Background: The small intestine plays a crucial role in animals in maintaining homeostasis as well as a series of physiological events such as nutrient uptake and immune function to improve productivity. Research on intestinal organoids has recently garnered interest, aiming to study various functions of the intestinal epithelium as a potential alternative to an in vivo system. These technologies have created new possibilities and opportunities for substituting animals for testing with an in vitro model. Methods: Here, we report the establishment and characterisation of intestinal organoids derived from jejunum tissues of adult pigs. Intestinal crypts, including intestinal stem cells from the jejunum tissue of adult pigs (10 months old), were sequentially isolated and cultivated over several passages without losing their proliferation and differentiation using the scaffold-based and three-dimensional method, which indicated the recapitulating capacity. Results: Porcine jejunum-derived intestinal organoids showed the specific expression of several genes related to intestinal stem cells and the epithelium. Furthermore, they showed high permeability when exposed to FITC-dextran 4 kDa, representing a barrier function similar to that of in vivo tissues. Collectively, these results demonstrate the efficient cultivation and characteristics of porcine jejunum-derived intestinal organoids. Conclusions: In this study, using a 3D culture system, we successfully established porcine jejunum-derived intestinal organoids. They show potential for various applications, such as for nutrient absorption as an in vitro model of the intestinal epithelium fused with organ-on-a-chip technology to improve productivity in animal biotechnology in future studies.

Background: Somatic cell nuclear transfer (SCNT) is a prominent technology that can preserve superior genetic traits of animals and expand the population in a short time. Hematological characters and endocrine profiles are important elements that demonstrate the stability of the physiological state of cloned animals. To date, several studies regarding cloned camels with superior genes have been conducted. However, detailed hemato-physiological assessments to prove that cloned camels are physiologically normal are limited. In this study, We evaluated the hemato-physiological characteristics of cloned male and female dromedary camels (Camelus dromedaries). Methods: Therefore, we analyzed variations in hematological characteristics and endocrine profiles between cloned and non-cloned age-matched male and female dromedary camels (Camelus dromedaries). Two groups each of male and female cloned and non-cloned camels were monitored to investigate the differences in hemato-physiological characteristics. Results: All the animals were evaluated by performing complete blood count (CBC), serum chemistry, and endocrinological tests. We found no significant difference between the cloned and non-cloned camels. Furthermore, the blood chemistry and endocrine profiles in male and female camels before maturity were similar. Conclusions: These results suggest that cloned and non-cloned camels have similar hematological characteristics and endocrine parameters.

Background: Although an understanding of the proliferation and differentiation of fish female germline stem cells (GSCs) is very important, an appropriate threedimensional (3D) research model to study them is not well established. As a part of the development of stable 3D culture system for fish female GSCs, we conducted this study to establish a 3D aggregate culture system of ovarian cells in marine medaka, Oryzias dancena. Methods: Ovarian cells were separated by Percoll density gradient centrifugation and two different cell populations were cultured in suspension to form ovarian cell aggregates to find suitable cell populations for its formation. Ovarian cell aggregates formed from different cell populations were evaluated by histology and gene expression analyses. To evaluate the media supplements, ovarian cell aggregate culture was performed under different media conditions, and the morphology, viability, size, gene expression, histology, and E2 secretion of ovarian cell aggregates were analyzed. Results: Ovarian cell aggregates were able to be formed well under specific culture conditions that used ultra-low attachment 96 well plate, complete mESM2, and the cell populations from top to 50% layers after separation of ovarian cells. Moreover, they were able to maintain minimal ovarian function such as germ cell maintenance and E2 synthesis for a short period. Conclusions: We established basic conditions for the culture of O. dancena ovarian cell aggregates. Additional efforts will be required to further optimize the culture conditions so that the ovarian cell aggregates can retain the improved ovarian functions for a longer period of time.

Background: The biological information of fish, which include reproduction, is the prerequisite and the basis for the assessment of fisheries. Methods: The aim of this work was to know the reproductive biology with the first sexual maturity (TL₅₀) and the spawning period for 58 mainly fish species in the waters around La Réunion Island (Western Indian Ocean). Twenty families belonging to the Actinopterygii were represented (acanthuridae, berycidae, bramidae, carangidae, cirrhitidae, gempylidae, holocentridae, kyphosidae, labridae, lethrinidae, lutjanidae, malacanthidae, monacanthidae, mullidae, polymixiidae, pomacentridae, scaridae, scorpaenidae, serranidae, sparidae; 56 species; n = 9,751) and two families belonging to the Elasmobranchii (squalidae, centrophoridae; 2 species; n = 781) were sampled. Between 2014 and 2022, 10,532 individuals were sampled covering the maximum months number to follow the reproduction periods of these species. Results: TL₅₀ for the males and the females, respectively, ranged from 103.9 cm (Acanthurus triostegus) to 1,119.3 cm (Thyrsitoides marleyi) and from 111.7 cm (A. triostegus) to 613.1 cm (Centrophorus moluccensis). The reproduction period could be very different between the species from the very tight peak to a large peak covered all months. Conclusions: Most species breed between October and March but it was not the trend for all species around La Réunion Island.

