Effect of Relaxin on Fertility Parameters of Frozen–Thawed Buffalo (Bubalus bubalis) Sperm

The aim of this work was to evaluate the effect of relaxin on fertility parameters of buffalo frozen/thawed sperm. Sperm were incubated in the absence of capacitating agents (negative control), with a known capacitating agent such as heparin (positive control) and with 50 and 100 ng/ml relaxin for 2 and 4 h. Sperm viability, motility, capacitation and the effect of relaxin on the fertilizing ability after heterologous IVF were evaluated. Although viability was not affected, relaxin increased (p < 0.05) sperm motility compared to the negative and positive controls both after 2 h (60.0 ± 2.0, 60.0 ± 3.1, 68.3 ± 1.7 and 69.4 ± 2.7, respectively, in negative control, positive control, 50 and 100 ng/ml relaxin) and 4 h (55.0 ± 2.5, 53.3 ± 3.0, 62.2 ± 3.0 and 65.0 ± 3.2, respectively, in negative control, positive control, 50 and 100 ng/ml relaxin) incubation. When sperm were incubated with both 100 ng/ml relaxin and heparin, a decrease (p < 0.01) of pattern A, that is low capacitation level, was observed compared to the negative control both after 2 h (54.4, 34.3 and 36.4%, respectively, in negative control, positive control and 100 ng/ml relaxin) and 4 h (51.9, 35.0 and 34.3%, respectively, in negative control, positive control and 100 ng/ml relaxin). Moreover, an increase (p < 0.01) of pattern EA, that is high capacitation level, was recorded with 100 ng/ml relaxin and heparin compared to the negative control both after 2 h (44.1, 59.3 and 57.7%, respectively, in negative control, positive control and 100 ng/ml relaxin) and after 4 h (43.0, 54.4 and 56.0%, respectively, in negative control, positive control and 100 ng/ml relaxin). Finally, relaxin increased (p < 0.01) cleavage rate compared to the negative control (57.1 ± 4.4, 72.5 ± 6.0, 71.4 ± 5.5 and 73.6 ± 2.9, respectively, in negative control, positive control, 50 and 100 ng/ml relaxin). In conclusion, relaxin has a beneficial effect on motility, capacitation and fertilizing ability of frozen–thawed buffalo sperm.     

Effect of protein supplementation on expression and distribution of urea transporter-B in lambs fed low-quality forage

Two experiments were conducted to determine the effects of ruminal protein degradability, supplementation frequency, and increasing dietary protein on the expression and distribution of urea transporter-B (UT-B) in lambs fed low-quality forage (mature crested wheatgrass hay; 4.2 to 4.7% CP). In Exp. 1, 15 Dorset wether lambs (initial BW=45.8+/-1.3 kg) were blocked by initial BW and assigned to 1 of 3 treatments within a randomized complete block design for 28 d, with supplements fed to achieve 7, 10, or 13% total dietary CP. In Exp. 2, 13 Dorset wether lambs (initial BW=34+/-4 kg) were used in a completely randomized design and given 1 of 4 isonitrogenous supplements: 1) ruminally degradable protein (RDP) fed daily (n=3), 2) RDP fed on alternate days (n=3), 3) ruminally undegradable protein (RUP) fed on alternate days (n=3), or 4) a 50:50 mixture of RDP and RUP fed on alternate days (n=4) for 18 d. Alternate-day treatments were fed at twice that of daily supplementation. On the last day of both experiments, lambs were killed and samples taken for Western blot analyses for UT-B. Immunoblotting using a rabbit polyclonal antibody to UT-B confirmed the presence of distinct 32-kDa (consistent with a nonglycosylated UT-B protein) and 47-kDa (probable N-glycosylated form of UT-B) protein bands in all 9 tissues analyzed. In both experiments, the liver, dorsal rumen, reticulum, and ventral rumen displayed strong bands at 32 kDa and lighter bands at 47 kDa, whereas the cecum, large colon, spiral colon, and parotid salivary gland displayed slight 32-kDa bands and stronger, more visible bands at 47 kDa. Both protein bands were apparent in the kidney at similar visual intensities in Exp. 1, whereas the relative intensities of the 2 UT-B bands in the kidney were variable, and appeared somewhat reciprocal among animals in Exp. 2. Although the abundance of the 47-kDa UT-B band in the ventral rumen was greater (P=0.03) in lambs fed RDP daily in Exp. 2, no other treatment differences (P >or= 0.15 to 0.99) in the abundance of the 32- or 47-kDa UT-B proteins within tissues were observed in either experiment. Although protein supplementation strategy had little effect on UT-B expression in tissues other than the ventral rumen, differences in the degree of glycosylation of UT-B across tissues may provide insight into its regulation.     

