AhR Activation Ameliorates Intestinal Barrier Damage in Immunostressed Piglets by Regulating Intestinal Flora and Its Metabolism.

The primary factor leading to elevated rates of diarrhea and decreased performance in piglets is immunological stress. The regulation of immune stress through the intestinal flora is a crucial mechanism to consider. In total, 30 weaned piglets were randomly allocated to five groups: the basal diet group (Control), basal diet + lipopolysaccharides group (LPS), basal diet + 250 µg/kg 6-Formylindolo [3,2-b] carbazole + LPS group (FICZ), basal diet + 3mg/kg Cardamonin + LPS group (LCDN), and basal diet + 6mg/kg Cardamonin + LPS group (HCDN/CDN). The results showed that compared with those of the LPS group, the expression of tight junction proteins (occludin; claudin-1) in the FICZ group was significantly increased, and the mRNA levels of IL-1ß and TNF-α were significantly reduced (p < 0.05). HCDN treatment had a better effect on LPS-induced intestinal barrier damage in this group than it did in the LCDN group. HCDN treatment leads to a higher villus height (VH), a higher ratio of villi height to crypt depth (V/C), higher tight junction proteins (ZO-1; occludin), and higher short-chain fatty acids (SCFAs). In addition, correlation analyses showed that Succinivibrio was positively correlated with several SCFAs and negatively correlated with prostaglandin-related derivatives in the FICZ group and CDN group (p < 0.05). In summary, Cardamonin alleviates intestinal mucosal barrier damage and inflammatory responses by regulating the intestinal microbiota and its metabolism.

Exploring the Rumen Microbiota and Serum Metabolite Profile of Hainan Black Goats with Different Body Weights before Weaning.

The critical role of the rumen microbiota in the growth performance of livestock is recognized, yet its significance in determining the body weight of goat kids before weaning remains less understood. To bridge this gap, our study delved into the rumen microbiota, serum metabolome, rumen fermentation, and rumen development in goat kids with contrasting body weights before weaning. We selected 10 goat kids from a cohort of 100, categorized into low body weight (LBW, 5.56 ± 0.98 kg) and high body weight (HBW, 9.51 ± 1.01 kg) groups. The study involved sampling rumen contents, tissues, and serum from these animals. Our findings showed that the HBW goat kids showed significant enrichment of VFA-producing bacteria, particularly microbiota taxa within the Prevotellaceae genera (UCG-001, UCG-003, and UCG-004) and the Prevotella genus. This enrichment correlated with elevated acetate and butyrate levels, positively influencing rumen papillae development. Additionally, it was associated with elevated serum levels of glucose, total cholesterol, and triglycerides. The serum metabonomic analysis revealed marked differences in fatty acid metabolism between the LBW and HBW groups, particularly in encompassing oleic acid and both long-chain saturated and polyunsaturated fatty acids. Further correlational analysis underscored a significant positive association between Prevotellaceae_UCG-001 and specific lipids, such as phosphatidylcholine (PC) (22:5/18:3) and PC (20:3/20:1) (r > 0.60, p < 0.05). In summary, this study underscores the pivotal role of the rumen microbiota in goat kids' weight and its correlation with specific serum metabolites. These insights could pave the way for innovative strategies aimed at improving animal body weight through targeted modulation of the rumen microbiota.

Dietary emodin alleviates lipopolysaccharide-induced intestinal mucosal barrier injury by regulating gut microbiota in piglets.

