Search

Copyright of Veterinarska Stanica is the property of Croatian Veterinary Institute and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Veterinarska Stanica is the property of Croatian Veterinary Institute and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Superovulation is an important step in assisted reproductive technology. Due to its short half-life, follicle stimulating hormone is usually given twice daily to ewes for three to five days, which is both time- and labour-intensive. However, dissolving follicle stimulating hormone in degradable polymers to delay absorbtion has been effective in ruminants. Experiment 1 was performed to compare a split-single follicle stimulating hormone dissolved in hyaluronan (S group; 150 mg follicle stimulating hormone on the first day and 30 mg 48 h later; n = 21) and six decreasing doses of follicle stimulating hormone (M group; 50, 50, 30, 30, 10 and 10 mg; n = 22) at 12-h intervals. Ovarian responses and numbers of recovered ova/embryos did not differ significantly between groups. However, there tended to be more Grade 1 and 2 embryos in S vs M groups (mean ± SEM, 5.1 ± 4.9 vs 2.9 ± 2.9, respectively; P = 0.08). Experiment 2 tested the effectiveness of a simplified split-single follicle stimulating hormone in purebred sheep on a commercial farm. The numbers of recovered good-grade embryos (day 2) were 4.8 ± 5.0 and 4.0 ± 2.5 per donors in Corriedale and Bond sheep breeds, respectively. We conclude that this modified technique for ewe superovulation improved animal welfare, reduced animal handling and labour and yielded results similar to or better than conventional twice-daily follicle stimulating hormone treatments. [ABSTRACT FROM AUTHOR]

Abstract Background The purpose of this study was to apply an arbitrarily primed methylation sensitive polymerase chain reaction (PCR) assay called Amplified Methylation Polymorphism Polymerase Chain Reaction (AMP PCR) to investigate the methylation profiles of somatic and germ cells obtained from Holstein bulls. Methods Genomic DNA was extracted from sperm, leukocytes and fibroblasts obtained from three bulls and digested with a methylation sensitive endonuclease (HpaII). The native genomic and enzyme treated DNA samples were used as templates in an arbitrarily primed-PCR assay with 30 sets of single short oligonucleotide primer. The PCR products were separated on silver stained denaturing polyacrylamide gels. Three types of PCR markers; digestion resistant-, digestion sensitive-, and digestion dependent markers, were analyzed based on the presence/absence polymorphism of the markers between the two templates. Results Approximately 1,000 PCR markers per sample were produced from 27 sets of primer and most of them (>90%) were digestion resistant markers. The highest percentage of digestion resistant markers was found in leukocytic DNA (94.8%) and the lowest in fibroblastic DNA (92.3%, P ≤ 0.05). Spermatozoa contained a higher number of digestion sensitive markers when compared with the others (3.6% vs. 2.2% and 2.6% in leukocytes and fibroblasts respectively, P ≤ 0.05). Conclusions The powerfulness of the AMP PCR assay was the generation of methylation-associated markers without any prior knowledge of the genomic sequence. The data obtained from different primers provided an overview of genome wide DNA methylation content in different cell types. By using this technique, we found that DNA methylation profile is tissue-specific. Male germ cells were hypomethylated at the HpaII locations when compared with somatic cells, while the chromatin of the well-characterized somatic cells was heavily methylated when compared with that of the versatile somatic cells.

Single Layer Centrifugation (SLC) through a low density colloid offers an alternative solution to antibiotic use in boar semen extenders, with lower costs compared to high density colloids. The aim of this study was to explore the reproductive performance of sows when using SLC-prepared semen doses without antibiotics, employing low density Porcicoll to prepare semen doses for artificial insemination in a commercial swine herd in Thailand. Ejaculates were divided into two equal parts to create insemination doses, with each dose containing 3000 × 10 6 sperm/80 ml for intra-uterine insemination in individual sows. The sows were inseminated twice, with the interval between the two inseminations ranging from 8 to 16 h. The CONTROL group consisted of 206 semen doses treated with antibiotics, prepared for insemination in 103 sows, while the SLC group comprised 194 SLC-prepared semen doses without antibiotics for inseminating 97 sows. Fertility and fecundity traits, including non-return rate, conception rate, farrowing rate, and litter traits (i.e., the total number of piglets born per litter, number of piglets born alive per litter, number of stillborn piglets, and number of mummified fetuses), were compared between groups. Furthermore, data on piglet characteristics, including live-born and stillborn piglets (i.e., the prevalence of stillbirth (yes, no), birth weight, crown-rump length, body mass index (BMI), and ponderal index (PI)), were determined. No significant differences in non-return rate (75.7 % vs. 77.3 %), conception rate (73.8 % vs. 73.2 %), and farrowing rate (71.8 % vs. 73.2 %) were observed between the CONTROL and SLC groups, respectively (P > 0.05). Nevertheless, the total number of piglets born per litter in the SLC group was higher than in the CONTROL group (14.6 ± 0.9 vs. 12.3 ± 0.6, respectively, P = 0.049). Interestingly, the prevalence of stillbirth in the SLC group was lower than in the CONTROL group (6.2 % vs. 11.6 %, respectively, P < 0.001). Moreover, the newborn piglets in the SLC group exhibited higher birth weight and BMI compared to those in the CONTROL group (1.36 ± 0.03 vs. 1.26 ± 0.02 kg, P = 0.005, and 18.3 ± 0.3 vs. 17.3 ± 0.2 kg/m 2 , P = 0.003). In conclusion, employing sperm doses after SLC through a low density colloid in artificial insemination within a commercial breeding operation did not have a detrimental impact on either fertility or fecundity traits but showed potential benefits in increasing the total number of piglets born per litter. Moreover, improvements were observed in the birth weight and body indexes of piglets, and the percentage of stillbirths was reduced. Our findings introduce new possibilities for antibiotic alternatives in semen extenders to reduce the risk of antimicrobial resistance in the swine industry. Additionally, they provide compelling reproductive outcomes supporting the integration of SLC-prepared semen doses into artificial insemination practices., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Anti-Müllerian hormone (AMH) serves as an indirect marker for predicting primordial follicles that are representative of ovarian reserve. In this study the possibility of using AMH and age to predict the ovarian reserve in domestic cats. Ovaries and blood were collected from 30 cats undergoing routine ovariohysterectomy. The animals were divided into three age groups: prepubertal (<4 mo, n = 10), adult (1-5 y, n = 10), and senior (>5 y, n = 10). Blood was collected at surgery for serum AMH measurements using the AMH Gen II ELISA kit. The intra-assay coefficient of variation (CV) and inter-assay CV were 3.56 % and 7.68 %, respectively. One side of the ovary was processed to determine AMH localization using immunohistochemistry and for a histological count of follicles, which is the gold standard. The expression of AMH protein was quantified from the contralateral ovary by Western blot analysis. Primordial follicles exhibited the most pronounced inverse relationship with age (rho = -0.779, P < 0.05), followed by a positive association with serum AMH concentration (rho = 0.490, P < 0.05), indicating that both age and AMH are potential markers indicative of primordial follicles. Furthermore, secondary (rho = 0.651, P < 0.05) and small antral follicles (rho = 0.648, P < 0.05) were identified as the major sources of circulating AMH, as indicated by the stronger correlation with serum AMH concentrations compared with primary follicles. However, there was no significant correlation between the expression of AMH protein and other factors, including age, primordial follicles, primary follicles, secondary follicles, small antral follicles, and serum AMH concentration. A model for predicting primordial follicle number using serum AMH concentration (AIC = 672.66, P < 0.05) and age (AIC = 668.93, P < 0.05) was established. In conclusion, both serum AMH concentration and age may serve as comparable markers of ovarian reserve in domestic cats. Moreover, AMH is particularly useful in situations where age information is not available., (Copyright © 2024 Elsevier Inc. All rights reserved.)

