Cryopreservation effects on the sperm quality of cachama blanca Piaractus brachypomus (Cuvier 1818)

The effects of straws volume, cryoprotectants and thawing temperatures were evaluated on the sperm quality of cachama blanca Piaractus brachypomus (Cuvier), an important Colombian fish species. Sexually mature fish were induced to ovulation or spermiation with a carp pituitary extract. A pool of suitable sperm samples was diluted in glucose, egg yolk, dimethyl sulphoxide (DMSO‐10%), methanol (MET‐10%) or ethylene glycol (ETG‐5%) and packed in 0.5, 2.5 or 5.0 mL straws and frozen in nitrogen vapour. The thawing process was performed in a 35 or an 80 °C water bath. The fertility was evaluated after 6 h post fertilization. The highest motility percentage (33 ± 3%) was observed with sperm cryopreserved with DMSO, packed in 5 mL straws and thawed at 35 °C. The treatments with DMSO and MET packed in 0.5 and 5.0 mL straws and thawed at 35 °C showed the highest fertility (higher than 71%) and the lowest fertility was obtained with MET‐2.5 mL (9 ± 5%). In all the treatments, a significant decrease in the sperm quality was observed at 80 °C. Sperm cryopreserved with DMSO‐10% or MET‐10%, packed in 2.5 or 5.0 mL straws are suitable to achieve acceptable fertilization and to fertilize high amounts of eggs.     

Effects of an Enterococcus faecium‐based probiotic on growth performance and health of Pirarucu,Arapaima gigas

Intensive fish farming has resulted in an increased concern for disease outbreaks. Probiotic use is one of the strategies being developed to improve fish health and productivity. Measures of probiotic colonization, growth performance, haematological characteristics and parasite load were used to evaluate the effect of diets supplemented with Enterococcus faecium on growth and health of Arapaima gigas juveniles. A completely randomized design with four treatments (diet with E. faecium at 1 × 10⁶ CFU/g and 1 × 10⁸ CFU/g, control diet and diet with the culture medium MRS) and three replicates was used. Ninety‐six Arapaima juveniles were distributed in 12 cages fed with the specified diet for 68 days. Colonization of the intestinal tract by lactic acid bacteria reduced the total number of heterotrophic bacteria in fish fed with probiotics compared to controls. Fish fed a supplemented diet containing 1 × 10⁸ CFU/g presented higher values of weight gain, survival and fish growth uniformity, and lower values of feed conversion ratio. The prevalence of Trichodina sp. could have affected the survival of fish in the control group. Reduction in parasite load and an increase in haematocrit, the number of erythrocytes, thrombocytes, neutrophils and monocytes were also observed in fish fed the diet containing 1 × 10⁸ CFU/g. Enterococcus faecium presented a probiotic effect in A. gigas juveniles and can be recommended for use at a concentration of 1 × 10⁸ CFU/g to modify the gut microbiota, improve growth performance and haematology and reduce parasitic load.     

Duration of the persistent activity of moxidectin against Haemonchus contortus in sheep

SUMMARY Weaned lambs were infected with Haemonchus contortus 35,28,21,14 and 7 days after treatment with moxidectin at 0.2 mg/kg and 35 and 14 days after treatment with ivermectin at the same dose rate. Worm counts 14 days after infection showed that moxidectin prevented the establishment of over 99% of infective larvae for 28 days and reduced the establishment rate at 35 days by 96%, relative to ivermectin. There was no difference in the protective efficacy of ivermectin at 14 or 35 days. The persistence of moxidectin is likely to provide advantages in nematode control, particularly when used as a strategic early summer treatment or as a pre-lambing treatment to ewes. Implications of the persistent activity of moxidectin for the development of resistance during the decay phase are discussed.     

Effect of human tubal fluid medium and hyperactivation inducers on stallion sperm capacitation and hyperactivation

Conventional in vitro fertilization has not yet been implemented in the equine species. One of the main reasons has been the inability to develop a culture medium and incubation conditions supporting high levels of stallion sperm capacitation and hyperactivation in vitro. Although different culture media have been used for this purpose, human tubal fluid (HTF) medium, widely used in the manipulation of human and mice gametes, has not been reported so far in stallion sperm culture. The first part of this study aimed to compare HTF and Whitten's media on different stallion sperm quality and capacitation variables. Additionally, the effect of procaine, aminopyridine and caffeine in both media was evaluated on sperm motility parameters at different incubation times. Integrity and destabilization of the plasma membrane were evaluated by merocyanine 540/SYTOX Green (MC540), mitochondrial membrane potential (?Ψm) using tetramethylrhodamine methyl ester perchlorate (TMRM), acrosome membrane integrity by PNA/FITC and tyrosine phosphorylation by P‐tyrosine mouse mAb conjugated to Alexa Fluor® by flow cytometry. Motility parameters were evaluated using the integrated semen analysis system (ISAS®). We found no differences between Whitten's and HTF media and incubation time in terms of sperm viability, uninduced acrosome membrane damage or mitochondrial membrane potential at 30‐ and 120‐min incubation. Membrane fluidity (MC540) increased in both media at 30‐ and 120‐min incubation compared to noncapacitating conditions. Similarly, tyrosine phosphorylation increased in both media in capacitating conditions at 2‐ and 4‐hr incubation compared to noncapacitating conditions. Although procaine showed the best result in terms of sperm hyperactivated motility in both media, aminopyridine also showed parameters consistent with the hyperactivation including an increase in curvilinear velocity and decrease in straightness. In conclusion, HTF medium and aminopyridine equally support capacitation‐related parameters in stallion sperm.     