This study was to determine the effects of dietary emodin (ED) on the intestinal mucosal barrier, nuclear factor kappa-B (NF-κB) pathways, and gut microbial flora in lipopolysaccharide (LPS)-induced piglets. Twenty-four weaned piglets were chosen and 4 treatments were created by randomly distributing piglets into CON, ED, LPS, and ED_LPS groups. Experiments were done in a 2 × 2 factorial arrangement and maintained for 21 d. Dietary treatment (a basal diet or 300 mg/kg ED) and immunological challenge (LPS or sterile saline) were 2 major factors. Intraperitoneal injections of LPS or sterilized saline were given to piglets on d 21. Six hours after the LPS challenge, all piglets were euthanized for sample collection and analysis. The results showed that piglets of the ED_LPS group had higher (P < 0.05) villus height to crypt depth ratio (VCR), and lower (P < 0.05) plasma D-lactate and diamine oxidase (DAO) than the LPS group. Furthermore, ED inhibited (P < 0.05) the decrease of glutathione peroxidase (GSH-Px) and catalase (CAT) activities and increase of malonaldehyde level (P < 0.05) in jejunal mucosa induced by LPS. The mRNA levels of pro-inflammatory cytokine genes (IL-6, IL-1ß, and TNF-α) were significantly reduced (P < 0.05), and the mRNA levels of antioxidant enzyme genes (GPX-1, SOD2 and CAT), as well as protein and mRNA levels of tight junction proteins (occludin, claudin-1, and ZO-1), were also significantly increased (P < 0.05) by ED addition in LPS-induced piglets. Meanwhile, ED supplementation significantly decreased the LPS-induced protein levels of cyclooxygenase-2 and phosphorylation levels of NF-κB p65 and IκBα in jejunal mucosa. Emodin had a significant effect on the composition of gut microbial flora at various taxonomic positions as indicated by 16S RNA sequencing. The acetic acid, isobutyric acid, valeric acid, and isovaleric acid concentrations in the cecum were also increased by ED addition in pigs (P < 0.05). Furthermore, the correlation analysis revealed that some intestinal microbiota had a potential relationship with jejunal VCR, plasma D-lactate and DAO, jejunal mucosa GSH-Px and CAT activity, and cecal short-chain fatty acid concentration. These data suggest that ED is effective in alleviating LPS-induced intestinal mucosal barrier injury by modulating gut microbiota in piglets.

Resveratrol Attenuates Diquat-Induced Oxidative Stress by Regulating Gut Microbiota and Metabolome Characteristics in Piglets.

Previous studies have shown that dietary resveratrol (RES) reduces diarrhea and attenuates oxidative stress in piglets challenged with diquat. However, the effect of dietary resveratrol on the gut microbiota of these piglets, as well as the potential relationships between intestinal microflora and metabolites, remain unclear. Here, 16S ribosomal DNA sequencing and metabolome analyses were performed to investigate the effect of RES on the gut microbiota and metabolome of diquat-challenged piglets. A total of 18 weaned piglets (aged 28 ± 2 days) were divided into the control group (basal diet), diquat group (basal diet + diquat challenge), and RES group (basal diet containing 90 mg/kg RES + diquat challenge). Compared with the control group, piglets in the diquat group showed enriched relative abundance of the phyla Firmicutes and Actinobacteria, the genus Ruminococcaceae UCG-005, and members of the Eubacterium coprostanoligenes group. Noteworthy, RES supplementation significantly reduced the levels of these microorganisms. In contrast, the relative abundance of some beneficial bacterial species in the RES group, such as the genera Clostridium sensu stricto 1 and Lachnospiraceae unclassified were significantly higher than in the diquat and control groups. Metabolomic analysis indicated that some metabolites, including indole-3-carbinol, 5-hydroxyindole-3-acetic acid, and uridine, were significantly upregulated upon RES supplementation. In particular, the relative abundance of uridine, indole, and alpha- and beta-dihydroresveratrol was significantly higher in the RES group than in the control group. Moreover, most gut bacterial genera were found to be highly correlated with altered gut microbiota-related metabolites. These findings suggest that dietary supplementation with resveratrol may alter the composition and metabolites of colonic microbiota in diquat-challenged piglets, which provides important insights into the use of resveratrol as a feed additive for gut microbial regulation in piglets with inflammatory and oxidative stress-associated disorders.

Resveratrol protects intestinal integrity, alleviates intestinal inflammation and oxidative stress by modulating AhR/Nrf2 pathways in weaned piglets challenged with diquat.

