BACKGROUND: Storing semen at a temperature of 4 degrees Celsius reduces its motility and quality.OBJECTIVES: This study aimed to determine the effect of adding calcium compounds, including calcimaphor (CMP), calcium gluconate (CG), and calcium chloride (CaCl2), on the motility and progressive motility of rooster sperm kept at refrigerator temperature.METHODS: This research used five pieces of 45-week-old Lari breed roosters. The liquid diluent added different calcium compounds at 0.56, 0.056, and 0.0056 mM concentrations. After treatment, the diluted seminal samples were cooled at the storage temperature to avoid thermal doubt and then transferred to the refrigerator at 4 degrees Celsius. Parametric factors that were more important such as the percentage of laterality and progressive mobility, were measured visually using the lens of a 40-light microscope, survival was checked using the eosin-nigrosin staining method, acrosome health percentage by formalin citrate method, cell membrane health with hypoosmotic test, lipid peroxidation level, fertility was evaluated using perivitelline membrane sperm reaction 72 hours after storing at 4 degrees Celsius.RESULTS: Based on the results, different calcium compounds in most concentrations could significantly affect the parameters of survival, mobility, and progressive mobility (P<0.05).CONCLUSIONS: Most of the sperm-related parameters in the control group decreased after this period of storage in the refrigerator, but the addition of calcium gluconate (0.56, 0.056, and 0.0056), Calcimaphor (0.56, 0.056, and 0.0056) and calcium chloride (0.56, 0.056 and 0.0056) to the semen thinner maintained the quality indicators of rooster sperm.

Three ejaculates from each of 3 stallions were used to evaluate a computerized system (Hamilton-Thorn motility analyzer; HTMA) for measuring equine spermatozoal motility. Variance components (ejaculate-within-stallion, chamber-within-ejaculate, and microscopic field-within-chamber) were determined for each stallion after diluting ejaculates to 25 X 10(6) spermatozoa/ml with a skim milk-glucose seminal extender. The HTMA was compared with frame-by-frame playback videomicrography (VIDEO) for determining: percentage of spermatozoal motility and spermatozoal number in microscopic fields; curvilinear velocity and straight-line velocity of individual spermatozoa for 5 track types; and repeatability of those velocity measurements. The effect of spermatozoal number per microscopic field on incidence of intersecting spermatozoa and the outcome of intersecting spermatozoa also were evaluated. Greatest variability in motility measures was generally attributed to the microscopic field-within-chamber component. The HTMA was highly correlated with VIDEO for estimation of spermatozoal numbers per microscopic field (r = 0.99; P <0.001) and motility (r = 0.97; P <0.001); however over the entire range of spermatozoal numbers, the HTMA yielded higher spermatozoal numbers per microscopic field (P <0.05) and higher motility (P <0.05) than did VIDEO. The HTMA- and VIDEO-derived measurements of curvilinear and straight-line velocities were highly correlated for all spermatozoal track types, but both measures were higher (P <0.05) by use of the HTMA than by use of VIDEO for most track types. For 3 of 5 track types, measurements of curvilinear and straight-line velocities were less variable (P <0.05), using the HTMA, rather than VIDEO. Using the HTMA, the number of intersecting spermatozoa was highly correlated with spermatozoal numbers per microscopic field (r = 0.97; P <0.001). The percentage of erroneous track interpretations involving intersecting spermatozoa was high (85.3 +/- 2.7%). The HTMA was a reliable system for determining percentage of spermatozoal motility and velocity measures in video recordings of equine semen diluted to spermatozoal concentration of 25 X 10(6)/ml prior to evaluation.

Properties of the attachment of Treponema hyodysenteriae to Henle intestinal epithelial (HIE 407) cells were examined. The frequency of attachment depended on the motility and viability of the spirochetes. Rabbit hyperimmune and swine convalescent antisera inhibited attachment. Treatment of HIE cells with neuraminidase had no effect on attachment; however, treatment of spirochetes with the enzyme decreased adherence significantly (P = 0.01). Attachment was inhibited by N-acetylneuraminic acid, D-glucuronic acid, and fetuin. Adherence was increased following coincubation with N-acetylglucosamine or yeast mannan. Surface antigens of T hyodysenteriae, isolated by chemical extraction, competitively inhibited adherence. Concentrated T hyodysenteriae culture supernatant fractions inhibited adherence, but concentrated phosphate buffered-saline washings of the spirochete and concentrated uninoculated media did not inhibit adherence. Sialic acid was detected in unwashed T hyodysenteriae and spent culture supernatant fractions in higher concentrations than from washed spirochetes and uninoculated media. It was concluded that the binding adhesins on T hyodysenteriae for cultured HIE cells may contain sialic acid residues.