The present study aimed to investigate the impact of Clostridium butyricum probiotic feed additive on sow and piglet performances, together with alterations in the lipidomic and metabolomic profiles of sow milk. Sixty-four Landrace × Yorkshire crossbred sows and 794 piglets were included. Sows were divided into two groups; i.e., (i) conventional gestation diet (control; n = 35) and (ii) conventional diet added with 10 g/sow/day of probiotic C. butyricum spores (treatment; n = 29) from one week before the estimated farrowing day until weaning (29.6 ± 4.8 days). The sow and piglet performances and incidence of piglet diarrhea were recorded. Changes in gross chemical composition, fatty acid and non-volatile polar metabolite profiles of sow colostrum, transient milk and mature milk were evaluated. The results showed that relative backfat loss in the treatment group (-2.3%) was significantly lower than in control group (11.6%), especially in primiparous sows ( p = 0.019). The application of C. butyricum probiotics in sows significantly reduced the incidence of diarrhea in piglets ( p < 0.001) but no other effect on piglet performance was found. Lipidomic and metabolomic analyses revealed variations in sow colostrum and milk biomolecular profiles, with indicative compounds significantly altered by feeding with the C. butyricum probiotics. In conclusion, the use of C. butyricum probiotics in sows may improve sow body condition and reduce diarrhea incidence in piglets, with underlying changes in milk composition that warrant further investigation. These findings support the potential of C. butyricum as a beneficial feed additive in swine production.

Premating maternal nutrition is crucial for postweaning follicle growth, thereby influencing piglet birth weight in subsequent litters. The present study investigated the impact of supplementing a carbohydrate-rich premating diet in sows on metabolic hormones, subsequent piglet birth weight and reproductive performance. Sows were distributed into three groups, control (n=42) received standard diets; treatment I (n=41) received the same diets supplemented with 500 g of a carbohydrate-rich premating diet from weaning until insemination; treatment II (n=42) received the same diets supplemented with 500 g of a carbohydrate-rich premating diet from 7 days before weaning until insemination. Blood samples were taken from sows around weaning to measure serum insulin-like growth factor-1 and insulin, and blood glucose after feeding. The study found that sows on a carbohydrate-rich diet (treatment II) had higher postprandial glucose (P<0.05) and insulin levels (P=0.06) than others. This diet did not affect overall reproductive performance, but it did increase piglet birth weight and reduce the number of low-birth weight piglets compared to the control (P<0.001) and treatment I groups (P<0.05). Supplementing a carbohydrate-rich premating diet for 7 days before weaning until insemination enhanced postprandial glucose and insulin concentrations in weaned sows. This dietary intervention led to improved subsequent piglet birth weight and reduced the proportion of low-birth weight piglets.

Objective: This study evaluated the effect of Andrographis paniculata (A. paniculata) supplementation in sow diets before and after farrowing on the sow and piglets' performances during early postpartum period and on sows' backfat and longissimus muscle losses during lactation., Methods: Seventy Landrace×Yorkshire sows and their offspring (1,186 piglets) were distributed into three groups: control (n = 31), treatment-250 (n = 18), and treatment-1000 (n = 21). From 110.2±0.7 days of gestation until farrowing (5.8 days) and throughout the lactation period (25.2 days), sows in the control group were given the conventional lactation diet, while sows in the treatment-250 and treatment-1000 groups received supplements of 250 ppm and 1,000 ppm of A. paniculata, respectively., Results: In sows with parity 3-5, piglets from the treatment-1000 group had higher colostrum intake than the control and treatment-250 groups (p<0.05), but not in sows with parity 6-9. Colostrum immunoglobulin G (IgG) increased in treated sows versus controls for parity 6-9 (p<0.05), but was consistent for parity 3-5. Piglet performance until day 3 postnatal was similar across groups (p>0.05). Treatment-250 sows had higher feed intake post-farrowing than treatment-1000 sows (p<0.05). Longissimus loss was less in both treatment groups than control (p<0.05), but backfat loss was similar across groups (p>0.05). Post-partum complications were consistent across groups (p>0.05). Farrowing duration and piglet birth intervals in sows with parity 6-9 were prolonged in the treatment-1000 group., Conclusion: Supplementing with 1,000 ppm A. paniculata for 5.8 days pre-farrowing and 25.2 days post-farrowing enhanced sow colostrum IgG and piglet colostrum intake, while also reducing longissimus loss in sows. However, for sows of parity 6-9, this supplementation led to prolonged farrowing, increased intervals between piglet births, increased stillbirth, and reduced piglet birth weight. These effects should be considered when using A. paniculata supplementation.