Background: Sperm quality and the number of sperm introduced into the uterus during artificial insemination (AI) are pivotal factors influencing pregnancy outcomes. However, there have been no reports on the relationship between sperm concentration at AI and sperm quality in Hanwoo cattle. In this study, we examined sperm quality and pregnancy rates after AI using sperm inseminated at different concentrations. Methods: We evaluated the motility, viability, and acrosomal membrane integrity of sperm at different concentrations (10, 15, 18, and 20 million sperm/straw) in 0.5-mL straws. Subsequently, we compared the pregnancy rates after AI with different sperm concentrations. Results: After freeze-thawing, sperm at the assessed concentrations showed similar viability and acrosomal membrane integrity. After AI, cattle in the 10 million group had significantly lower pregnancy rates compared to those in the 18 and 20 million groups. Conversely, there were no statistically significant variances observed between cattle in the 10 and 15 million groups. Conclusions: Sperm at concentrations of 10, 15, 18 and 20 million per straw exhibited comparable motility, viability, and acrosomal membrane integrity. However, a concentration of at least 18 million sperm per straw is required to achieve a consistent rate of pregnancy rate in Hanwoo cattle after AI.

Background: Cadmium (Cd) is toxic heavy metal that accumulates in organisms after passing through their respiratory and digestive tracts. Although several studies have reported the toxic effects of Cd exposure on human health, its role in embryonic development during preimplantation stage remains unclear. We investigated the effects of Cd on porcine embryonic development and elucidated the mechanism. Methods: We cultured parthenogenetic embryos in media treated with 0, 20, 40, or 60 µM Cd for 6 days and evaluated the rates of cleavage and blastocyst formation. To investigate the mechanism of Cd toxicity, we examined intracellular reactive oxygen species (ROS) and glutathione (GSH) levels. Moreover, we examined mitochondrial content, membrane potential, and ROS. Results: Cleavage and blastocyst formation rates began to decrease significantly in the 40 µM Cd group compared with the control. During post-blastulation, development was significantly delayed in the Cd group. Cd exposure significantly decreased cell number and increased apoptosis rate compared with the control. Embryos exposed to Cd had significantly higher ROS and lower GSH levels, as well as lower expression of antioxidant enzymes, compared with the control. Moreover, embryos exposed to Cd exhibited a significant decrease in mitochondrial content, mitochondrial membrane potential, and expression of mitochondrial genes and an increase in mitochondrial ROS compared to the control. Conclusions: We demonstrated that Cd exposure impairs porcine embryonic development by inducing oxidative stress and mitochondrial dysfunction. Our findings provide insights into the toxicity of Cd exposure on mammalian embryonic development and highlight the importance of preventing Cd pollution.

Three different dogs who had immune-mediated hemolytic anemia (IMHA) were treated for more than two weeks with blood transfusion in an animal clinic. Despite this treatment and hospitalization, there was no clinical improvement in clinical signs as well as complete blood cell count (CBC) including hematocrit (HCT) and C-reactive protein (CRP). All cases were then injected two or three times with allogeneic stem cells through an intravenous route for treatment. Upon administrating stem cells to the IMHA dogs, clinical conditions and the indexes of HCT and CRP were clinically improved within or close to normal ranges.

Dystocia, a challenging condition in obstetrics, can arise from various causes, including fetal monsters with structural abnormalities. This case report presents a unique case of dystocia due to a fetal monster known as Perosomus Elumbis in a beetal breed goat from Pakistan. The 4-years-old pregnant doe presented with prolonged straining and failure to deliver the fetus after 8 hours of labor. Upon examination, the cervix was dilated, and only the forelimbs of the fetus were visible in the birth canal. The subsequent delivery involved the application of manual traction by using a dystocia kit, and the removal of edematous fluid from the legs. The monster fetus exhibited absence of hair growth, along with the absence of thoracic vertebrae. Two other fetuses were present, with one found dead and the other alive. Posttreatment involved fluid therapy, antibiotics, and supportive care for the doe. This case report sheds light on the occurrence of Perosomus Elumbis fetal monsters and their impact on dystocia in goat breeding. Understanding the underlying causes and implementing appropriate management strategies are crucial for successful outcomes in similar cases.