Two hundred seventy-eight strains of Salmonella typhimurium isolated from 1973 to 1981 from animal sources in New York State were studied for possible virulence determinants and for a serotype-specific plasmid possibly linked with virulence. Of the strains, 98% possessed type-1 fimbriae. All strains possessed flagella and were motile. One hundred twenty-three strains (44%) treated with mitomycin C tested positive for the cholera-Escherichia coli heat labile family of toxins by a kinetics-based ELISA; when treated with mitomycin C and extracted with polymyxin B, 249 (90%) were positive in the kinetics-based ELISA. All strains were negative in the Biken Test. A smooth cell wall was found in 99% of the strains. Sixty-one percent (169) of the strains had a 62-Md plasmid. Seventy-six (27$%) of the strains had detectable plasmids ranging in size from 1 to 124 Md.

Objective: To improve the fertilizing ability of frozen buffalo semen using the more beneficial, accurate, and cheap technique of laser irradiation at specific wavelengths and exposure times. Materials and Methods: The red laser source (625 nm) was used in this study with 5 watts output power and for the irradiation of the semen samples for 5 min; the laser focus spot area was 1 cm2. Thirty straws belonging to five buffalo bulls were used in this study. Results: The results show that total motility (%) and progressive motility (%) increased insignifi¬cantly after 5 min of exposure (73.8 ± 1.4 and 60.4 ± 1.1, respectively) compared to the control sample (70.9 ± 0.9 and 57.5 ± 1.7, respectively). All velocity parameters (velocity average path, ve¬locity curved line, and velocity straight line μm/sec) recorded a significant (p &lt; 0.05) increase in samples measured 5 min after exposure (52.3 ± 1.3, 83.5 ± 2.0, and 43.5 ± 1.2, respectively) com¬pared to the untreated ones (47.1 ± 2.0, 76.3 ± 3.1, and 38.6 ± 1.9, respectively). Conclusion: The application of the red laser light on buffalo semen postthawing resulted in a positive correlation with almost every motility parameter; it may be recommended to apply this technique pre-in vitro fertilization for embryo production of buffalo species. [J Adv Vet Anim Res 2022; 9(3.000): 396-404]

Objective: This study aimed to measure the anthelmintic effects of betel nut (Areca catechu) and neem (Azadirachta indica) leaf extracts against Fasciola spp. in vitro in comparison with the com-mercial dewormer, Albendazole, and the negative control, nutrient broth. The study determined the extract concentration that produced the highest efficacy based on the average recorded mean motility time, gross, and microscopic changes of the flukes treated with different concentrations of plant extracts. Material and Methods: The study consisted of eight treatments. Every treatment consisted of 10%, 20%, and 40% concentrations of both betel nut extract (BNE) and neem leaf extracts, positive control treatment (Albendazole-treated) and negative control treatment (25 ml nutrient broth). The motility of the flukes on all treatments was based on the established motility criteria scoring. The flukes subjected to all treatments were processed for histopathological analysis. Results: The result of the study revealed that after exposure of Fasciola spp. under 10%, 20%, and 40% extract concentrations, betel nut showed higher efficacy having the recorded mean motility time of 0.22, 0.07 min, and no movement upon contact, respectively, than Albendazole which produced mean motility time of 0.38 min. Nevertheless, the flukes treated with 10%, 20%, and 40% neem leaf extracts obtained the average mean motility time of 220, 151, and 98 min, respectively. Conclusions: The results gathered showed that 40% BNE concentration showed the highest efficacy based on the recorded mean motility time. All treatments of betel nut extract evidently showed marked changes in the gross and microscopic morphology of the flukes. However, the neem extract was ineffective in all concentrations although changes were observed microscopically. Furthermore, the nutrient broth was proven to be effective as a culture medium since the flukes remained active until 8 h of exposure. [J Adv Vet Anim Res 2019; 6(1.000): 44-49]