Vigor-controlling rootstocks affect early shoot growth and leaf area development of kiwifruit

Patterns of shoot development and the production of different types of shoots were compared with scion leaf area index (LAI) to identify how eight clonal Actinidia rootstocks influence scion development. Rootstocks selected from seven Actinidia species (A. chrysantha Merri., A. deliciosa (A. Chev.) C. F. Liang et A.R. Ferguson, A. eriantha Benth., A. hemsleyana Dunn, A. kolomikta (Maxim. et Rupr.) Maxim., A. kolomikta C.F. Liang and A. polygama (Sieb. et Zucc.) Maxim.) were grafted with the scion Actinidia chinensis Planch. var. chinensis 'Hort16A' (yellow kiwifruit). Based on an earlier architectural analysis of A. chinensis, axillary shoot types produced by the scion were classified as short, medium or long. Short and medium shoots produced a restricted number of preformed leaves before the shoot apex ceased growth and aborted, resulting in a 'terminated' shoot. The apex of long shoots continued growth and produced more nodes throughout the growing seasons. Mid-season LAI of the scion was related to the proportion of shoots that ceased growth early in the season. Scions on low-vigor rootstocks had 50% or less leaf area than scions on the most vigorous rootstocks and had a higher proportion of short and medium shoots. On low-vigor rootstocks, a higher proportion of short shoots was retained during pruning to form the parent structure of the following year. Short parent shoots produced a higher proportion of short daughter shoots than long parent shoots, thus reinforcing the effect of the low-vigor rootstocks. However, overall effects of rootstock on shoot development were consistent regardless of parent shoot type and nodal position within the parent shoot. Slower-growing shoots were more likely to terminate and scions on low-vigor rootstocks produced a higher proportion of slow-growing shoots. Shoot termination also occurred earlier on low-vigor rootstocks. The slower growth of terminating shoots was detectable from about 20 days after bud burst. Removal of a proportion of shoots at the end of bud burst increased the growth rate and decreased the frequency of termination of the remaining shoots on all rootstocks, indicating that the fate of a shoot was linked to competitive interactions among shoots during initial growth immediately after bud burst. Rootstock influenced the process of shoot termination independently of its effect on final leaf size. Scions on low-vigor rootstocks had a higher proportion of short shoots and short shoots on all rootstocks had smaller final leaf sizes at equivalent nodes than medium or long shoots. Only later in the development of long shoots was final leaf size directly related to rootstock, with smaller leaves on low-vigor rootstocks. Thus, the most important effect of these Actinidia rootstocks on scion development occurred during the initial period of shoot growth immediately after bud burst.     

Effects of increasing days of exposure to prepartum diets on the concentration of certain blood metabolites in dairy cows

Objective To evaluate the effect of the number of days exposure to a prepartum transition diet on mineral, energy and protein metabolism in dairy cows. Design A prospective cohort study. Procedures The prepartum transition diet consisted of ryegrass pasture, cereal hay, grain, grain by-product, protein meals, BioChlor®, rumen modifiers, minerals and vitamins and contained 13.9% crude protein, 10.0 MJ metabolisable energy/kg and a dietary cation anion difference of −35 meq/kg dry matter. Forty cows were bled bi-weekly from their introduction to the prepartum transition diet until day 35 of lactation. Blood samples were submitted for estimation of a range of metabolites. Cubic smoothed splines were fitted to scatterplots of metabolite concentration as a function of day relative to calving and the area under the curve (AUC) was calculated. Linear regression modelling determined the effect of days exposed, age, body condition score (BSC) and calving day on AUC. Results The prepartum AUC of blood phosphorus and beta-hydroxybutyrate increased and the AUC of blood calcium and cholesterol decreased with increasing days exposed to the diet. The postpartum AUC of beta-hydroxybutyrate and non-esterified fatty acids decreased with increasing days exposed but increased with days exposed × age and days exposed × BCS, respectively. The AUC of other metabolites did not vary significantly with number of days exposed. Conclusions Increasing exposure to the prepartum diet significantly altered the AUC of blood metabolites associated with mineral, energy and protein metabolism in ways consistent with improvements in production and reproduction previously reported and the result suggests links between energy, protein and skeletal metabolism.     