The aim of this study was to explore the factors contributing to colostrum production and the levels of colostrum immunoglobulins (IgG and IgA) in contemporary highly productive sows within a tropical climate. We focused on variables such as parity number, litter size, sow body condition score (BCS), the timing of sample collection following the commencement of farrowing and the use of carbetocin during the birthing process. A total of 100 colostrum samples were collected from a group of 50 Danish Landrace × Yorkshire crossbred sows. These samples were taken at two distinct time intervals: right after farrowing (0 h) and 6 h later. The colostrum samples were classified according to the sows' parity numbers, with 33 samples originating from primiparous sows and 67 from multiparous ones. Additionally, the number of live-born piglets were categorized into three groups: 7-13, 14-17 and ≥ 18 piglets per litter. Moreover, the samples were categorized based on the use of carbetocin during the birthing process, with 34 sows experiencing natural farrowing and 66 sows receiving carbetocin. The sow's BCS was assessed through visual evaluation and palpation. The piglet colostrum consumption and the amount of colostrum produced by the sows were determined. The concentrations of IgG and IgA were determined by using the enzyme-linked immunosorbent assay (ELISA) technique. On average, the colostrum production averaged 5.5 ± 1.7 kg, with IgG and IgA concentrations averaging 54.9 ± 24.6 mg/ml and 7.6 ± 3.5 mg/ml, respectively. Primiparous sows exhibited a significant 25.2% decrease in IgG concentration within 6 h of parturition (P < 0.05), whereas no such decline was observed in multiparous sows. Furthermore, multiparous sows displayed higher colostrum yields (6.2 ± 1.5 kg and 4.3 ± 1.5 kg, respectively, P < 0.001) and IgA concentrations compared to primiparous sows (8.3 ± 3.8 mg/ml and 6.3 ± 2.6 mg/ml, respectively, P = 0.002). Furthermore, a positive correlation was observed between IgA concentrations in colostrum and the sow's BCS at both the 0-h and 6-h post-farrowing time points (r = 0.425, P = 0.002 and r = 0.315, P = 0.031, respectively). The administration of carbetocin did not yield a significant impact on the concentrations of IgG and IgA in the sows' colostrum (P > 0.05). In conclusion, during the initial 6 h after birth, colostrum IgA levels remained stable, whereas there was a noticeable decline in IgG levels, particularly among primiparous sows. The production volume of colostrum and the concentration of IgA in sows within tropical conditions were influenced by both parity number and body condition score., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Anti-Müllerian hormone (AMH) is produced by granulosa cells of the antral follicles. It serves as a promising biomarker for ovarian reserve and responsiveness to ovarian stimulation in humans and domestic animals. This study aimed to validate the AMH Gen II enzyme-linked immunosorbent assay (ELISA) and correlate ovarian structures with serum AMH concentrations after stimulation treatment in clouded leopards (Neofelis nebulosa). Serum samples were collected from 12 women (age 6.21 ± 3.56 years), and serum AMH concentrations were analysed using AMH Gen II ELISA. The animals were divided into two groups based on ovarian structures [preovulatory follicles (>2 mm) and/or corpora hemorrhagica] along with the presence of uterine tonicity visualized laparoscopically around the time of ovulation. Animals that exhibited these reproductive features were identified as the responder group (n = 9, aged 7.59 ± 2.96 years), whereas those lacking the corresponding features were assigned to the nonresponder group (n = 3, aged 2.06 ± 0.53 years). The intra-assay coefficient of variation (CV) and interassay CV was 3.56% and 7.75%, respectively. The linearity of AMH dilution was confirmed (r 2  = .998), and the percentage of recovery ranged from 93% to 115%. The results demonstrated that overall serum AMH concentrations around the time of ovulation were negatively correlated with age (r s  = -.692, p = .013). However, serum AMH concentrations were not correlated with the average number of ovarian structures (r s  = -.535, p = .074). Thus, AMH Gen II ELISA was validated in clouded leopards. Around the time of ovulation, serum AMH decreased with advancing age and ovarian responsiveness cannot be evaluated using serum AMH., (© 2024 Wiley-VCH GmbH.)

This study aimed to determine phosphorus and vitamin B12 supplementation effect in semen extender on the quality and fertility ability of chilled Thai native rooster semen. Eighty-four ejaculates of semen from 26 Thai native roosters (Burmese × Vietnam crossbreed) were included. Semen was collected by applying dorsal-abdominal massage once a week, pooled, diluted to 500 million sperms per dose, and divided into 6 groups. The semen samples used for control group were diluted with modified Beltsville poultry semen extender (BPSE). For the treatment groups 2 to 6: semen samples were diluted with modified BPSE and enriched with phosphorus and vitamin B12 (Octafos Octa Memorial Co., Ltd., Bangkok, Thailand) at concentrations 0.02, 0.04, 0.06, 0.08, and 0.10%. Semen fertility ability was tested in 6 replications by inseminating layer hens. Thirty-six Thai native hens were randomly assigned to 3 groups (control, 0.04, and 0.08%) of 12 hens and were inseminated with a dose of 0.2 mL on collecting day. Sperm motion characteristics (i.e., sperm motility, sperm progressive motility, and sperm kinetic parameters) were measured using a computer-assisted sperm analysis system (SCA, Proiser S.L., Valencia, Spain). Sperm viability, mitochondrial activity, acrosome integrity, plasma membrane integrity, and malondialdehyde (MDA) concentration were also evaluated. The sperm motion characteristics were the highest in the 0.04% supplementation group on all days of collection, especially the VCL and VAP (P < 0.05). The viability, mitochondrial activity, plasma membrane and acrosome integrity of spermatozoa were greater in the 0.04% supplementation group than in the control groups (P < 0.05). The 0.04% supplementation group had the lowest MDA concentration in all days of collection. The 0.04% supplementation group were higher both fertility (66.59 vs. 48.50%: P < 0.05) and hatching rates (58.80 vs. 43.18%: P < 0.05) than in the control group. In conclusion, 0.04% phosphorus and vitamin B12 concentrations supplementation in semen extender improved rooster semen quality and fertility in chilled rooster semen., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Semen cryopreservation is an important technique for preserving the genetic material of numerous species. However, frozen semen is highly susceptible to sperm DNA damage and reduced motility, resulting in decreased fertility. The standard method for cryopreservation and several approaches have not been elucidated. This study aimed to determine the effects of supplementing rooster semen extender with a combination of phosphorus and vitamin B12 on cryopreserved semen quality. Semen was collected weekly via dorso-abdominal massage from 57 Burmese × Vietnam-crossbred Thai native roosters aged 1-3 years. In total, 139 semen samples were collected, pooled, and diluted to 200 million sperm per dose. The pooled sample was divided into six experimental groups: a control group (0.00%) diluted with modified Beltville Poultry Semen Extender (BPSE) and five treatment groups diluted with modified BPSE supplemented with phosphorus and vitamin B12 at concentrations 0.02, 0.04, 0.06, 0.08, and 0.10%, respectively. The semen samples were frozen and evaluated at 0, 15, and 30 min after thawing. Sperm kinematic parameters were determined using a computer-assisted sperm analysis system. Sperm quality was evaluated by measuring sperm viability, mitochondrial activity, acrosome integrity, and plasma membrane integrity. Statistical analyses were performed using a general linear mixed model (MIXED) in SAS. Factors in the statistical model were experimental groups, time after thawing, and interaction between experimental groups and time after thawing. Total and progressive motilities were greater in semen supplemented with 0.04% phosphorus and vitamin B12 compared with those in the control ( p  < 0.05). At 15 min post-thawing, VCL, VAP, and HPA in the 0.04% phosphorus and vitamin B12 supplementation group was greater than that in the control ( p  < 0.05). Phosphorus and vitamin B12 supplementation did not affect sperm kinematics at 0 and 30 min after thawing ( p  > 0.05). All the sperm parameters that were tested for the 0.04% phosphorus and vitamin B12 supplementation group in modified BPSE were the highest at all the timepoints after thawing. Thus, supplementing frozen semen extender with 0.04% phosphorus and vitamin B12 increased sperm motility, sperm kinematic parameters, and sperm quality., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Suwimonteerabutr, Ketkaew, Netiprasert, Khaopong, Osiriphan, Sriamornrat and Nuntapaitoon.)