Objectives: This study aims to determine the effect of a toll-like receptor 7/8 (TLR7/8) ligand on the motility of putative X- and Y-chromosome-bearing sperm in rams. Materials and Methods: Sperm from three fertile rams were incubated with tris-citrate buffer containing 0 to 0.9 μM resiquimod (a TLR7/8 ligand) that affects only the X chromosome sperm. Sperm was then subjected to a swim-up test method. After incubation, sperm in the top and lower layers of the media were transferred into a separate new tube. Sperm motility characteristics, concentration, morphometry, and adenosine triphosphate (ATP) concentration from both layers were identified. The data were analyzed for multiple comparisons using one-way analysis of variance. Results: The study demonstrated that the presence of resiquimod used in the swim-up method facilitated the morphometric separation of smaller sperm, predominantly representing Y chromosome sperm in the top layer. Meanwhile, larger samples, indicating X chromosome sperm, accumulated in the lower layer of the media, thus increasing sperm concentration. Resiquimod also decreased motility in the lower layer, but samples in the top layer were unaffected. This decrease was reinforced by the depletion of the ATP level of sperm at the lower level. Conclusion: The TLR7/8 ligand reduced the motility of the putative X-sperm by decreasing their ATP content, allowing separation from the putative Y-sperm. These results suggested the importance of TLR7/8 as a potential biomarker in sperm selection technology. [J Adv Vet Anim Res 2024; 11(3.000): 648-654]

Objective: To explore the effect of glycerol at different concentrations using different extenders on DNA fragmentation and motility of frozen-thawed Kintamani Bali dog spermatozoa. Materials and Methods: Sample was collected from four mature Kintamani Bali dogs. Each ejacu¬late was prepared for cryopreservation with two different semen extenders; egg yolk Tris extender and coconut water-based extender. For each extender, three different glycerol concentrations were used; 4%, 6%, and 8%. Each of the six aliquots was loaded into 0.5 ml cryotube, placed on a styrofoam box 5 cm over liquid nitrogen for 10 min, and immersed in liquid nitrogen up to 8 min. Then, the frozen cryotubes were transferred into liquid nitrogen container. The cryotubes were thawed in a water bath at 38.5&#xb0;C for 120 sec. After equilibration and thawing, each sample was assessed for motility parameters and for DNA fragmentation. Results: The addition of 6% glycerol to extenders revealed the most effective addition of glycerol on motility and sperm DNA fragmentation after equilibrium and post-thawing. Conclusion: It is concluded that both extenders with the addition of 6% glycerol are safe to be used as an extender in Kintamani Bali dog semen preservation, and DNA fragmentation of Kintamani Bali dog spermatozoa was not influenced by the freezing procedure. [J Adv Vet Anim Res 2019; 6(2.000): 158-162]

Objective: The objective of this study was to evaluate the effects of caffeine and taurine on the motility and viability of chilled equine semen. Materials and Methods: A total of 12 ejaculates were collected from three mature stallions with proven fertility during the breeding season. The gel-free spermatic fraction of each ejaculate was divided into two aliquots and diluted with a semen extender (either INRA 96® or BotuSemen Gold®). The aliquots were then split and assigned to one of the six treatment groups: control (no supplement), caffeine (2 and 4 mM), taurine (25 and 50 mM), and a combination of caffeine (2 mM) plus taurine (25 mM). Samples were stored at 4&#xb0;C and analyzed at different time points (0, 24, 48, 72, and 96 h) to evaluate total (TMOT) and progressive (PMOT) motility and viability by computer-assisted sperm analysis. Results: Regardless of the extender, PMOT and TMOT decreased over time. However, compared with the control, the treatment with 4 mM caffeine significantly mitigated the decrease in PMOT at 72 h. Additionally, semen treated with a combination of caffeine plus taurine maintained a significantly higher PMOT at 96 h, with improved viability at all time points. Conclusions: The combination of caffeine plus taurine helps maintain chilled equine semen viabil¬ity and progressive motility up to 96 h independently of the extender used. [J Adv Vet Anim Res 2021; 8(4.000): 635-641]