Biometrical and histomorphometrical changes of testis in the dynamics of postnatal ontogenesis from birth to puberty of Black Bengal goat.

Objectives: The study aimed to account for baseline biometrical and histomorphometric testicular changes in Black Bengal goats during postnatal development. Materials and Methods: Black Bengal goats, divided into group I of VII; day 0; 1, 2 weeks; 1, 2, 4, and 6 months of age, respectively, were used in this study. Results: The biometrical and histomorphometric values of the testis varied significantly (p < 0.05) from postnatal 1-2 months. From day 0 to 2 months, seminiferous tubules, called sex cords, contained simply peripherally placed Sertoli cells and centrally placed gonocytes. Gonocytes, positioned in the center, moved centrifugally in the direction of the basement membrane of sex cords with the advancement of age, transformed into prespermatogonia, and were distributed among the Sertoli cells at the edge of sex cords that make up the basal cell layer in nearly all of the seminiferous tubules by 2 months after birth. Initiation of spermatogenesis, i.e., stratification and lumination of seminiferous epithelium, took place in the 4th months. At 6 months, all types of spermatogenic cells had been identified. The onset of puberty, i.e., the establishment of spermatogenesis, was noticed to have been established at 6 months of postnatal age in Black Bengal goats, as shown by the spermatozoa that were adhered to the ad luminal border of the Sertoli cells and also in the tubular lumen. Conclusion: This research is the first to document the varying biometrical and histomorphometric measurements of the testis in Black Bengal goats from birth to puberty.

Effects of glucose and trehalose on tris-citric acid-egg yolk-fructose diluents for semen cryopreservation in goat.

Objectives: This study aimed to examine the impacts of the wide range of concentrations of glucose and trehalose on the tris-citric acid-egg yolk-fructose (TCEF) extenders for cryopreservation of goat semen. Materials and Methods: The sperm sample was pooled, washed, and diluted in control (TCEF without glucose and trehalose), TCEF + glucose (75, 150, 450, and 900 mm), and TCEF + trehalose (75, 150, 450, and 900 mm). After equilibrations, the semen straws were frozen under LN2 in the LN2 tank. After LN2 storage, the straws were thawed at 37°C for 30 seconds. The sperm parameters of all study groups were checked after equilibration and freezing. Results: After equilibration, the progressive motility (PM), total motility (TM), and viability of sperm in G-75, G-150, G-450, T-75, T-150, and T-450 were not significantly different (p < 0.05) from those in control. After cryopreservation and thawing, the PM, TM, and plasma membrane integrity (PMI) of T-150 were significantly higher (p < 0.05) than in control, G-75, G-900, T-75, and T-900. The viability of sperm in T-150 was substantially higher (p < 0.05) than in the control, whereas there was no significant difference among the control, G-75, G-900, T-75, and T-900. However, the acrosome integrity (AI) of sperm in G-900 was significantly decreased (p < 0.05) compared to the control, G-75, G-150, G-450, T-75, T-150, and T-450. Conclusion: According to the findings, the supplementation of 150 mm trehalose in the TCEF diluent was more efficient for sperm cryopreservation in the buck as reflected by PM, TM, viability, PMI, and AI.

Review of anthrax: A disease of farm animals.

Anthrax is a rapidly fatal infectious disease affecting herbivores and people. In the farm animals, cattle and sheep are more susceptible, followed by goats and horses, while dwarf pigs and Algerian sheep are relatively resistant. Bacillus anthracis, the causative agent of anthrax, produces spores and persists for decades in the soil, initiating an outbreak through a favorable climate shift. Anthrax is enzootic in many Asian and African countries, and is reported in Australia, some parts of Europe, and America. The clinical courses of this disease in animals are peracute, acute, subacute, and chronic forms. In severely infected cases, the animals are dead without premonitory clinical signs. The blood may fail to clot and can be found in the mouth, nostrils, and anus in the animals that die from anthrax. This bacterium is susceptible to many antibiotics, yet only penicillin and oxytetracycline have the most effective under field conditions. When an outbreak occurs in a defined area, it is necessary to take early steps to break the infection cycle by maintaining strict biosecurity and vaccinating uninfected animals. This disease is still a challenge to farm animal production in many countries. This review intends to give a fair knowledge of the etiology, epidemiology, pathogenesis, clinical presentation, diagnosis, treatment, and control of this disease.

Effects of the preservation medium and storage duration of domestic cat ovaries on the maturational and developmental competence of oocytes in vitro.

We examined the effectiveness of saline, Euro-Collins solution (EC), and ET-Kyoto solution (ET-K) as preservation media for the cold storage of feline ovaries. Ovaries were maintained in these media at 4°C for 24, 48, or 72 h until oocyte retrieval. The ET-K group exhibited a higher oocyte maturation rate than the saline group after 72 h of storage. Moreover, ET-K could sustain the competence of the feline oocytes to cleave after 48 h, and the morula formation rate of the ET-K group was higher than that of the other groups after 24 and 48 h. Furthermore, the ET-K group exhibited a higher blastocyst formation rate than the other groups after storage for 24 h, and only ET-K retained the developmental competence in blastocysts after 48 h of storage. In addition, regarding the cell numbers of the blastocysts, there was no significant difference among the tested groups. In conclusion, our results indicate that ET-K is a suitable preservation medium for feline ovaries.

Effects of tris (hydroxymethyl) aminomethane and egg yolk on the cryopreservation of buck semen.

