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The aim of the present study was to evaluate semen cryopreservation with ACP-Lact® diluent, which consists of coconut water powder (ACP) added to goat milk powder. After thawing, the samples were evaluated for sperm kinetics, membrane evaluation and in vivo insemination. For cryopreservation, a pool was made with the ejaculate of six goats, diluted in four equal aliquots for the respective treatments: T1 (ACP-Lact®); T2 (ACP-Lact® 50%); T3 (ACP + 2.5% egg yolk) and T4 (Tris + 2.5% egg yolk). After dilution of the treatments, the samples were placed in 0.5 ml straws and chilled at a rate of -1.07°C/min. After reaching 4°C and stabilizing for one hour, the straws were placed in nitrogen vapour at -60°C for 15 minutes and then immersed in liquid nitrogen (-196ºC). The straws were thawed in a 37°C water bath and kinetic assessments were performed immediately using a computerized semen analysis program (CSA), viability (EN), membrane functionality (HOST), mitochondrial activity (DAB) and DNA integrity assessment of spermatozoa. For the in vivo experiment, ten goats were inseminated, divided into two groups of five goats each, G1 inseminated with ACP-Lact® and G2 with ACP, by fixed-time artificial insemination (FTAI). Regarding the kinetic parameters, the ACP-Lact® treatment showed higher progressive motility (PM) and sperm velocity than the other treatments (36.77%). In the VSL parameter the ACP-Lact diluent was superior to ACP and Tris. In viability the treatment with ACP-Lact® was superior to the treatment with Tris, 95% and 83% respectively. In FTAI two goats were born out of the 5 goats inseminated with ACP-Lact®. It was concluded that the use of ACP-Lact® for cryopreservation of caprine semen is efficient in maintaining seminal parameters during thawing in vitro and in vivo and proved to be a good alternative extender for the caprine species., Competing Interests: Conflicts of interest: The authors have no conflict of interest to declare.

This study investigated platelet-rich plasma (PRP) and adipose stem cell-conditioned medium (ASC-CM) use as a strategy to accelerate tissue healing. Platelet-derived growth factor (PDGF) and vascular endothelial growth factor (VEGF) were quantified in fresh and freeze-dried PRP and ASC-CM, and a stability test was performed in the freeze-dried samples (90 and 180 days of storage). A cell proliferation test was performed using equine mesenchymal stem cell culture in reconstituted PRP gel mesh after freeze-drying. In vivo PRP, ASC-CM applications, or their association were performed in induced wounds at 15 and 9-day intervals, according to the treatments: saline solution (control), PRP, ASC-CM, or ASC-CM + PRP. Horses were monitored through photographs and wound area measurements on days 5, 7, 15, and 24 after lesion induction. Skin biopsies were obtained on days 15 and 24 of the experiment. PDGF and VEGF quantification did not differ between fresh or freeze-dried treatments, was similar after freeze-drying or 90 days of storage, but showed a significant reduction after 180 days of storage. Comparing all treatments, no differences were observed in the histopathological analyses. For inflammation, fibroplasia, and collagen formation, only the time effect between the first and second biopsies was significant. The cell proliferation test revealed intense multiplication in the PRP gel mesh. Healing time was similar among all treatments. In conclusion, our results showed the possibility to produce and maintain freeze-dried PRP and ASC-CM for 90 days. Further studies are needed to better explore the in vivo therapeutic PRP and ASC-CM effects., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Effects were assessed of the dilutants TRIS and ACP - 101c® with the addition of different guinea fowl ( Numida meleagris ) egg yolk concentrations. Fifteen ejaculates were collected from five goats of the Anglo Nubian breed. The ejaculates were pooled and then divided into 12 groups, two control groups (GC1 TRIS, with 2.5% Gallus gallus domesticus hen egg yolk GOGD), (GC2 Control Group ACP - 101c®, with the addition of 2.5% Gallus gallus domesticus hen egg yolk GOGD) and ten experimental groups (EG), containing TRIS and ACP added with different concentrations of egg yolk from guinea hen ( Numida meleagris ) (TRIS 2,5% GONM; TRIS 5% GONM; TRIS 10% GONM; TRIS 15% GONM; TRIS 20% GONM; ACP® 2,5% GONM; ACP® 5% GONM; ACP® 10% GONM; ACP® 15% GONM; ACP® 20% GONM). Then cryopreservation was carried out and the samples stored in liquid nitrogen (-196 °C). After seven days, the samples were thawed and assessed for spermatic kinetics, immunofluorescence and sperm morphology. Analysis of GOMN by the CASA system showed that the various parameters were similar to those of GOGD (P>0.05). The membrane integrity, mitochondrial potential and the acrosome were not influenced by the treatment (P>0.05) nor by the dilutant used for cryopreservation (P>0.05). The spermatic morphology was also preserved by the different GOGD and GONM concentrations in the ACP® and TRIS dilutants, with no statistically significant differences (P<0.05). It was concluded that Numida meleagris egg yolk, as external membrane cryoproctant added to the dilutants ACP-101c® and TRIS, improved goat semen quality., Competing Interests: Conflicts of interest: The authors have no conflict of interest to declare.

