MDF were investigated. The maximum load of nail-head pull-through and the maximum load of nailed joints were little affected by the density profile. However, the ultimate strength of lateral nail resistance, the stiffness, and the yield strength of nailed joints were affected by the density profile of MDF and showed high values when the surface layer of the MDF had high density. It is known that bending performance is also influenced by density profile. Therefore, the stiffness and the yield strength of nailed joints were compared with the bending performance of MDF. The stiffness of nailed joints was positively correlated with the modulus of elasticity (MOE); in the case of CN65 nails, the initial stiffness of joints changed little in response to changes in MOE. The yield strength of nailed joints had a high positive correlation with the modulus of rupture (MOR). The stiffness and the yield strength of nailed joints showed linear relationships with MOE and MOR, respectively.     

Survival analysis in two lines of rabbits selected for reproductive traits

The objective of the study was to analyze the reproductive longevity of 2 selected lines of rabbits. The first one was the Prat line, a line selected in Spain on litter size at weaning, and raised in overlapping generations. The second one was a French line, the A1077 line, selected on litter size at birth and individual weight at 63 d of age, managed in batches, and artificially inseminated with discrete generations. Reproductive longevity was measured beginning at the first successful mating, assessed by a pregnancy diagnosis in the Prat line, and at the first kindling in the A1077 line. In the A1077 line, culling for infertility occurred after 3 unsuccessful artificial inseminations. The trait analyzed, defined as the doe length of productive life (LPL), was the time in days between date of the first positive pregnancy diagnosis and date of culling or death in the Prat line. In the A1077 line, the trait was the number of AI after the first kindling. Effects included in the model were year-season, litter size at birth, reproductive cycle or physiological status x cycle interaction, age at first mating, batch (only for the A1077 line), and additive genetic value of the animal as a random effect. Survival analyses were carried out with a Cox model for the Prat line and a discrete model for the A1077 line. The estimated heritability values for LPL were around 0.16 in the Prat and A1077 lines with a model including physiological status x cycle interaction effect. Removing this effect from the model led to an increase in estimated genetic variance with h2 = 0.24 and 0.19 in the Prat and A1077 lines, respectively. Including the traits LPL and number of AI from first fertile mating or AI in selection programs could increase reproductive longevity and decrease the replacement rate.     

Crossbreeding parameter estimation for functional longevity in rabbits using survival analysis methodology

A complete diallel cross involving 3 maternal lines of rabbit was performed to estimate cross-breeding parameters for functional longevity. This trait was defined as the ability to delay involuntary culling. The lines considered, A, V, and Prat, had all been selected by litter size at weaning for a long period. Data were related to a total of 653 does belonging to the 9 genetic types from the diallel cross; does were reared and bred on the same commercial farm. Survival analysis was performed using a Cox proportional hazard model. The model incorporated time-dependent factors, such as year-season, litter size, and the interaction between cycle and physiological status of the female; time-independent factors, such as the genetic type of the doe; and sire and dam random factors. Crossbreeding parameters were estimated from the solutions obtained for the type of doe and its estimated variance-covariance matrix, using a generalized least squares procedure. The estimated parameters were the differences between lines in direct genetic effects and maternal genetic effects and individual heterosis. Relevant differences were observed in direct genetic effects between lines A and Prat but not in any maternal effects. Heterosis was found to be significant and favorable between lines A and Prat, and between the lines V and Prat. The magnitude of this effect was variable but important, especially in the first cross. Results stress the importance of using crosses between specialized lines to produce does for intensive meat rabbit production.     

Genetic basis of semen traits and their relationship with growth rate in rabbits

This work aims to estimate the genetic parameters of seminal and production traits in a paternal line of rabbits selected for ADG during the fattening period. The considered traits were male libido (Lib) defined as successful mounting of an artificial vagina; presence of urine (Ur) and calcium carbonate deposits (Ca) in the ejaculate; semen pH; individual sperm motility (IM); the suitability for AI of the ejaculate (Sui), which involves the subjective combination of several quality traits; the average ejaculate volume (Vol); sperm concentration (Conc); and the average sperm production per ejaculate (Prod = Vol × Conc). The genetic relationship between all of these traits with ADG is also provided. Male libido and seminal data came either from routine evaluations of the ejaculates in an AI center or from 2 experiments in which bucks from the same population were used. Two consecutive ejaculates per male and per week were collected, leaving 7 d within weekly collections. A linear tri-trait model was used to analyze Conc, Vol, and ADG, whereas linear and threshold-linear 2-trait models were used to analyze male libido and the remaining seminal traits with ADG. A Bayesian approach was adopted for inference. Approximately 38% of ejaculates were rejected for AI primarily due to low IM scores. Variables related to the quality of the ejaculate (Ur, Ca, pH, IM, Sui) and Lib were found to be lowly heritable (h(2) ranged from 0.04 to 0.11), but repeatable. This indicates performance of bucks for seminal quality traits and libido in AI centers would be more strongly affected by management practices rather than genetic selection. Semen production traits exhibited moderate values of h(2) (0.22, 0.27, and 0.23 for Conc, Vol, and Prod, respectively), suggesting the possibility of effective selection for these traits. A moderate to high negative genetic correlation (r(g); posterior mean; highest posterior density at 95%, HPD(95%)) was estimated between Conc and Vol (-0.53, HPD(95%) = -0.76, -0.27). The ADG was estimated to have an h(2) of 0.16, to have a low, positive r(g) with Conc (0.21, HPD(95%) = -0.03, 0.48), to have a low, negative r(g) with Vol (-0.19, HPD(95%) = -0.47, 0.08), and to be genetically uncorrelated with all remaining traits analyzed. Therefore, selection for increasing ADG in paternal lines is expected to have no detrimental effects on Ur, Ca, pH, IM, Sui, and Lib and little to no effect on Conc, Vol, and Prod.     