The present study was carried out to determine the seminal microbiota of boars and their correlation with sperm quality. A total of 17 ejaculates were collected from 17 Duroc boars and were classified according to sperm quality into two groups: low-quality ( n = 8) and high-quality ( n = 9). Each ejaculate was subjected to (i) semen evaluation, (ii) bacterial culture and MALDI-TOF identification, and (iii) 16S rRNA gene sequencing and bioinformatic analyses. No difference in the total bacterial count, alpha diversity, and beta diversity between the high-quality group and the low-quality group was detected ( p > 0.05). While Globicatella sanguinis was negatively correlated with sperm quality ( p < 0.05), Delftia acidovorans was positively correlated with sperm quality ( p < 0.05). Lactobacillales (25.2%; LB) and Enterobacterales (10.3%; EB) were the most dominant bacteria and negatively correlated: EB = 507.3 - 0.5 × LB, R 2 = 0.24, p < 0.001. Moreover, the abundance of Escherichia-shigella was negatively correlated with LB (r = -0.754, p < 0.001) and positively correlated with Proteus (r = 0.533, p < 0.05). Alysiella was positively correlated with Lactobacillus (r = 0.485, p < 0.05), Prevotella (r = 0.622, p < 0.01), and Staphylococcus (r = 0.489, p < 0.05). In conclusion, seminal microbiota is significantly associated with boar semen qualities. The distributions of the most dominant bacterial genera, the differences in the abundance of small subset microbes, and their correlation appear to have far more impact than the overall seminal bacterial content (e.g., total bacterial count, alpha diversity, and beta diversity) on sperm quality.

Objective: Inflammation and pain management in postpartum hyperprolific sows is currently an important animal welfare issue in the swine industry. The present study investigates effects of ketoprofen treatment on the incidence of post-parturient disorders, feed intake, colostrum yield, piglet colostrum intake, colostrum immunoglobulin G (IgG) and piglet mortality rate during the first 3 days of postnatal life., Methods: In total, 61 Danish Landrace×Yorkshire crossbred sows and their offspring (n = 833) were included in the experiment. The sows were randomly distributed into two groups: i) control (n = 31), sows were treated with tolfenamic acid 2 mg per kg for 2 days postpartum; ii) ketoprofen (n = 30), sows were treated with ketoprofen 3 mg per kg for 2 days postpartum. The farrowing process of the sows was monitored for 24 h daily, and data associated with farrowing were collected. Piglet colostrum intake, sow colostrum yield and colostrum IgG were determined., Results: During the first 3 days postpartum, the incidence of sows that had fever did not differ between control and ketoprofen groups (51.6% and 56.7%, respectively, p = 0.692). Piglet colostrum intake did not differ between control and ketoprofen groups (p = 0.736). However, the proportions of piglets that had inadequate colostrum intake were 71.3%, 22.6%, and 5.4% in those with birth weights of <1.0 kg, 1.0 to 1.29 kg, and ≥1.30 kg, respectively (p<0.001). The piglet mortality rate did not differ between control and ketoprofen groups (p = 0.808)., Conclusion: Administration of ketoprofen in postpartum sows for 2 days can control the evidence of post-parturient disorders in sows as effectively as the use of tolfenamic acid. No deleterious effect of ketoprofen was detected on sow colostrum yield, piglet colostrum intake and piglet mortality. Therefore, ketoprofen can be recommended as an alternative anti-inflammatory drug used in postpartum sows.

Background: In tropical environments, boar semen is prepared either from a boar on the same farm as the sow herd or collected in semen collection centers and then transported to other farms. Thus, the semen doses can be used for artificial insemination either immediately or preserved for 2-3 days. The present study investigated the bacteriospermia and its antimicrobial resistance in relation to boar sperm quality during short-term storage in semen extender with or without antibiotics in Thailand. M&M: In total, 20 Duroc ejaculates were collected. Each ejaculate was diluted in Beltsville Thawing Solution extender either with 0.25 g of gentamicin per liter (ANTIBIOTIC) or without gentamicin (NO-ANITIBIOTIC) to create semen doses containing 3,000 × 10 6 sperm/100 mL. These were stored at 17 °C for 4 days. Semen characteristics and total bacterial count (CFU per mL, log 10 ) were measured after collection and during storage., Results: Sperm viability was decreased by 6.4% for every 1.0 log 10 increase in total bacterial count (p = 0.026) and Staphylococcus spp. were the most frequently isolated across ejaculates. Throughout the 4 days of storage, sperm motility, viability and acrosome integrity in the ANTIBIOTIC group were higher than those in the NO-ANTIBIOTIC group (p < 0.05), while the total bacterial count was lower (1.9 ± 0.1 versus 3.9 ± 0.1 log 10 , respectively; p < 0.001). Without antibiotic supplementation, the total numbers of bacteria counted on days 2 and 3 of storage were higher than those determined on days 0 and 1 (p < 0.001). Differences in semen quality were detected on days 2 and 3 between the NO-ANTIBIOTIC and ANTIBIOTIC groups in high-viability semen (p < 0.05). However, no differences in sperm quality between the NO-ANTIBIOTIC and ANTIBIOTIC groups were detected in the low-viability semen on each storage day (p > 0.05). On the last day of preservation, Globicatella sanguinis (57.2%), Delftia acidovorans (18.9%) and Micrococcus spp. (5.9%) remained as the top three most abundant contaminants in the semen with antibiotic., Conclusion: Our findings contribute new insights toward reducing antibiotics as well as rational antibiotic use in the boar AI industry. The growth of bacteria was significantly greater only after 2 days of preservation in the semen without antibiotic. For semen doses diluted from highly viable ejaculates, it is possible to store for 2 days without any antibiotic supplementation. Moreover, bacterial counts increased at the end of storage in the presence of gentamycin, suggesting the loss of bacteriostatic properties of gentamicin to the growth of bacteria during storage., (© 2023. The Author(s).)