Objectives: This study was designed to examine the effects of various concentrations of tris (hydroxymethyl) aminomethane (tris) and egg yolk on the quality of cryopreserved buck sperm. Materials and Methods: The collected semen samples were pooled, washed, and diluted into five different freezing extender groups, viz., extender I (tris 0% + egg yolk 0%), extender II (tris 1.41% + egg yolk 4%), extender III (tris 2.41% + egg yolk 8%), extender IV (tris 3.41% + egg yolk 16%), and extender V (tris 4.41% + egg yolk 24%). The sperm parameter of the five groups of extenders was evaluated after equilibration and cryopreservation. Results: The results showed that extenders II-V provided significantly higher semen progressive motility and total motility percentages than extender I after equilibration (p < 0.05). The higher percentages of semen progressive motility, total motility, viability, and plasma membrane integrity (by both HOST under light microscopy and stain after HOST under light microscopy) were found in the sperm cryopreserved with extender IV than extender I, extender II, and extender III groups after thawing (p < 0.05). In addition, semen progressive motility, total motility, and viability were not further increased, or plasma membrane integrity (by both HOST tests) was decreased by the addition of tris and egg yolk (extender V) after cryopreservation (p < 0.05). Conclusion: In conclusion, our result indicates that the following washing, the supplementation of tris (3.41% + egg yolk 16%) on the freezing extender are suitable for improving the semen quality of buck after freezing and thawing.

Feline embryo development in commercially available human media supplemented with fetal bovine serum.

Feline embryo development was examined for 7 days after fertilization using commercially available human media supplemented with 0.3% bovine serum albumin (BSA) or 5% fetal bovine serum (FBS). Cumulus-oocyte complexes were categorized as Grades 1, 2, and 3 according to morphology. Only-One Medium (OM) was used for in vitro culture (IVC) in OM + BSA, OM + FBS, and OM + BSA/FBS, with BSA supplementation for the first 2 days and FBS for the subsequent 5 days. Embryos cultured in Early Culture Medium (1-2 days) and Blastocyst Medium (3-7 days) were defined as EB + BSA and EB + BSA/FBS. The developmental rate until the blastocyst stage of Grade 1 and 2 oocytes cultured in OM + BSA/FBS was higher than for the other groups and was significantly higher than for the OM + BSA and EB + BSA groups (P<0.01). Grade 3 oocytes cultured in OM + BSA/FBS also showed the greatest proportion of blastocyst formation. However, FBS supplementation throughout the IVC period reduced blastocyst number. The percentage of 2 pronuclei after fertilization as well as blastocyst cell number were significantly higher in Grade 1 and 2 than Grade 3 oocytes when cultured in OM + BSA/FBS (P<0.05). These results indicate that commercially available OM supplemented with BSA for the first 2 days of culture and FBS for the subsequent 5 days is suitable for feline embryo development until the blastocyst stage.

Development of feline embryos produced by Piezo-actuated intracytoplasmic sperm injection of elongated spermatids.

Piezo-actuated intracytoplasmic sperm injection (Piezo-ICSI) is used as an efficient in vitro fertilization method with various animals. With this method, elongated spermatids are collected from testicular tissues and are easier to obtain from animals that unexpectedly die than ejaculate sperm. Additionally, elongated spermatid injection often results in the development of embryos and offspring. To develop assisted reproductive techniques (ARTs) for domestic cats, we examined the effects of oocyte activation on cleavage and embryo development after Piezo-ICSI with motile sperm (experiment 1) and after Piezo-ICSI with either testicular sperm or elongated spermatids (experiment 2). In experiment 1, the proportions of cleaved embryos, morulas, and blastocysts following Piezo-ICSI with ethanol activation were significantly higher (P < 0.05) than in the non-activated groups. However, the proportion of blastocysts and the blastocyst quality did not differ significantly (P > 0.05) between the ethanol-activated and non-activated groups. In experiment 2, the cleavage frequencies of oocytes after Piezo-ICSI of testicular sperm or elongated spermatids and ethanol activation were higher (P < 0.05) than that of oocytes in the non-activated group, but the occurrence of blastocyst formation and quality of blastocysts did not differ between the activated and non-activated groups. In summary, cat embryos can be produced by Piezo-actuated microinjection of elongated spermatids. Ethanol activation increased the frequency of cleavage, but it affected neither the occurrence of blastocyst development nor the quality of blastocysts. These results represent an expansion in the repertoire of ARTs that are potentially applicable to both domestic and endangered species of cats.

Generation of Footprint-Free Canine Induced Pluripotent Stem Cells Using Auto-Erasable Sendai Virus Vector.

Canine induced pluripotent stem cells (ciPSCs) can be used in regenerative medicine. However, there are no reports on the generation of genome integration-free and completely exogenous gene-silenced (footprint free) ciPSCs that are tolerant to enzymatic single-cell passage. In this study, we reprogrammed canine embryonic fibroblasts using the auto-erasable replication-defective and persistent Sendai virus vector, SeVdp(KOSM)302L, and generated two ciPSC lines. The ciPSCs were positive for pluripotent markers, including alkaline phosphatase activity as well as OCT3/4, SOX2, and NANOG transcripts, and NANOG, stage-specific embryonic antigen-1, and partial TRA-1-60 protein expression, even after SeVdp(KOSM)302L removal. The ciPSCs were induced to differentiate into all the three germ layers as embryoid bodies in vitro and as teratomas in vivo. Furthermore, SeVdp(KOSM)302L-free ciPSCs maintained a normal karyotype even after repeated enzymatic single-cell passaging. Therefore, to our knowledge, for the first time, we demonstrated the generation of footprint-free and high-quality ciPSCs that can be passaged at the single-cell stage using enzymatic methods. Our method for generation of ciPSCs is a good step toward the development of clinical application of ciPSCs.