Effects of Sperm Concentration and Straw Volume on Motion Characteristics and Plasma, Acrosomal, and Mitochondrial Membranes of Equine Cryopreserved Spermatozoa

The advantages of using cryopreserved semen in equine reproduction are well known. During cryopreservation, spermatozoa undergo many changes that lead to a decrease in fertility. There is no agreement on the ideal sperm dose and concentration to maximize fertility rates. Thus, the objectives of this experiment were to evaluate sperm motion by computer-assisted analysis (CASA), sperm membrane integrity and function with fluorescence probes of cryopreserved sperm at three concentrations: 100 (C100), 200 (C200) and 400 × 10⁶ sperm/mL (C400), and two straw volumes (0.50 and 0.25 mL). There was no interaction between sperm concentration and storage volume (P > .05). Sperm motion characteristics were influenced by concentration (C100 > C200 > C400; P < .05). Curvilinear velocity (VCL) in 0.25-mL straws had higher average values (P < .05). Membrane integrity and function were not changed by straw volume (P > .05). However, sperm concentration changed the percentage of cells with intact plasma membrane (C100 > C200 > C400; P < .05) and the percentage of cells with high mitochondrial membrane potential (C100 = C200; P > .05 and C400 < C100 and C200; P < .05). According to this experiment, the best freezing method was that involving 100 × 10⁶ sperm/mL, regardless of straw volume.     

The persistence of foot-and-mouth disease virus on wool

SUMMARY Five Suffolk sheep, held in a high-security isolation room, were exposed for 2 hours to the aerosol of 3 mature pigs that had been infected with foot-and-mouth disease virus (FMDV), strain O₁-BFS. The fleeces of 3 of the sheep were contaminated with FMDV at 2 days post exposure (dpe), while at 5 dpe the fleeces of all 5 sheep were more extensively, and more heavily, contaminated. The persistence of FMDV on contaminated wool was examined in vitro using multiple 0.5 g samples of Merino wool that were each contaminated with one of 3 strains of FMDV in tissue-culture medium: O₁-BFS, O-Morocco (O-MOR 9/91) or an Asia 1 strain (TAI 1/90). Wool samples were held at either 4°C, 18°C or 37°C, and decay curves were established for each virus at each temperature. These curves predicted that O₁-BFS, O-MOR 9/91 and TAI 1/90 would fall below detect-able levels at 72, 70 and 48 days post contamination (pc), respectively, for wool stored at 4°C; at 11, 12 and 12 days pc, respectively, for wool stored at 18°C; and at 57, 68 and 33 hours pc, respectively, for wool stored at 37°C. For wool contaminated with O₁-BFS-infected sheep faeces, urine or blood, or with O₁-BFS-infected cattle saliva, decay curves predicted virus to persist for 5 to 11 days pc at 18°C. We demonstrated that the simulated scouring of FMDV-contaminated wool at 60° to 70°C would usually reduce virus to below detectable levels. The detergent component of the scouring process had little, if any, antiviral activity, and scouring at 20°C or 50°C had limited impact on FMDV titres . We recommend that either (1) simple storage of FMDV-contaminated wool for 4 weeks at temperatures of 18°C or higher, or (2) scouring of contaminated wool at 60° to 70°C would be sufficient to remove the threat of FMDV-contaminated wool being infectious to other animals .     

The impact of sugarcane filter cake on the availability of P in the rhizosphere and associated microbial community structure

This study aimed to evaluate the interaction between filter cake (FC), and phosphorus fertilizers with differing solubility on the growth and P nutrition of sugarcane. Effects of soil amendment with FC on different soil P fractions and influence on microbial community structure in the rhizosphere were also assessed. Two glasshouse experiments were conducted with completely randomized block designs. The first experiment evaluated rates of FC using a factorial design (5 × 2): 0, 2.5, 5, 10 and 15 g FC kg^?1 soil applied as either broadcast in bulk soil or in the planting furrow. The second experiment used a factorial design (2 × 3): without and with FC (5 g kg^?1 soil, dry basis), both without P (NP) and with P supplied as either triple superphosphate (TSP) or as rock phosphate (RP), both at the rate of 78.4 mg kg^?1 based on total P. Microbial community structure was determined using TRFLP and dynamics of soil P by Hedley fractionation. Filter cake applied at increasing rates in the absence of P was effective in increasing shoot growth and P uptake by plant, particularly when applied to bulk soil as compared to furrow application. Also, FC improved P uptake and increased the availability of labile inorganic P in the rhizosphere and modified the structure of fungal and bacterial communities, whereas only bacterial and archaea communities were influenced by P fertilizer use. Filter cake was more effective when combined with RP, with increased growth and P utilization and thus can be considered as a feasible and practical option for farmer use in substitution to TSP, a more expensive source.     