Background: Neuro-immune pathways are engaged in antenatal and postpartum depression., Aims: To determine if immune profiles influence the severity of prenatal depression above and beyond the effects of adverse childhood experiences (ACE), premenstrual syndrome (PMS), and current psychological stressors., Methods: Using the Bio-Plex Pro human cytokine 27-plex test kit, we assayed M1 macrophage, T helper (Th)-1, Th-2, Th-17, growth factor, chemokine, and T cell growth immune profiles as well as indicators of the immune inflammatory response system (IRS) and compensatory immunoregulatory system (CIRS) in 120 pregnant females in the early (<16 weeks) and late (>24 weeks) pregnancy. The Edinburgh Postnatal Depression Scale (EPDS) was used to assess severity of antenatal depression., Results: Cluster analyses showed that the combined effects of ACE, relationship dissatisfaction, unwanted pregnancy, PMS, and upregulated M1, Th-1, Th-2, and IRS immune profiles and the ensuing early depressive symptoms shape a stress-immune-depression phenotypic class. Elevated IL-4, IL-6, IL-8, IL-12p70, IL-15, IL-17, and GM-CSF are the cytokines associated with this phenotypic class. All immune profiles (except CIRS) were significantly associated with the early EPDS score, independent of the effects of psychological variables and PMS. There was a shift in immune profiles from early to late pregnancy, with an increase in the IRS/CIRS ratio. The late EPDS score was predicted by the early EPDS score, adverse experiences, and immune profiles, mainly the Th-2 and Th-17 phenotypes., Conclusions: Activated immune phenotypes contribute to early and late perinatal depressive symptoms above and beyond the effects of psychological stressors and PMS.

This study was performed to monitor estrous patterns and, more importantly, changes in anti-Müllerian hormone (AMH) concentrations during the peri-ovulatory period in deslorelin-induced estrous bitches. Healthy anestrous bitches (n = 4) were used. Estrus and ovulation were monitored after deslorelin implantation. Blood samples were collected for analysis of progesterone, estradiol-17ß and AMH concentrations before implantation (day 0) and on days 6, 8, 10, 12, 14, 16, 18, 20 and 22 after implantation. Six days following treatment, all bitches showed estrus signs. Ovulation took place between days 12 and 15. Circulating AMH concentrations varied among bitches from 0.12 to 3.08 ng/mL. However, no significant differences in AMH levels (mean ± SD) were observed between day 0 and days following post-implantation (p > 0.05). There were no significant correlations between AMH and estradiol or AMH and progesterone (p > 0.05). Ultrasonographically, the number of clearly identifiable ovarian follicles was higher before ovulation and the area of ovaries increased after ovulation (p < 0.05). Except for AMH, changes in vaginal cytology, estradiol-17ß and progesterone levels observed in our study were similar to naturally occurring estrus. Large intra- and inter-individual variation in AMH were observed suggesting that AMH is currently not suitable as a canine fertility marker to monitor ovarian response to deslorelin treatment for estrus induction.

Lyophilisation is an alternative method for sperm preservation. The aim of this study was to evaluate the effects of freeze-thawing (F/T) and freeze-drying (F/D) on the quality of epididymal goat sperm. Sperm from each region of the epididymis (caput, corpus and cauda) were collected and evaluated for the expression of phospholipase C zeta (PLC-ζ), protamine 1 (PRM1), transition protein 1 (TNP1) and 2 (TNP2). The effects of F/T and F/D on sperm quality in terms of PLC-ζ expression, chromatin stability (Chromomycin A3; CMA3) and DNA integrity were examined. The fertilising ability after intracytoplasmic sperm injection (ICSI) was also tested. Fresh sperm existed PLC-ζ, PRM1, TNP1 and TNP2, irrespective of the regions of the epididymis. However, different patterns of PLC-ζ expression were found. Although PRM1, TNP1, TNP2 were still expressed after F/T or F/D, only F/T could preserve the presence of PLC-ζ. For fresh sperm, caput epididymal sperm had the lowest evidence of chromatin stability when compared to sperm harvested from other regions of the epididymis. The F/T and F/D further increased the numbers of CMA3-positive sperm (P < 0.001). In all cases, no CMA3 staining was observed in caudal epididymal sperm. The caudal epididymal sperm had significantly greater proportions of sperm with intact DNA compared with caput and corpus epididymal sperm, especially when F/T and F/D were performed. The fertilisation rates of F/D sperm tended to decrease when compared with F/T sperm (4.2 ± 3.2 vs. 13.6 ± 9.0, P = 0.08). It is concluded that the sperm recovered from the caudal epididymis is suitable for freezing and lyophilisation. However, poor fertilisation rates of F/D sperm were coincidently observed, with a deficit demonstration of PLC-ζ., Competing Interests: Declaration of competing interest The authors declare that they have no conflicts of interest., (Copyright © 2022 Elsevier Inc. All rights reserved.)