The study aimed to evaluate the effect of powdered coconut water-based diluent (ACP-101c) associated with extra virgin coconut oil (CO) as an external cryoprotectant in the conservation of cryopreserved buck sperm. For cryopreservation, the ejaculates from four bucks were pooled and divided into three aliquots and diluted at 37°C for treatments T1 (ACP-101c + 2.5% egg yolk +7% glycerol), T2 (ACP-101c + 2.5% CO +7% glycerol), and T3 (ACP-101c + 5% CO +7% glycerol). Then, the samples were packaged and cooled at a rate of 1.07°C/min decrease. Upon reaching 4°C, the samples were stored in a refrigerator at 4°C for 30 minutes for stabilization. After this period, the straws were frozen in nitrogen vapor for 15 minutes and then immersed and stored in liquid nitrogen at -196°C. After thawing, the samples were evaluated for sperm kinetics, plasma membrane integrity, acrosomal integrity, membrane functionality, mitochondrial activity (MA), and sperm morphology. In this study, no statistically significant differences were observed between the three treatments regarding the kinetic parameters ( p  > 0.05; Table 1). However, in relation to the velocities, a reduction was observed beyond the expected. There were no statistically significant differences between the diluents T1, T2, and T3 for the three velocities (curvilinear velocity [VCL], linear velocity [VSL], average path velocity [VAP]). Furthermore, no statistically significant differences were observed ( p  > 0.05) among treatments regarding the evaluation of membrane integrity, the functional membrane, MA, and sperm morphology after thawing. In conclusion, the use of CO in concentrations of 2.5% and 5.0% is effective in maintaining goat sperm quality, presenting itself as an alternative diluent for international programs of artificial insemination and embryo transfers. [Table: see text].

Purpose: The aim of this study was to evaluate the physical and chemical characteristics of coconut water and to analyze the use of coconut water solution for the conservation of human corneas., Methods: This was an experimental and controlled study performed at the Eye Bank of the General Hospital of Fortaleza. The coconut water-based solution was prepared at the Goat Seed Technology Laboratory of the Department of Veterinary Medicine of the State University of Ceará. Discarded corneas from the Eye Bank were divided into two groups for sequential experiments: G1, coconut water-based solution (experimental group), and G2, conservative treatment with OPTISOL GS® (control group). The osmolality of corneas in G1 was analyzed sequentially at 275, 300, 325, 345, 365, and 400 mOsm/L. The viability of the corneas was determined by specular microscopy and biomicroscopy on the first, third, and seventh days., Results: Corneas preserved in a solution of 365 and 345 mOsm/L had a transparency of 8 mm until the third day and had diffuse edema in the periphery, central folds, and partial epithelium loss until the seventh day. The 365-mOsm/L solution was associated with the worst results during follow-up. Corneas placed in Optisol-GS retained their original aspects., Conclusions: Coconut water-based preservative partially maintained corneal transparency and epithelial integrity, especially during the first three days of follow-up. The coconut water-based solutions used were not effective for use as preservatives in a human eye bank.