Vibrios as causal agents of zoonoses

Vibrios are Gram-negative rod-shaped bacteria that are widespread in the coastal and estuarine environments. Some species, e.g. Vibrio anguillarum and V. tapetis, comprise serious pathogens of aquatic vertebrates or invertebrates. Other groups, including Grimontia (=Vibrio) hollisae, Photobacterium (=Vibrio) damselae subsp. damselae, V. alginolyticus, V. harveyi (=V. carchariae), V. cholerae, V. fluvialis, V. furnissii, V. metschnikovii, V. mimicus, V. parahaemolyticus and V. vulnificus, may cause disease in both aquatic animals and humans. The human outbreaks, although low in number, typically involve wound infections and gastro-intestinal disease often with watery diarrhoea. In a minority of cases, for example V. vulnificus, there is good evidence to actually associate human infections with diseased animals. In other cases, the link is certainly feasible but hard evidence is mostly lacking.     

Variation in juvenile steelhead (Oncorhynchus mykiss) dispersal in relation to age and growth performance

Volitional dispersal is a ubiquitous strategy characteristic of species across major faunal groups. Dispersal during the juvenile life stage is of interest because early performance can be critical for determining future success (survival/reproduction). For salmonids, dispersal can influence local density, competition, individual growth and survival, though drivers of dispersal at meso-scales are rarely quantified. Here, we evaluate dispersal of tagged juvenile steelhead (Oncorhynchus mykiss) through habitat units in extended stream reaches (500 m) at sites across a watershed from July to October 2017. Our aim was to quantify the frequency and spatial extent of dispersal, identify links to biotic and abiotic factors, evaluate the implications for individual growth and test for associations between dispersal and migration initiation. Dispersal rates varied between sites, but were consistently higher for age 1+ than for age 0 steelhead (avg. 21% vs. 6% respectively). Age 1+ dispersal probability was positively correlated with time between recapture events and body mass, and negatively correlated with growth rate, maximum temperature experienced and age 1+ density. At sites where there appeared to be growth benefits to remaining sedentary compared to moving, proportionally fewer fish performed dispersal. We found no links between dispersal and timing or probability of migration initiation the following spring. Our results support the hypothesis that although dispersal over intermediate scales (10–1,000 m) might be rare, it could be an important strategy that permits fish to seek out better opportunities (foraging, shelter or otherwise) in underutilised areas.     

Effects of Clinical Mastitis on Ovarian Function in Post-partum Dairy Cows

Mastitis-induced ovarian abnormalities were studied in a field trial. At 1-3 day after calving, > or = 2 parity cows not affected with chronic recurrent mastitis and yielding < 400,000/ml somatic cell count (SCC) individual milk in the previous lactation, were enrolled in the study. Thereafter milk samples were collected three times weekly for 95-100 day for progesterone (P4) assay. Individual P4 profiles were used to monitor ovarian cyclicity. When mastitis was diagnosed in the first 80 day post-partum (pp), clinical signs were recorded and scored, and aseptic milk samples were taken to identify the mastitis pathogens. Depending on the isolated pathogens the cows were blocked into one of the three sub-groups affected by either Gram-positive (GP), or Gram-negative (GN) bacteria, or of those with no detected pathogens (NDP). Cows suffering from any type of mastitis between days 15 and 28 (n = 27) showed a delay in the onset of ovarian cyclicity, and estrus was postponed compared to cows affected during the first 14 day pp (n = 59) and controls (n = 175) (38.6 +/- 2.3 vs 33.4 +/- 2.1 and 32.0 +/- 1.0 day, respectively, for onset of ovarian cyclicity and 90.7 +/- 2.5 vs 80.2 +/- 2.8 and 83.9 +/- 2.1 day, respectively, for estrus; both p < 0.05). The percentage of cows ovulating by day 28 was lower in those affected by mastitis between days 14 and 28 compared to cows between days 1 and 14 and controls (22.2% vs 47.5 and 50.3%, respectively; p < 0.05). A significantly higher rate of premature luteolysis was observed in GN + NDP compared to GP mastitis and healthy cows (46.7% vs 8.3 and 2.0%, respectively; p < 0.001). If the mastitis outbreak occurred during the follicular phase, the duration of this cycle segment was lengthened in GN + NDP mastitis compared to GP mastitis and healthy cows (10.8 +/- 0.9 vs 7.9 +/- 0.1 and 7.2 +/- 0.1, respectively; p < 0.001). The results indicate that mastitis can affect the resumption of ovarian activity in pp dairy cows. Mastitis may also impair reproduction also in cyclic cows: this effect can be the consequence of premature luteolysis or a prolonged follicular phase.