Evaluation of the efficacy of commercial vaccines against bluetongue virus serotypes 1 and 8 in experimentally infected red deer (Cervus elaphus)

Red deer (Cervus elaphus) is a widespread and abundant species susceptible to bluetongue virus (BTV) infection. Inclusion of red deer vaccination among BTV control measures should be considered. Four out of twelve BTV antibody negative deer were vaccinated against serotype 1 (BTV-1), and four against serotype 8 (BTV-8). The remaining four deer acted as unvaccinated controls. Forty-two days after vaccination (dpv), all deer were inoculated with a low cell passage of the corresponding BTV strains. Serological and virological responses were analyzed from vaccination until 28 days after inoculation (dpi). The vaccinated deer reached statistically significant (P<0.05) higher specific antibody levels than the non vaccinated deer from 34 (BTV-8) and 42 (BTV-1) dpv, maintaining stable neutralizing antibodies until 28 dpi. The non vaccinated deer remained seronegative until challenge, showing neutralizing antibodies from 7 dpi. BTV RNA was detected in the blood of the non vaccinated deer from 2 to 28 dpi, whereas no BTV RNA was found in the vaccinated deer. BTV was isolated from the blood of non vaccinated deer from 7 to 28 dpi (BTV-1) and from 9 to 11 dpi (BTV-8). BTV RNA could be identified by RT-PCR at 28 dpi in spleen and lymph nodes, but BTV could not be isolated from these samples. BT-compatible clinical signs were inapparent and no gross lesions were found at necropsy. The results obtained in the present study confirm that monovalent BTV-1 and BTV-8 vaccines are safe and effective to prevent BTV infection in red deer. This finding indicates that vaccination programs on farmed or translocated red deer could be a useful tool to control BTV.     

Social behaviour of buffalo heifers during the establishment of a dominance hierarchy

This study investigated the social behaviour of buffalo heifers during the establishment of a dominance hierarchy under conditions that differed in space availability (pen, small pasture and large pasture), food competition and in the presence or absence of an intact bull. The herd consisted of 21 Mediterranean–Murrah crossbreed buffalo heifers and one bull. During the establishment of dominance hierarchy, one observer recorded all of agonistic interactions occurring in the pen, in the small pasture and in the large pasture. For the food competition test, the females were first separated into groups (G) of seven animals and the behavioural interactions within these groups were observed. The group levels were determined by the sociometric parameters (h′: Landau linearity index and DCI: directional consistency index). The percentage of unknown, one-way and two-way dyadic relationships; a social index, hierarchical position and correlations among feeding duration, latency, social index and hierarchical position were calculated. In the large pasture, the DCI value was 55.24% of the dyadic relationships were one-way and 36.19% were unknown; whereas in the small pasture and pen most of the dyads were unknown (54–63%) or one-way (33-41%). The social indexes observed were low (0.004 and 0.122). In the food competition test, h′ values were not significant for groups G1, G2, G3 and G6, and significant for Groups G4 and G5. The percentages of two-way dyadic relationships were high in G4 (80.95%) and G6 (100%) groups. Correlations among feeding duration and latency, social position and social index were all negative and significant. There were no significant correlations between body weight and social index or hierarchy position. This study showed that buffalo heifers established social dominance relationships among several of the herd members, but because there were several unknown dyads, the hierarchy is classified as semi-linear hierarchy. Two factors seemed to affect the establishment of social dominance and a hierarchy: adjusting to a new environment and the presence of an adult intact male, and the dominant females had advantage over the subordinate females during feeding in a trough. To reduce agonistic encounters between buffalo heifers suggest provide more space in the trough during feeding.