This study investigated the effects of resveratrol (RES) on intestinal morphology, antioxidant capacity, intestinal inflammation, and barrier function in weaned piglets challenged with diquat (DIQ). Thirty weaned piglets were randomly assigned to 5 treatments: non-challenged group (CON), DIQ-challenged group (DIQ), and DIQ-challenged group with 10, 30, or 90 mg/kg of RES, respectively. The trail lasted 21 days, and piglets were intraperitoneally injected with DIQ or the same amount of saline on day 15. The results showed that supplementation with 90 mg/kg RES increased (P < 0.05) jejunal villus height and villus height: crypt depth ratio, and decreased (P < 0.05) crypt depth, plasma D-lactate and diamine oxidase (DAO) compared with the DIQ group. Piglets fed with 30 or 90 mg/kg RES prevented the diquat-induced decrease (P < 0.05) of mRNA expression of occludin, claudin-1, ZO-1, and IL-10, and increase (P < 0.05) of TNF-α mRNA expression. Moreover, addition of 90 mg/kg RES increased (P < 0.05) the activities of SOD, GSH-Px, and CAT and decreased (P < 0.05) the MDA levels in jejunal mucosa compared with the DIQ group. Finally, addition of 90 mg/kg RES enhanced (P < 0.05) the mRNA expression of SOD1, SOD2, CAT, GPx1, and HO-1, and increased (P < 0.05) mRNA and protein expression of Nrf2, NQO1, aryl hydrocarbon receptor (AhR), and cytochrome P450 family 1 member A1 (CYP1A1). These data indicated that supplementation with 90 mg/kg RES was effective in protecting the intestinal integrity, alleviating intestinal inflammation and oxidative stress by activating AhR/Nrf2 pathways in diquat-challenged piglets.

Effect of the Single and Combined Use of Curcumin and Piperine on Growth Performance, Intestinal Barrier Function, and Antioxidant Capacity of Weaned Wuzhishan Piglets.

This study was conducted to evaluate effects of the single and combined use of curcumin (CUR) and piperine (PIP) on performance, intestinal barrier function, and antioxidant capacity of weaned piglets. A total of 50 Wuzhishan piglets weaned at 35 days of age were randomly assigned to five groups receiving a corn-soybean basal diet (CON), the basal diet supplemented with 50 mg/kg piperine, 200 mg/kg curcumin (low-CUR), 200 mg/kg curcumin + 50 mg/kg piperine (PIP + CUR), and 300 mg/kg curcumin (high-CUR), respectively. The results showed that the feed/gain ratio (F/G) and plasma d-lactate and diamine oxidase activity (DAO) of the CUR + PIP and high-CUR groups were lower than those of the CON group (all P < 0.05), while the jejunum and ileum villus height, the villus height/crypt depth ratio, and the messenger RNA (mRNA) expression levels of occludin, claudin-1, and zonula occluden-1 in jejunal and ileal mucosa were higher in the CUR + PIP and high-CUR groups than in the CON group (all P < 0.05). Moreover, the piglets in the CUR + PIP and high-CUR groups had higher serum and intestinal mucosa activity of superoxide dismutase and glutathione peroxidase and lower malonaldehyde concentration than piglets in the CON group (all P < 0.05). The above parameters were not significantly different between the CUR + PIP and high-CUR groups (P > 0.05). In conclusion, the combination of CUR and PIP seemed to be as advantageous as high-CUR to piglets, but it was more effective than the single use of CUR and PIP. These data indicated that the basal diet supplemented with CUR + PIP or high-CUR could improve the intestinal permeability and suppress oxidative stress of weaned Wuzhishan piglets.

Moderate Coconut Oil Supplement Ameliorates Growth Performance and Ruminal Fermentation in Hainan Black Goat Kids.

The study investigated amelioration effects of coconut oil (CO) on growth performance, nutrient digestibility, ruminal fermentation, and blood metabolites in Hainan Black goat kids. Twenty-four Hainan Black goat kids (10 days of age) were assigned randomly to four treatments for 90 days, including pre-weaning (10-70 d of age) and post-weaning (70-100 d of age) days. The treatment regimens were control (CON), low CO (LCO), medium CO (MCO), and high CO (HCO) with 0, 4, 6, 8 g CO per goat per day, respectively. During the pre-weaning period, the average daily gain (ADG) linearly and quadratically increased (P < 0.05), whereas the average daily feed intake (ADFI) linearly decreased, and the feed conversion ratio (FCR) also decreased linearly and quadratically by increasing CO supplementation (P < 0.05). During the post-weaning period, increasing CO supplementation linearly and quadratically increased the BW at 100 days and ADG (P < 0.05), but quadratically decreased the ADFI and FCR (P < 0.05). The digestibility of ether extract (EE) linearly and quadratically increased with increasing CO supplementation (P < 0.05). Supplementation of CO linearly increased ruminal pH (P < 0.05), but linearly decreased (P < 0.05) ammonia-N, total VFAs, molar proportions of acetate, ruminal microbial enzyme activity of carboxymethyl-cellulase, cellobiase, xylanase, pectinase and α-amylase, and number of total protozoa, the abundance of Ruminococcus albus, Ruminococcus flavefaciens, Fibrobacter succinogenes, Butyrivibrio fibrisolvens, Prevotella ruminicola, and Ruminobacter amylophilus. The estimated methane emission decreased linearly and quadratically with increasing CO addition (P < 0.05). The serum concentration of triglycerides (TG), non-esterified fatty acids (NEFA) and growth hormone (GH) linearly (P < 0.05) increased by raising the CO supplementation. The present results indicate that CO supplementation at 6 g/day per goats is optimum due to improved growth performance and decreased estimated methane emission. Supplementation CO up to 8 g/day depressed growth and feed conversion due to its suppression of growth performance, rumen protozoa, cellulolytic bacteria and microbial enzyme activity, and reduced ADF and ADF digestibility.