The variation of gestation length in sows leads to difficulties performing farrowing supervision. The present study was performed to investigate whether oral administration of altrenogest until 112 days of gestation and double administration of PGF2α at 113 days of gestation can synchronise the onset of parturition in sows and minimise deleterious effects on the incidence of stillbirths and colostrum quality. Additionally, the effects of synchronised farrowing on colostrum yield and piglet birth weight, colostrum intake and survival rate of piglets until seven days of postnatal life were also investigated. In total, 193 Landrace x Yorkshire crossbred sows were randomly allocated according to parity number into two groups, i.e. control (n = 95) and treatment (n = 98). The control sows were allowed to farrow naturally. The treatment sows were orally administered 20 mg per day of altrenogest for four days from 109 to 112 days of gestation and were administered PGF2α twice on day 113 of gestation. Individual body weight at birth and 24 h after birth of piglets in all litters were determined in both control (n = 1609) and treatment (n = 1707) groups. Colostrum consumption of all piglets, colostrum yield, colostrum IgG and serum progesterone of sows were determined. On average, the total number of piglets born per litter were 17.0 ± 3.1. The proportion of sows farrowed before 114 days of gestation was higher in the control than the treatment group (8.4% and 2.0%, respectively, P = 0.05) and 92.8% of sows in the treatment group farrowed on day 114 of gestation. The percentage of stillborn piglets per litter did not differ significantly between control and treatment groups (4.5% and 4.6%, respectively). Colostrum yield of sows did not differ between control and treatment groups (5.52 ± 0.13 and 5.28 ± 0.12 kg, respectively, P = 0.174). However, colostrum intake of piglets was lower in the treatment than the control group (354.7 ± 6.6 and 381.2 ± 7.0 g, respectively, P = 0.012). Colostrum IgG was higher in the control than the treatment group (41.2 ± 1.1 and 37.3 mg per ml, P = 0.013). In conclusion, altrenogest treatment from 109 to 112 days and double administrations of PGF2α on day 113 of gestation can control gestation length in sows. No deleterious effects of this protocol on the incidence of stillbirths and sow colostrum yield were detected. However, piglet colostrum intake and colostrum IgG were compromised. Thus, care of newborn piglets in the treatment group should be considered., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Worldwide heat stress (HS) conditions have a negative impact on dairy cow fertility. However, understanding of the effect of heat stress on endometrial functions is still unclear. The present study aimed to investigate the effects of differential heat exposure conditions on the immune response and prostaglandin biosynthesis of bovine endometrium challenged with bacterial lipopolysaccharide (LPS). Cultures of endometrial cells were grown to confluence at 37 °C (control) and 40.4 °C for 24 h after confluence (short-term heat exposure) and 40.4 °C for 8 days from the beginning of the culture (long-term heat exposure), prior to a challenge by 100 ng/mL LPS for 12 h. LPS altered ALOX12 , IL8 , IL1B , S100A8 , PTGES and AKR1B1 expressions, as well as secretory IL8 and PGF2α. Short-term heat exposure decreased S100A8, IL8 and PGF2α compared with the control temperature, while long-term heat exposure decreased S100A8 and PGF2α. In contrast, HSPA5 expression was not altered by heat exposure or LPS. Indeed, the short-term heat treatment was insufficient for accomplishing the responses of the endometrium to LPS treatment for IL8 , S100A8 and PTGES expressions when compared with other temperature conditions. Our findings showed that heat exposure could compromise endometrium immune response and prostaglandin biosynthesis in different ways based on elevated temperature duration, which could reduce subsequent fertility.

Equex STM paste, a water-soluble detergent, exerts the protective effect of egg-yolk during sperm cryopreservation. This study aims to evaluate the post-thaw quality of rhesus monkeys' epididymal spermatozoa in the Tris-citric-glucose egg-yolk extender, supplemented with or without Equex STM paste (0.5%, v/v) (n = 6). Sperm motility, progressive motility, motion characteristics, viability, acrosome integrity and mitochondrial activity were compared immediately post-thaw. Equex STM paste supplementation significantly improved sperm motility (35.0 ± 4.5 vs. 23.7 ± 5.0%), progressive motility (15.4 ± 2.1 vs. 9.8 ± 2.7%) and percentage of sperm with intact acrosome (30.4 ± 4.5 vs. 26.3 ± 4.6%) compared to the controls, respectively. This is the first report applying Equex STM paste for monkey epididymal sperm cryopreservation and is expected to be beneficial as a model for endangered non-human primates., (© 2022 Wiley-VCH GmbH.)

The objective of this study was to find relationships among serum IGF-1, serum testosterone, seminal plasma IGF-1 concentrations and semen parameters in Asian elephants ( Elephas maximus ). A total of 17 ejaculates (one to three ejaculates/bull) were collected from seven captive elephant bulls by performing rectal massage. Before each ejaculation, blood samples were obtained for serum IGF-1 and testosterone assays. Subsequently, the semen characteristics of each ejaculate were evaluated. Mean serum IGF-1 concentration of elephant bulls was estimated as 326.3 ± 114.6 ng/mL (median, 286.2 ng/mL; range, 167.4-542.7 ng/mL). An increase in serum IGF-1 concentration was found to correlate with the percentage of spermatozoa with intact acrosomes. In addition, IGF-1 concentration was positively correlated with testosterone level. However, seminal IGF-1 concentrations could not be detected. In conclusion, our findings suggest that serum IGF-1 concentration is likely a biomarker of normal testicular functions, particularly spermatogenesis in elephants. Moreover, this commercial IGF-1 ELISA is eligible for analyzing serum IGF-1 concentration in Asian elephants.

Objective: Current study was conducted to determine the effect of postpartum prostaglandin F2α (PGF2α) administration on colostrum and milk yield, colostrum immunoglobulin G (IgG) and piglet growth performance., Methods: In total, 36 sows were included in the experiment. The sows were classified into two groups: i) control (n = 11) and ii) PGF2α (n = 25). Sows in the PGF2α group received 10 mg of PGF2α within an hour after farrowing. The body weight of piglets was measured at 0 and 24 h after birth to estimate colostrum consumption. Colostrum was collected at 1 and 24 h after farrowing to determine IgG concentrations. For milk yield study, the remaining sows in the PGF2α group (n = 23) were divided into two subgroups: i) single PGF2α (n = 12) and ii) multiple PGF2α (n = 11). In the multiple PGF2α, the sows received repeated doses of PGF2α at seven and 14 days postpartum. The piglets' body weight was measured at 0, 1, 5, and 20 days of age. The milk yield of the sows was calculated., Results: Colostrum yield of sows averaged 5.62±2.25 kg. Sows treated with PGF2α postpartum had a higher colostrum yield than control (7.01 and 5.12 kg, p<0.05). The concentration of IgG in colostrum at 24 h in the PGF2α group was higher than the control (31.6 and 17.4 g/L, p<0.05). For primiparous sows, milk yield was highest in the sows treated with multiple doses of PGF2α during lactation and lowest in control sows (10.25 and 7.61 kg, p<0.05). Colostrum intake was higher in the treatment than the control groups (+56.7 g, p<0.05). Primiparous sows treated with multiple doses of PGF2α had a higher litter weight than controls (p<0.01)., Conclusion: Postpartum treatment with PGF2α improved colostrum yield and IgG in multiparous sows and increased colostrum intake of piglets. Multiple administration of PGF2α improved the milk yield and increased litter weight of piglets in primiparous sows.