Antiproliferative effect of Canavalia brasiliensis lectin on B16F10 cells

Lectins are proteins or glycoproteins with the ability to link to carbohydrates at the cell surface in a specific and reversible manner. Studies have showed that lectins are demonstrate immunomodulatory and antitumor agents. This study aims to evaluate the effect of lectin extracted from the seeds of Canavalia brasiliensis (ConBr) on murine melanoma B16F10 cells by analyzing cell viability, apoptosis index, cell migration, production of cytokines and nitric oxide (NO). Results showed that ConBr was able to reduce cell viability and thwart apoptosis, which could be observed by decrease in cell migration. ConBr also induced NO and IL-12 synthesis. Altogether, these data demonstrate the potential of ConBr as a therapeutic agent for melanoma.     

Cessation of clinical disease and spirochete shedding after tiamulin treatment in pigs experimentally infected with “Brachyspira hampsonii”

With the emergence of “Brachyspira hampsonii” associated with swine dysentery in North America, identification of effective treatments and interventions is a pressing need. Denagard® (tiamulin hydrogen fumarate) Liquid Concentrate 12.5% is approved in the United States for treatment of dysentery caused by Brachyspira hyodysenteriae at 0.006% in the water. In this study, the effectiveness of tiamulin in resolving clinical disease, eliminating viable spirochete shedding, and reducing neutrophilic colitis following infection with either “B. hampsonii” or B. hyodysenteriae was evaluated. Seventy-eight 7-week-old crossbred pigs were divided into three groups [sham-inoculated (n = 18), “B. hampsonii”-inoculated (n = 30), and B. hyodysenteriae-inoculated (n = 30)]. Each inoculum group was divided into three subgroups which received either 0.006% tiamulin, 0.018% tiamulin, or no medication. Both levels of tiamulin resolved clinical disease within 24 h of treatment initiation, eliminated spirochete shedding within 72 h of treatment initiation, and resolved and/or prevented histologic lesions in pigs infected with either Brachyspira spp.     

Use of Cholesterol‐Loaded Cyclodextrin in Donkey Semen Cryopreservation Improves Sperm Viability but Results in Low Fertility in Mares

The use of cholesterol‐loaded cyclodextrin (CLC) on semen cryopreservation has been related with better sperm viability in several species; however, the effect on fertility is not known in donkey semen. Ejaculates (n = 25) from five donkeys were diluted in S‐MEDIUM with 0, 1, 2 or 3 mg of CLC/120 × 10⁶ spermatozoa. Semen was frozen, and thawed samples were evaluated by computer‐assisted sperm analyser system (CASA), supravital test, hyposmotic swelling test and fluorescent dyes to assess the integrity of sperm membranes. Mares (n = 60) were inseminated with frozen‐thawed semen treated with the doses of 0 or 1 mg CLC. Percentages of sperm with progressive motility and with functional plasma membrane were greater (p < 0.05) in the CLC‐treated groups than in the control. Percentages of intact plasma membrane and intact plasma membrane and acrosome detected by fluorescent dyes were also greater (p < 0.05) in CLC‐treated groups. Although no difference (p > 0.05) in conception rates was detected between groups (control, 3/30, 10%; CLC‐treated, 1/30, 3.3%), fertility was low for artificial insemination programs in mares. Therefore, we firstly demonstrated that frozen semen treated with CLC in S‐MEDIA extender before freezing improves the in vitro sperm viability, but semen treated or not with CLC in S‐MEDIUM extender results in a very low conception rate in mares inseminated with thawed donkey semen.     

Influence of seminal plasma during different stages of bovine sperm cryopreservation

This study aimed to evaluate the effect of seminal plasma on bovine sperm cryopreservation and to assess the integrity of plasma and acrosomal membranes, mitochondrial potential, remodelling of F-actin cytoskeleton and sperm chromatin fragmentation during the cooling, equilibrium and freezing/thawing stages. Six ejaculates collected from seven Nelore bulls (n = 42) were used in this study. Each ejaculate was divided into two aliquots (with seminal plasma = SP group; without seminal plasma = NSP group) and packed to a final concentration of 50 × 10⁶ sperm per straw. Statistical analyses were performed using SAS software (version 9.3), and p ≤ .05 was considered significant. A time effect was observed for all sperm characteristics (p < .05), except for chromatin fragmentation (p > .05). The presence of seminal plasma better preserved the acrosomal integrity (SP = 75.2% and NSP = 71.7%; p < .05) and also provided lower F-actin remodelling during cryopreservation process (SP = 29.9% and NSP = 32.4%; p < .05). Regarding to the cryopreservation stages, it was observed that cooling step induced higher remodelling of F-actin than the equilibrium and freezing/thawing stages (56.3%, 32.2% and 23.9%, respectively; p < .05). The equilibrium step had minor influence on overall sperm characteristics while the freezing/thawing stage was responsible for the highest percentage of damage in plasma membrane (−65.2%), acrosomal membrane (−34.0%) and mitochondrial potential (−48.1%). On the other hand, none of the cryopreservation stages affected chromatin integrity. It was concluded that the presence of seminal plasma provides increased acrosomal integrity and reduced remodelling of F-actin cytoskeleton. Higher F-actin remodelling is observed after the cooling step while the freezing/thawing step is most damaging to sperm membranes and mitochondrial potential during bovine sperm cryopreservation.     