Landscape of alternative splicing in Capra_hircus.

Alternative splicing (AS) is a fundamental regulatory process in all higher eukaryotes. However, AS landscapes for a number of animals, including goats, have not been explored to date. Here, we sequenced 60 samples representing 5 tissues from 4 developmental stages in triplicate using RNA-seq to elucidate the goat AS landscape. In total, 14,521 genes underwent AS (AS genes), accounting for 85.53% of intron-containing genes (16,697). Among these AS genes, 6,342 were differentially expressed in different tissues. Of the AS events identified, retained introns were most prevalent (37.04% of total AS events). Functional enrichment analysis of differential and specific AS genes indicated goat AS mainly involved in organ function and development. Particularly, AS genes identified in leg muscle were associated with the "regulation of skeletal muscle tissue development" GO term. Given genes were associated with this term, four of which (NRG4, IP6K3, AMPD1, and DYSF) might play crucial roles in skeletal muscle development. Further investigation indicated these five genes, harbored 13 ASs, spliced exclusively in leg muscle, likely played a role in goat leg muscle development. These results provide novel insights into goat AS landscapes and a valuable resource for investigation of goat transcriptome complexity and gene regulation.

Effects of curcumin on growth performance, jejunal mucosal membrane integrity, morphology and immune status in weaned piglets challenged with enterotoxigenic Escherichia coli.

The aim of this study was to evaluate the effect of dietary curcumin supplementation on growth performance and intestinal mucosal barrier function of weaned piglets. Fifty piglets, weaned at 21±2 days of age, were randomly allotted to five treatments for 21 days. The dietary treatments were the control (basal diet), and the basal diet supplemented with 50mg/kg quinocetone, or 200 mg/kg, 300 mg/kg or 400mg/kg curcumin. The piglets were housed in individual pens and orally challenged with enterotoxigenic Escherichia coli (ETEC) during the preliminary trial period. The jejunal morphology and histology analysis were detected under light microscope. The plasma D-lactate and diamine oxidase (DAO) were determined by using enzymatic spectrophotometric assay. Immunohistochemistry assays were used to examine secretory immunoglobulin (sIgA) protein expression. Real-time PCR was used to determine mRNA levels of cytokine and Toll-like receptor 4 (TLR4) in jejunal mucosa. The results showed that, compared with the control, dietary addition of 300 mg/kg or 400 mg/kg curcumin decreased (P<0.05) feed/gain ratio and crypt depth, improved (P<0.05) villus height and villus height:crypt depth ratio, reduced (P<0.05) plasma D-lactate and DAO activity, up-regulated the protein expression of sIgA (P<0.05), increased (P<0.05) the number of goblet cells (GCs) and reduced (P<0.05) the number of intraepithelial lymphocytes (IELs). The mRNA levels of interleukin 1ß (IL-1ß) and TLR4 and tumor necrosis factor α (TNF-α) were also decreased (P<0.05), but mRNA level of interleukin 10 (IL-10) was increased (P<0.05). There was no difference in the above parameters between the 300 mg/kg and 400 mg/kg curcumin groups. Pigs fed with 50 mg/kg quinocetone also decreased (P<0.05) feed/gain ratio, increased villus height:crypt depth ratio (P<0.05), and reduced (P<0.05) crypt depth and mRNA levels of TLR4. In conclusion, curcumin and the quinocetone have similar effects in improving piglet growth, but dietary addition of 300 mg/kg or 400 mg/kg curcumin was more effective than quinocetone in improving intestinal mucosal barrier integrity, morphology, and immune status of weaned pigs. This indicates that curcumin could be used as a potential feed additive replacing quinocetone in weaned piglets.