The objective of this study was to test the efficiency of powdered coconut water (ACP-406 ® ) base-medium without or with the addition of supplements on in vitro culture of isolated goat secondary follicles. Follicles were cultured for 18 days in α-MEM or in ACP-406 ® , both without supplements (referred to as α-MEM and ACP, respectively), or both supplemented with BSA, insulin, transferrin, selenium, glutamine, hypoxanthine, and ascorbic acid (referred to as α-MEM + and ACP + ). Follicular morphology, antrum formation, follicular and oocyte diameter, levels of glutathione (GSH), and chromatin configuration after in vitro maturation were evaluated. At the end of culture, ACP-406 ® base-medium (without or with supplements) showed a higher (P < 0.05) percentage of normal follicles than α-MEM (without or with supplements). Antrum formation was similar among α-MEM + , ACP and ACP + , and significantly higher than α-MEM without supplements. The follicular diameter was greater in ACP + than α-MEM, and similar to other treatments. Moreover, fully and daily grown rates were higher (P < 0.05) in ACP-406 ® base-medium (without or with supplements) than α-MEM (without or with supplements). Levels of GSH were similar between ACP + and α-MEM + treatments. Both ACP + and α-MEM + allowed meiotic resumption without a significant difference between the two groups. In conclusion, supplemented ACP-406 ® base-medium maintained follicular survival and promoted the development as well as meiotic resumption of isolated goat secondary follicles cultured in vitro for 18 days., Competing Interests: Conflicts of interest: The authors have no conflict of interest to declare., (Copyright © The Author(s).)

Background/aim: A newly developed powdered coconut water formulae (ACP-404) has been proposed to preserve periodontal ligament cell viability. The aim of this study was to investigate the periodontal healing of dog teeth replanted after storage in ACP-404 with 2 different osmolalities., Materials and Methods: Sixty teeth of 10 adult beagle dogs were extracted and randomly subjected to the following protocols: immediate replantation after extraction (positive control); dry conditions (negative control); and storage in ACP-404 I (G1), ACP-404 II (G2), in coconut water long shelf life (G3) or in pH-neutralized coconut water long shelf life (G4) for 50 minutes before replantation. The animals were euthanized 28 days post-operatively and processed for histomorphometric analysis using an optical microscope. The groups were subjected to histomorphometric analysis compared with immediate replantation and dry conditions. The data were statistically analysed using the Kruskal-Wallis test with post-analysis using Dunnett's test., Results: The normal periodontal area was significantly higher for the G1 than for the G2 and G3 groups (P < .001). The positive control group presented similar normal periodontal areas compared to the experimental groups, except for the G3 and negative control groups (P < .001). G1 presented the lowest level of external root replacement resorption compared to the other groups (P < .05). All storage solutions had similar periodontal conditions for repaired resorption areas, inflammatory root resorption, ankylosis and replacement resorption (P > .05)., Conclusions: ACP-404 I (G1) was better overall and was able to preserve the normal periodontium and had less external root replacement resorption., (© 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

The use of frozen semen for artificial insemination is the main approach utilised for the genetic improvement of most domesticated species. The advantages include lower transportation costs, continuous availability of semen, fewer occurrences of sexually transmitted diseases and the incorporation of desirable genes in a relatively short amount of time. Nevertheless, the use of frozen semen in buffalo herds remains limited due to the loss of sperm quality when buffalo semen is frozen. So, the goal of this study was to evaluate the pre- and post-cryopreservation quality of buffalo semen diluted in three distinct freezing media: Tris-egg yolk, Botu-bov® (BB) and ACP-111®. Thirty-two ejaculates from four bulls were analysed in terms of kinetics, morphology and sperm viability by epifluorescence microscope. Thawed samples were also evaluated for capacitation-like damage, DNA fragmentation and plasma and acrosomal membrane integrity using flow cytometry. The Tris-egg yolk and BB® extenders yielded better results than the ACP-111® extender for kinetics parameter (total motility, progressive motility and percentage of rapid cells). However, semen samples were similar for parameters evaluated by flow cytometry. Taken together, the data indicate that in comparison with Tris-egg yolk and BB extender, ACP-111® can also be used as an extender for buffalo semen cryopreservation., (© 2017 Blackwell Verlag GmbH.)