Characterization of Nucleated Cells From Equine Adipose Tissue and Bone Marrow Aspirate Processed for Point-of-Care Use

The objective of this study was to compare nucleated cell fractions and mesenchymal stromal cells (MSCs) from adipose tissue to bone marrow processed by a point-of-care device that are available for immediate implantation. A paired comparison using adipose and bone marrow from five horses was done. The number of nucleated cells, viability, total adherent cells on day 6 of culture and colony-forming unit fibroblasts (CFU-Fs) were determined. Gene expression for markers of stemness, adipogenic, chondrogenic, osteogenic lineage, and collagen formation was measured in total RNA isolated from adherent adipose and bone marrow cells. Day 6 adherent adipose-derived MSC was frozen briefly, whereas day 6 adherent bone marrow–derived MSC was passaged two additional times to obtain adequate cell numbers for chondrogenic, osteogenic, and adipogenic cell differentiation assays. The total cell count per gram was significantly greater for bone marrow, whereas total adherent cells per gram and the CFU-F per million nucleated cells on day 6 were significantly greater for the adipose. In undifferentiated adherent cells, relative gene expression for CD34, adipogenic, and chondrogenic markers and collagen II was significantly lower in the adipose-derived cells. Conversely, expression of collagen I was significantly higher in the undifferentiated adipose-derived cells. Cell density and total RNA were higher in differentiated adipogenic and osteogenic cultures of adipose cells and in chondrogenic cultures of bone marrow cells. This cell preparation method provides a stromal vascular fraction with a large proportion of multipotent MSCs. There are differences in the cells obtained from the two sources. This method can provide an adequate number of multipotent cells from adipose tissue for immediate implantation.     

Studies on the use of hydrogen peroxide as a method for the control of sea lice on Atlantic salmon

Atlantic salmon (Salmo salar) post-smolts exposed to 1.23 hydrogen peroxide for 20 min at 13.5 C suffered an acute toxicity resulting in a 35% mortality within 2 h. Under similar conditions at 10 C no mortalities were observed with Atlantic salmon or goldsinny wrasse (Ctenolabrus rupestris). No histological changes were noted in tissues from exposed fish. Thirty-three per cent of adult and pre-adult sea lice (Lepeophtheirus salmonis) were immobilized or killed following exposure to 0.5 hydrogen peroxide at 10 C, rising to 98% at 2. Some lice were able to recover and regained normal swimming movements. Gas bubbles within the haemolymph caused affected lice to float on the water surface. A delay in the toxicity of hydrogen peroxide to copepodites occurred, with a 10% mortality following a 20 min exposure to 1.25 at 10 C rising to 100% mortality at 19 h post treatment.Dilute hydrogen peroxide was stable over the 20 min treatment period. Aeration and higher temperatures increased the long-term breakdown of a working concentration of hydrogen peroxide in seawater.     

Incidental capture and mortality of sea turtles in the industrial double‐rig‐bottom trawl fishery in south‐eastern Brazil

1. Incidental capture by fisheries is one of the principal threats to sea turtles. This study analysed spatial and temporal patterns of sea turtle bycatch, and estimated the direct initial mortality rate of these animals, in the industrial double‐rig‐bottom trawl fishery in south‐eastern Brazil. This is also the first attempt to relate bycatch/at‐sea mortality in bottom trawling to stranded turtles found along the adjacent coast.
2. The fishery was monitored from October 2015 to April 2018 through data collected voluntarily by the captains of eight industrial double‐rig trawlers. Two hundred and one sea turtles were captured during 9362 tows (43,657.52 trawling hours), resulting in a catch per unit effort (CPUE) of 0.0025 ± 0.0032 turtles h^?1 with a standard net of 30.5 m headrope, with no significant difference between the estimated CPUEs for licensed shrimp and demersal fish trawlers.
3. Caretta caretta (52.24%) and Lepidochelys olivacea (38.81%) were the most frequently captured species. According to Generalized Linear Models, C. caretta bycatch was significantly higher during winter, at lower latitudes (?24° to ?23°) and higher longitudes (?42° to ?40°), while the L. olivacea bycatch was significantly higher at higher latitudes (?23° to ?21°). The direct initial mortality rate of sea turtles in the shrimp trawlers was 7.65 ± 3.85%. However, none of the dead individuals subsequently released with plastic tags (n = 10) were found stranded on the coast. Mortality was not significantly related to the depth or duration of the trawling.
4. The results of this study suggest the need for improvements to the current management of the bottom trawl fishery in Brazil, moving from a species‐based to a spatial and seasonal‐based approach. There is also a need to develop turtle excluder devices adapted to local fishing conditions.