For experiment one, blood samples were obtained from 200 gilts at 90, 120, 150, 180, and 200 days of age. Serum samples from the 30 youngest (166.1 ± 0.7 days) and 30 oldest (198.8 ± 0.6 days) gilts exhibiting estrus by 200 days, and a further 18 gilts that remained anestrus at 200 days, were assayed for serum concentrations of anti-Mullerian hormone (AMH) and estradiol (E2). Gilts younger at puberty had higher ( p < 0.05) AMH levels than those older at puberty, and both groups had higher AMH levels than anestrus gilts ( p < 0.05). Regardless of age, serum E2 was higher ( p < 0.05) in gilts that achieved puberty than in gilts remaining anestrus. At spontaneous pubertal estrus detection, there was no effect of pubertal age on the number of preovulatory ovarian follicles. For experiment two, 152 prepubertal gilts received an intramuscular (IM) injection of 400 IU eCG plus 200 IU hCG and then received fence-line boar contact to detect estrus onset. Serum AMH concentrations were higher ( p < 0.05) in the first 25 gilts to exhibit puberty than the last 28 gilts, with the first gilts also having more preovulatory follicles ( p < 0.0001). Taken together, these data support an association between serum AMH concentrations and degree of physiological maturity and ovarian follicular development in gilts.

Objective: The aim of study was to investigate the effects of season and single layer centrifugation (SLC) before cryopreservation on post-thaw bull sperm quality in Thailand., Methods: Semen was collected from 6 bulls (Bos indicus) in summer, rainy season and winter 2014 through 2016. Semen characteristics, sperm morphology, sperm kinematics, viability, chromatin structure and mitochondrial membrane were evaluated. Meteorological data were available from the local meteorological station., Results: Season had an effect on semen characteristics in the raw ejaculate, with higher proportions of normal spermatozoa and lower abnormalities in winter than in the other two seasons. Sperm kinematics, viability, DNA fragmentation index, and mitochondrial membrane potential were not different between seasons. Sperm samples selected by SLC had greater normal morphology and a lower proportion with bent tails than controls and higher values of progressive motility (PRO), beat cross frequency, linearity, straightness, wobble (WOB), and lower values of slow motility, velocity average path (VAP), velocity curved line, and amplitude of lateral head displacement than controls. In addition, SLCselection had a favorable effect on PRO, VAP, and WOB that differed among seasons., Conclusion: Our results suggested that these bulls were well adapted to their location, with season having an effect on sperm morphology. Moreover, SLC could be used prior to cryopreservation, regardless of season, to enhance normal morphology and kinematics of bull sperm samples without adversely affecting other parameters of sperm quality. However, there was considerable variation among bulls in DNA fragmentation index, mitochondrial membrane potential and sperm viability. In addition, SLC had a positive effect on sperm morphology and sperm kinematics, which could be expected to influence fertility.