Antibiotic susceptibility of <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> isolated from subclinical bovine mastitis cases and in vitro efficacy of bacteriophage

Staphylococcus aureus is an opportunistic pathogen that may cause severe infections in livestock, and represents the major cause of mastitis in dairy cows. Currently, instead of using antibiotics, new strategies are sought to reduce this clinical health problem. The aim of this study was to determine the efficacy of phage therapy to kill S. aureus strains obtained from farms located at the State of Guanajuato, México. Thirty-six S. aureus strains from cow milk with subclinical mastitis were isolated and identified, and the susceptibility to antibiotics and four phages also isolated in this work was tested. It was found that more of 90% of S. aureus isolates were not susceptible to six or more antibiotics, and 100% were resistant to penicillin, dicloxacillin, cefotaxime, ampicillin and cephalothin, and 81 and 77%, to tetracycline and cefuroxime, respectively. Fortunately, 100% of S. aureus isolates were susceptible to phages used in this work, which was detected as clear zones using specific phage. It was shown for the first time, that phages used in this study are active against pathogenic S. aureus and might be incorporated into the therapy as an important tool for the control of staphylococcal bovine mastitis, specially to antibiotic-resistant S. aureus strains isolated in farm located at the state of Guanajuato, México; and its use might be extended to other regions inside or outside the country.     

Cuticular calcium penetration is directly related to the area covered by calcium within droplet spread area

Calcium penetration from salt solutions of calcium chloride and calcium acetate was measured through isolated tomato fruit (Solanum lycopersicum L., cultivar ‘Panovy’) cuticles after 4, 24, and 48 h. Droplet spread area (volume = 1 μL) and the area ultimately covered with calcium were determined with a scanning electron microscope which has an integrated energy dispersive X-ray microanalysis system. An ethoxylated rapeseed oil surfactant (RSO 5) was used for inducing different degrees of droplet spread and together with different calcium concentrations of the solutions to vary the concentration gradient in the penetration experiments. As a rule, penetration of calcium from CaCl₂ was higher than that of Ca(CH₃COO)₂. The addition of RSO 5 to salt solutions resulted in enhanced calcium penetration in comparison to unformulated solutions. However, extending penetration time from 4 to 48 h predominantly did not enhance the penetrated calcium amount significantly. Droplet spread area was enhanced up to 4-fold after adding RSO 5, whereas the area ultimately covered with calcium did not exceed twice as much. This resulted in lower percentage area covered by calcium in droplet footprints from formulated solutions in comparison to solutions without addition of RSO 5. Correlation analyses showed that penetration was not related to droplet spread area but significantly correlated with the area covered by calcium. These observations were true for unformulated calcium salts as well as for calcium solutions with addition of RSO 5. Therefore, the area ultimately covered with calcium or even other xenobiotics within a droplet footprint should be considered in developing mathematical models to simulate active ingredient (a.i.) penetration.     

Performance of dual and triple fungicide premixes for managing soybean rust across years and regions in Brazil: A meta-analysis

Soybean rust in Brazil is currently controlled with several commercial fungicide premixes composed of demethylation inhibitors (EPOXiconazole, CYPRoconazole, PROThioconazole, TEBUconazole), quinone-outside inhibitors (AZOXystrobin, TriFLoXystrobin, PYRAclostrobin, PICOxystrobin), and succinate demethylation inhibitors (BENZovindiflupyr, BIXaFen, FLUXapyroxad). Here, we summarize the performance of eight premixes evaluated in 177 cooperative trials conducted in 46 locations across 10 states from 2015 to 2020. All fungicide treatments were sprayed three times starting at R1/R2. Percentage control (, %), from back-transforming meta-analytic estimates of the log of the ratio, ranged from 56.2% (PICO + CYPR) to 76.8% (BIXF + TFLX + PROT). Estimates of mean yield difference (, kg/ha) between fungicide-treated and untreated plots were greatest for BIXF + TFLX + PROT (1,080) followed by PICO + BENZ (1,010), PYRA + EPOX + FLUX (981.5), AZOX + BENZ (910), TFLX + PROT (891), PICO +TEBU (682), TFLX + CYPR (646), and PICO + CYPR (600). Significant declines in both and in as little as 4 years were detected for AZOX + BENZ (35.3%; 550 kg/ha) and PICO + BENZ (15.5%; 359.8 kg/ha). Variance in was reduced by the inclusion of baseline severity as covariate. In trials where baseline disease was ≥70%, yield was 250 kg/ha greater compared to areas with low baseline disease. Disease control and yield response were generally greater in the south-east, where the frequency of profitable scenarios was 30% higher on average than in the north-west. Results of this meta-analysis are critical for supporting decisions during planning of fungicide programmes.     

Links between riparian landcover,instream environment and fish assemblages in headwater streams of south‐eastern Brazil

We hypothesised and tested a hierarchical organisation model where riparian landcover would influence bank composition and light availability, which in turn would influence instream environments and control fish assemblages. The study was conducted during the dry season in 11 headwater tributaries of the Sorocaba River in the upper Paraná River Basin, south‐eastern Brazil. We focused on seven environmental factors each represented by one or multiple environmental variables and seven fish functional traits each represented by two or more classes. Multivariate direct gradient analyses suggested that riparian zone landcover can be considered a higher level causal factor in a network of relations that control instream characteristics and fish assemblages. Our results provide a framework for a hierarchical conceptual model that identifies singular and collective influences of variables from different scales on each other and ultimately on different aspects related to stream fish functional composition. This conceptual model is focused on the relationships between riparian landcover and instream variables as causal factors on the organisation of stream fish assemblages. Our results can also be viewed as a model for headwater stream management in that landcover can be manipulated to influence factors such as bank composition, substrates and water quality, whereas fish assemblage composition can be used as indicators to monitor the success of such efforts.