     

A simple and easy method to measure ammonia volatilization: Accuracy under field conditions

Field studies on soil ammonia(NH₃)volatilization are restricted in many countries owing to the high costs commonly demanded for accurate quantification.We assessed the accuracy of a simple,open chamber design to capture NH₃under field conditions,as affected by different chamber placement schemes.Urea-15 N was surface applied to lysimeters installed in the spaces between maize rows.Open chambers made from plastic bottles were installed on each lysimeter with variations in i)N rates(3,8,13,and 18 g m^-2),ii)the height of the chamber above the soil surface(0,5,and 10 mm),and iii)chamber relocation(static vs.dynamic).Reference lysimeters without chambers were used to measure NH₃losses by¹⁵N-balance.Losses of NH₃-N accounted for more than 50%of the applied N.Relocation of the chambers had no impact on their NH₃-trapping efficiencies,proving to be an unnecessary procedure.Variation in the height of the chambers above the soil surface affected the capture of NH₃,but the results still maintained high linearity with the NH₃losses quantified by the reference method(R²>0.98).When the same placement scheme used in the introductory study describing the chamber was utilized(static and touching the soil surface),we found a trapping efficiency of 60%,which was very similar to that(57%)obtained in the previous study.Our results show that this simple,open chamber design can be used with satisfactory accuracy under field conditions,provided that simple,standardized procedures are warranted.     

Fitness characteristics of the European lineages of Phytophthora ramorum

As an introduced pathogen, Phytophthora ramorum exists as four near-clonal evolutionary lineages, of which only EU1 and EU2 are established in the UK. EU1 has become widespread since the first findings in 2002 whereas EU2, detected in 2011, has a more limited distribution. Both lineages are epidemic in plantation-grown larch, sporulating asexually on needles, but also causing heavy dieback and mortality. To understand whether EU1 and EU2 pose different threats to forest health, we compared their growth characteristics on agar, pathogenicity on several hosts, and sporulation on Japanese larch needles. When pathogenicity was evaluated by measuring colonization at 20 °C in mature bark (phloem) of Japanese and European larch (Larix kaempferi and L. decidua), English oak (Quercus robur), and beech (Fagus sylvatica), Japanese larch was the most susceptible and oak the least susceptible. On average, EU2 isolates produced significantly larger lesions than EU1 isolates in Japanese larch and oak although not in the other hosts. With tests using young saplings of Japanese and European larch, damaging bark lesions formed at both 10 °C and 20 °C, but EU2 was significantly more pathogenic at 20 °C on both hosts compared with EU1. In contrast, both lineages caused similar amounts of necrosis on inoculated leaves of rhododendron (Rhododendron ponticum). Moreover, EU2 isolates usually sporulated less abundantly on larch needles compared with EU1 isolates, suggesting a trade-off in pathogenicity and sporulation between lineages. As EU2 tends to have smaller sporangia than EU1, this could also reduce the inoculum potential of EU2.     

Ovarian evaluation of Girolando (Holstein × Gir) heifers submitted to a GnRH–PGF2α–GnRH protocol in the dry or rainy seasons in the tropical savannah

The Girolando breed is used in pasture-based dairy production systems in Brazil to associate the high production of Bos taurus to the rusticity and thermal adaptation of Bos indicus. This study was designed to evaluate the physiological response to a gonadotropin-releasing hormone (GnRH)–prostaglandin F_2α (PGF_2α)–GnRH protocol to synchronize the ovulation in 40 Girolando heifers of a pasture-based dairy production system and its relationships with the temperature and humidity index (THI) during the dry (DS) and rainy season (RS) in the tropical savannah—Brazil's cerrado biome. Responses were characterized by follicular and corpus luteum number and diameter, ovulation (D9), and pregnancy rates after first AI. Total follicle number (8.1?±?0.3?×?8.8?±?0.3), D9 ovulatory follicle diameter (11.9?±?0.4?×?10.1?±?0.4 mm), corpus luteum diameter (8.6?±?1.3?×?3.9?±?1.5 mm), corpus luteum score (3.7?±?0.8?×?1.8?±?1.0), corpus luteum diameter after AI (9.6?±?1.6?×?3.9?±?1.5 mm), and corpus luteum score after AI (3.2?±?0.4?×?0.9?±?0.6) in DS and RS differed (P?<?0.01). D9 ovulation rate was 40 % (DS) and 20 % (RS), without differences (P?>?0.05). Pregnancy rate was 45 % (DS) and 11 % (RS), with differences (P?<?0.01). THI differed between DS and RS (P?<?0.01). THI may interfere in the follicular and luteal dynamics and in the response of Girolando heifers to the GnRH–PGF_2α protocol in the tropical savannah, thus reducing the chances of pregnancy at the first artificial insemination.     

A Case of Atypical Psoroptic Mange in a Domestic Rabbit

A female checkered giant cross rabbit (Oryctolagus cuniculus) was presented with skin lesions on the perineal region. The cutaneous lesions extended 2 cm from the vulva and perineal glands. They were moist and erythematous, with abundant greasy yellowish scabs. Skin scrapings from the lesions were mounted in Amman’s chloral-lactophenol on microscope slides and examined at ×10 and ×40 magnification. Numerous mites were observed and identified as Psoroptes cuniculi, a common causal agent of otitis externa in rabbits. The rabbit was treated successfully with ivermectin. This case of psoroptic mange is atypical owing to the perineal location, apparent absence of auricular lesions, and the lack of contagion despite close contact with several other rabbits.