Dual functions in response to heat stress and spermatogenesis: characterization of expression profile of small heat shock proteins 9 and 10 in goat testis.

Small heat shock proteins 9 and 10 (HSPB9 and HSPB10) are two testis-specific expressed sHsps. The objective of this study was to investigate the mRNA expression profile of HSPB9 and HSPB10 in goat testis among the different seasons, ages, and environmental temperatures. Allocation of the two sHsps was also performed by immunohistochemistry. The results showed that the transcript levels of HSPB9 and HSPB10 were extremely high in the testis (P < 0.01). The relative expression of HSBP9 and HSPB10 in testis showed a tendency to increase with age and then is maintained at the constant level after sexual maturity. HSPB9 and HSPB10 have significantly higher expression in the breeding season (P < 0.05) and hot season (P < 0.01). Both HSPB9 and HSPB10 were found to be upregulated by high-temperature stress in testis (P < 0.05), and the expressions of Hsp70 and Hsp90 were also increased simultaneously (P < 0.01). Immunohistochemistry analysis localized HSPB9 expressed in spermatogonia, spermatocytes, and round spermatids and HSPB10 expressed in the elongate spermatids. In epididymis, strongly staining signal of HSPB10 was detected in pseudostratified columnar epithelium. In conclusion, the two testis-specific sHsps are closely related to male reproduction and heat tolerance. The results could provide valuable data for the further studies on HSPB9 and HSPB10.

Effect of rapid freezing-thawing techniques on the sperm parameters and ultrastructure of Chinese Taihang black goat spermatozoa.

Supercooling sperm in liquid nitrogen vapour is a feasible and economic technique for the practical production. The study aimed to reveal the negative effects of this rapid freezing and thawing processes on Taihang black goat spermatozoa and to find out the changing of spermatozoa motility and ultrastructure by using CASA and TEM. Qualified semen samples, which collected from twenty Chinese Taihang black goats using artificial vagina were pooled and investigated the kinematics parameters and ultrastructural morphology. The results showed that freezing-thawing caused a significant reduction in the spermatozoon total motility (P<0.001), in rapid and medium cell numbers (P<0.001) and motility parameters (VAP, VSL, VCL, ALH and BCF) (P<0.01). Immotile spermatozoa number was increased significantly after freezing-thawing (P<0.001). In the ultrastructural analysis, the shape with a sperm nucleus characterized by ruptures, bend and deformity was observed. The plasma membranes were broken, and nucleoplasm erupted. The mitochondria in the middle piece were disturbed by partial absence or additional accumulations. Swelling, coiling, vacuolization and structural disorganization of mitochondria were also observed. In conclusion, Freezing-thawing procedure has a detrimental effect on motility, membrane integrity and mitochondria of goat spermatozoa. Transmission electron microscopy provides an intuitive observation to investigate deformity spermatozoa.

Ultrastructure of germ cells, Sertoli cells and mitochondria during spermatogenesis in mature testis of the Chinese Taihang black goats (Capra hircus).

The objective of this study was to describe the ultrastructure of germ cells, Sertoli cells and mitochondria in mature testis of the Chinese Taihang black goat. The characteristics of germ cell nucleus and mitochondria changing during spermatogenesis were investigated by transmission electron microscopy (TEM). The results showed that the spermatogonium was elliptical, and its nucleus was about 4-5 µm. The round mitochondria can be observed throughout the cytoplasm around the nucleus. Small patches of heterochromatin were distributed throughout the nucleus. Spermatocyte was oval-shaped with a nucleus of about 4-4.5 µm in diameter. The heterochromatin began to attach to the inner surface of the nuclear membrane. Spermatid was about 4 µm and oval in shape. Its nucleus was oval or round and approximately 2-3 µm in diameter. The borderline between nucleus membrane and karyoplasm was distinct. During spermiogenesis, spermatid nucleus was condensed and elongated, and chromatin reached the highest condensation in the mature spermatozoon. The mid-piece was surrounded by mitochondria at the neck region. The sperm tail showed the typical "9+2″ structure, contained axoneme and central singlet microtubules. The nuclei of the Sertoli cells were irregular shaped and showed indentations in the membrane. In the mature testes of goat bucks, abundant mitochondria were around the germ cells and Sertoli cells. The scattered mitochondria were aggregated around the base of the flagellum (axoneme) during the spermatid differentiation stage. In conclusion, the present study showed that the spermatogenic process of Taihang black goat followed the pattern of mammals with some specific.