Visualization and application of disease diagnosis codes for population health management using porcine diseases as a model

Accurate and timely results of diagnostic investigations and laboratory testing guide clinical interventions for the continuous improvement of animal health and welfare. Infectious diseases can severely limit the health, welfare, and productivity of populations of animals. Livestock veterinarians submit thousands of samples daily to veterinary diagnostic laboratories (VDLs) for disease diagnosis, pathogen monitoring, and surveillance. Individual diagnostic laboratory reports are immediately useful; however, aggregated historical laboratory data are increasingly valued by clinicians and decision-makers to identify changes in the health status of various animal populations over time and geographical space. The value of this historical information is enhanced by visualization of trends of agent detection, disease diagnosis, or both, which helps focus time and resources on the most significant pathogens and fosters more effective communication between livestock producers, veterinarians, and VDL professionals. Advances in data visualization tools allow quick, efficient, and often real-time scanning and analysis of databases to inform, guide, and modify animal health intervention algorithms. Value is derived at the farm, production system, or regional level. Visualization tools allow client-specific analyses, benchmarking, formulation of research questions, and monitoring the effects of disease management and precision farming practices. We present here the approach taken to visualize trends of disease occurrence using porcine disease diagnostic code data for the period 2010 to 2019. Our semi-automatic standardized creation of a visualization platform allowed the transformation of diagnostic report data into aggregated information to visualize and monitor disease diagnosis.     

For how long does the quality and quantity of residues in the soil affect the carbon compartments and CO<Subscript>2</Subscript>-C emissions?

Purpose

The mineralization/immobilization of nutrients from the crop residues is correlated with the quality of the plant material and carbon compartments in the recalcitrant and labile soil fractions. The objective of this study was to correlate the quality and quantity of crop residues incubated in the soil with carbon compartments and CO₂-C emission, using multivariate analysis.

Materials and methods

The experiment was conducted in factorial 4?+?2?+?5 with three replicates, referring to three types of residues (control, sugarcane, Brachiaria, and soybean), and two contributions of the crop residues in constant rate, CR (10 Mg ha^?1 residue), and agronomic rate, AR (20, 8, and 5 Mg ha^?1 residue, respectively, for sugarcane, soybean, and Brachiaria), evaluated five times (1, 3, 6, 12, and 48 days after incubation). At each time, we determined the CO₂-C emission, nitrogen and organic carbon in the soil, and the residues. In addition, the microbial biomass and water-soluble, labile, and humic substance carbons fractionated into fulvic acids, humic acids and humin were quantified.

Results and discussion

Higher CO₂-C emissions occurred in the soil with added residue ranging from 0.5 to 1.1 g CO₂-C m^?2 h^?1 in the first 6 days of incubation, and there was a positive correlation with the less labile organic soil fractions as well as residue type. In the final period, after 12 days of soil incubation, there was a higher relation of CO₂-C emission with carbon humin. The sugarcane and soybean residue (20 Mg ha^?1) promoted higher CO₂-C emission and the reduction of carbon residue. The addition of residue contributed to an 82.32 % increase in the emission of CO₂-C, being more significant in the residue with higher nitrogen availability.

Conclusions

This study shows that the quality and quantity of residue added to soil affects the carbon sequestration and CO₂-C emission. In the first 6 days of incubation, there was a higher CO₂-C emission ratio which correlates with the less stable soil carbon compartments as well as residue. In the final period of incubation, there is no effect of quality and quantity of residue added to soil on the CO₂-C emission.

     

Daily activity pattern of the marine shrimp Litopenaeus vannamei (Boone 1931) juveniles under laboratory conditions

Lack of information on the daily activity patterns of Litopenaeus vannamei (Boone 1931) might result in inadequate feed management strategies in shrimp farms. In order to provide information to improve feed management in shrimp farms, the daily activity of juvenile shrimp reared in glass aquaria was studied, using L. vannamei (7.57±1.01 g), equivalent to 33 shrimp m^?2. Thirty‐two shrimp were observed, half in light and half in dark phase, over 280 h in 15 min h^?1 windows. Feed was provided once a day, at 10% of tank biomass. Swimming, exploration of substrate, inactivity and cleaning were recorded through instantaneous focal sampling. During the dark phase, swimming was predominant and alternated with exploration. In the light phase, inactivity was predominant, and again alternated with exploration. Cleaning occurred evenly in both the light and the dark. Substrate exploration, which characterizes the search for food, occurred in both dark and light phases, with a most intense peak 7 h after light phase onset, indicating this as the optimal feed offer.