During the cryopreservation process, spermatozoa are exposed to hypertonic solutions contributed by the high concentration of cryoprotectant. During addition and removal of cryoprotectant the spermatozoa are subjected to a substantial osmotic stress. Spermatozoa of different species and different stages of maturation may have different susceptibility to osmotic stress depending on the biology of the cell membrane and this will affect their tolerance to the freezing-thawing stress. The aims of this study were to determine the osmotic tolerance limits for motility, membrane integrity and mitochondrial membrane potential of feline epididymal spermatozoa and to study the effect of osmotic stress on the feline spermatozoa of different epididymal regions. Epididymal spermatozoa from three regions (caput, corpus and cauda) were pre-exposed to various osmolalities (75, 300, 600, 900, 1200 mOsm) in a single step for 10min and returned to 300 mOsm afterward. Percentage of motile spermatozoa was measured subjectively and membrane integrity (SYBR-14 positive cells) was evaluated prior to and after exposure to different osmolalities. The mitochondrial membrane potential (JC1) of spermatozoa were evaluated using flow cytometer and compared between epididymal regions (caput, corpus and cauda). All the parameters were compared using a mixed procedure. The percentage of motile epididymal spermatozoa decreased significantly when spermatozoa were exposed to 75 mOsm and 600 mOsm. Epididymal spermatozoa showed signs of damage when pre-exposed to 900 and 1200 mOsm and returned to isotonic condition as significant reduction of membrane integrity and mitochondrial membrane potential were observed (P<0.05). The plasma membrane of spermatozoa from the cauda epididymal region showed higher susceptibility to osmotic stress than the other regions as demonstrated by a significant difference between regions after return to isotonicity from 900 mOsm (P>0.01) and a difference between caput and corpus after return from 1200 mOsm (P<0.05). The corpus and cauda epididymal spermatozoa had higher percentage of spermatozoa with high mitochondrial membrane potential than those from caput when exposed to 75, 300 and 600 mOsm (P<0.05). In conclusion, a single step exposure to hypertonic solution of greater than 600 mOsm prior to return to isotonic condition can cause severe damage to sperm membrane and mitochondrial membrane potential compared to non-returning (exposure to various osmolality but not returned to isotonic condition). Changes in osmolality impacted mostly on sperm motility. Spermatozoa from cauda epididymis were more susceptible to osmotic stress compared to those from corpus and caput indicating that the maturation changes in the sperm membrane during passage through the epididymis increase susceptibility to the osmotic changes that may occur during sperm cryopreservation., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Generally, laparoscopic artificial insemination (LAI) provides a higher success rate than of cervical insemination in goats. However, the sperm distribution after LAI in goats remains unknown, particularly when frozen-thawed semen is used. This study evaluated the distribution of frozen-thawed goat spermatozoa after LAI and compared the effects of sperm numbers and deposition sites (unilateral and bilateral sites) on pregnancy rate. In experiment 1, the frozen-thawed spermatozoa were stained either with CellTracker Green CMFDA (CT-Green) or CellTracker Red CMPTX (CT-Red), and in vitro evaluations of viability and motility were performed. In experiment 2, the labeled spermatozoa were deposited via LAI into the left (CT-Green) and right (CT-Red) uterine horns (n = 4). After ovariohysterectomy (6 hours after insemination), the distributions of green- and red-colored spermatozoa were assessed via tissue section, flushing, and the oviductal contents were also collected. Experiment 3 was designed to test the pregnancy rates in a group of 120 does after LAI using different numbers of spermatozoa (60 and 120 × 10(6) sperm per LAI) and different deposition sites. The results demonstrated that the fluorochromes used in this study did not impair sperm motility or viability. Frozen-thawed goat spermatozoa can migrate transuterinally after LAI, as evidenced by the observations of both CT-Green- and CT-Red-labeled spermatozoa in both uterine horns. Lower numbers of spermatozoa (60 × 10(6)) that are inseminated unilaterally (either ipsilateral or contralateral to the site of ovulation) can efficiently be used for LAI in goats (with a 56.67% pregnancy rate)., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Equine epididymal sperm are known to be severely sensitive to cryopreservation, in terms of sperm quality and pregnancy rate. The objective of this study was to examine the effects of cholesterol loaded cyclodextrins (CLCs) on the quality of stallion epididymal sperm during cryopreservation. In experiment I, sperm were treated with different concentrations of CLCs: (1) 0mg (control), (2) 1.5mg, (3) 3mg, and (4) 6 mg per 120 × 10(6) sperm. The sperm viability and amount of cholesterol were determined at 15, 30 and 45 min after CLC treatment using viability markers (Ethidium homodimer-1 and Calcein AM) and gas chromatography, respectively. In experiment II, CLC treated sperm (1.5mg CLC per 120 × 10(6) sperm) were fixed and stained with filipin to examine the cholesterol distribution. In experiment III, sperm were treated with CLCs at concentrations of 1.5, 3.0, 6.0 mg per 120 × 10(6) sperm for 15 min, then equilibrated with freezing extender at 4°C for 1h prior to cryopreservation. Epididymal sperm without CLC loading (0mg) were used as the control group. The sperm quality was examined at post-equilibration and 10 min, 2h and 4h after freezing and thawing. The cholesterol was successfully loaded into the plasma membrane of stallion epididymal sperm. The amount of cholesterol was increased in a manner of dose and time dependence, and the filipin-sterol complexes were increasingly labeled over the sperm head. CLCs at 1.5mg/120 × 10(6) sperm significantly improved sperm quality during sperm equilibration and cryopreservation compared to other doses of CLCs and non-CLC control. An increasing concentration and incubation time of CLCs was detrimental to sperm quality. It is concluded that cholesterol loading to the sperm plasma membrane via CLCs decreases chilling sensitivity and also improves epididymal sperm cryopreservability., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Background: The purpose of this study was to apply an arbitrarily primed methylation sensitive polymerase chain reaction (PCR) assay called Amplified Methylation Polymorphism Polymerase Chain Reaction (AMP PCR) to investigate the methylation profiles of somatic and germ cells obtained from Holstein bulls., Methods: Genomic DNA was extracted from sperm, leukocytes and fibroblasts obtained from three bulls and digested with a methylation sensitive endonuclease (HpaII). The native genomic and enzyme treated DNA samples were used as templates in an arbitrarily primed-PCR assay with 30 sets of single short oligonucleotide primer. The PCR products were separated on silver stained denaturing polyacrylamide gels. Three types of PCR markers; digestion resistant-, digestion sensitive-, and digestion dependent markers, were analyzed based on the presence/absence polymorphism of the markers between the two templates., Results: Approximately 1,000 PCR markers per sample were produced from 27 sets of primer and most of them (>90%) were digestion resistant markers. The highest percentage of digestion resistant markers was found in leukocytic DNA (94.8%) and the lowest in fibroblastic DNA (92.3%, P < or = 0.05). Spermatozoa contained a higher number of digestion sensitive markers when compared with the others (3.6% vs. 2.2% and 2.6% in leukocytes and fibroblasts respectively, P < or = 0.05)., Conclusions: The powerfulness of the AMP PCR assay was the generation of methylation-associated markers without any prior knowledge of the genomic sequence. The data obtained from different primers provided an overview of genome wide DNA methylation content in different cell types. By using this technique, we found that DNA methylation profile is tissue-specific. Male germ cells were hypomethylated at the HpaII locations when compared with somatic cells, while the chromatin of the well-characterized somatic cells was heavily methylated when compared with that of the versatile somatic cells.

Objective: To evaluate the efficacy of a novel monoclonal antibody (MAb)-based dot-blot ELISA for detection of Leptospira antigens in urine samples of cattle., Sample Population: Blood and urine samples of 45 test cattle from 5 farms in Chonburi province and 20 control cattle from 2 farms in Khon Kaen province in Thailand., Procedure: Blood and urine samples were assayed (microscopic agglutination test and urine antigen test) for Leptospira infection by use of an MAb-based dot-blot ELISA, and results for the ELISA were compared with those for dark-field microscopy (DFM), microbial culture, and a polymerase chain reaction (PCR) assay., Results: All urine samples with positive results for DFM, microbial culture, PCR assay, or > 1 of these tests also had positive results when tested by use of the MAb-based dot-blot ELISA, except for 1 sample that had positive results only for the PCR assay. Detection limits of the dot-blot ELISA were 10(3) leptospires/mL of urine and 9.3 ng of Leptospira homogenate. Comparison revealed that the diagnostic sensitivity, specificity, efficacy (accuracy), positive predictive value, and negative predictive value for the ELISA were in agreement with results for DFM (100%, 72.72%, 80%, 57.14%, and 100%, respectively), microbial culture (100%, 61.54%, 66.62%, 28.57%, and 100%, respectively), and PCR assay (95.45%, 100%, 91.77%, 100%, and 95.83%, respectively)., Conclusions and Clinical Relevance: The MAb-based dot-blot ELISA is suitable as a tool for detecting leptospires in urine samples of cattle.