Effect of high-dose nano-selenium and selenium-yeast on feed digestibility, rumen fermentation, and purine derivatives in sheep.

The aim of this study was to evaluate the effect of nano-selenium (NS) and yeast-selenium (YS) supplementation on feed digestibility, rumen fermentation, and urinary purine derivatives in sheep. Six male ruminally cannulated sheep, average 43.32 ± 4.8 kg of BW, were used in a replicated 3 × 3 Latin square experiment. The treatments were control (without NS and YS), NS with 4 g nano-Se (provide 4 mg Se), and YS with 4 g Se-yeast (provide 4 mg Se) per kilogram of diet dry matter (DM), respectively. Experimental periods were 25 days with 15 days of adaptation and 10 days of sampling. Ruminal pH, ammonia N concentration, molar proportion of propionate, and ratio of acetate to propionate were decreased (P < 0.01), and total ruminal VFA concentration was increased with NS and YS supplementation (P < 0.01). In situ ruminal neutral detergent fiber (aNDF) degradation of Leymus chinensis (P < 0.01) and crude protein (CP) of soybean meal (P < 0.01) were significantly improved by Se supplementation. Digestibilities of DM, organic matter, crude protein, ether extract, aNDF, and ADF in the total tract and urinary excretion of purine derivatives were also affected by feeding Se supplementation diets (P < 0.01). Ruminal fermentation was improved by feeding NS, and feed conversion efficiency was also increased compared with YS (P < 0.01). We concluded that nano-Se can be used as a preferentially available selenium source in ruminant nutrition.

Cloning, characterization, and expression analysis of goat (Capra hircus) phospholipid hydroperoxide glutathione peroxidase (PHGPx).

Phospholipid hydroperoxide glutathione peroxidase (PHGPx), as a ubiquitous antioxidant enzyme in the glutathione peroxidases (GPx) family, plays multiple roles in organisms. However, there is very little information on PHGPx in goats (Capra hircus). In this study, a full-length cDNA was cloned and characterized from Taihang black goat testes. The 844 bp cDNA contains an open reading frame (ORF) of 597 bp. The goat PHGPx nucleotide sequence contains a selenocysteine (sec) codon TGA(244-246), two potential start codons ATG(20-22) and ATG(108-110), a polyadenylation signal AATAAA(813-818) and selenocysteine insertion sequence (SECIS) motif AUGA(688-691), UGA(729-731) and AAA(703-705). As a selenoprotein, the active-site motifs and GPx family signature motifs LAFPCNQF(101-108) and WNFEK(165-170) were also found. The order of PHGPx mRNA expression levels was: testes >> heart > brain > epididymis > kidney > liver > lung > spleen > muscle. Real-time PCR and immunohistochemistry results revealed similar expression differences in different age testes, with high expression levels during adolescence. Immunofluorescence results suggested that PHGPx mainly expressed in Leydig cells and spermatids in mature goat testes.

Effect of elemental nano-selenium on semen quality, glutathione peroxidase activity, and testis ultrastructure in male Boer goats.

The objective of this experiment is to study the effects of novel elemental nano-selenium in the diet on testicular ultrastructure, semen quality and GSH-Px activity in male goats. Forty-two 2-month-old bucks were offered a total mixed ration which had been supplemented with nano-Se (0.3mg/kg Se) or unsupplemented (the control group only received 0.06mg/kg Se-background), for a period of 12 weeks (from weaning to sexual maturity). Results showed that the testicular Se level, semen glutathione peroxidase and ATPase activity increased significantly in the nano-Se supplementation group compared with control (P<0.05). The semen quality (volume, density, motility and pH) was not affected by added Se in diets, however, the sperm abnormality rate of control bucks was significantly higher than Se supplemented bucks (P<0.05). The testes of 5 goats in each group were examined by transmission electron microscopy (TEM), and showed that in Se-deficient bucks the membrane was damaged, and showed the occurrence of abnormalities in the mitochondria of the midpiece of spermatozoa. In conclusion, selenium deficiency resulted in abnormal spermatozoal mitochondria, and supplementation with nano-Se enhanced the testis Se content, testicular and semen GSH-Px activity, protected the membrane system integrity and the tight arrayment of the midpiece of the mitochondria. Further studies are required to research the novel elemental nano-Se with characterization of bioavailability and toxicity in small ruminants.