Sow mortality attributable to pelvic organ prolapse (POP) has increased in the U.S. swine industry and continues to worsen. Two main objectives of this study were, (1) to develop a perineal scoring system that can be correlated with POP risk, and (2) identify POP risk-associated biological factors. To assess POP risk during late gestation, sows (n = 213) were scored using a newly developed perineal scoring (PS) system. Sows scored as PS1 (low), PS2 (moderate), or PS3 (high) based on POP risk. Subsequently, 1.5, 0.8, and 23.1% of sows scored PS1, PS2, or PS3, respectively, experienced POP. To identify biomarkers, serum and vaginal swabs were collected from late gestation sows differing in PS. Using GC–MS, 82 serum metabolite differences between PS1 and PS3 animals (P < 0.05) were identified. Vaginal swabs were utilized for 16S rRNA gene sequencing and differences in vaginal microbiomes between PS1 and PS3 animals were detected on a community level (P < 0.01) along with differences in abundances of 89 operational taxonomic units (P < 0.05). Collectively, these data demonstrate that sows with greater POP risk have differential serum metabolites and vaginal microflora. Additionally, an initial and novel characterization of the sow vaginal microbiome was determined.
Pigs with severe combined immunodeficiency (SCID) are an emerging biomedical animal model. Swine are anatomically and physiologically more similar to humans than mice, making them an invaluable tool for preclinical regenerative medicine and cancer research. One essential step in further developing this model is the immunological humanization of SCID pigs. In this work we have generated T − B − NK − SCID pigs through site directed CRISPR/Cas9 mutagenesis of IL2RG within a naturally occurring DCLRE1C (ARTEMIS) −/− genetic background. We confirmed ART −/− IL2RG −/Y pigs lacked T, B, and NK cells in both peripheral blood and lymphoid tissues. Additionally, we successfully performed a bone marrow transplant on one ART −/− IL2RG −/Y male SCID pig with bone marrow from a complete swine leukocyte antigen (SLA) matched donor without conditioning to reconstitute porcine T and NK cells. Next, we performed in utero injections of cultured human CD34 + selected cord blood cells into the fetal ART −/− IL2RG −/Y SCID pigs. At birth, human CD45 + CD3ε + cells were detected in cord and peripheral blood of in utero injected SCID piglets. Human leukocytes were also detected within the bone marrow, spleen, liver, thymus, and mesenteric lymph nodes of these animals. Taken together, we describe critical steps forwards the development of an immunologically humanized SCID pig model.
Heat stress (HS) negatively affects both human and farm-animal health and undermines efficiency in a variety of economically important agricultural variables, including reproduction. HS impairs the intestinal barrier, allowing for translocation of the resident microflora and endotoxins, such as lipopolysaccharide (LPS), from the gastrointestinal lumen into systemic circulation. While much is known about the cellular function of heat shock proteins (HSPs) in most tissues, the in vivo ovarian HSP response to stressful stimuli remains ill-defined. The purpose of this study was to compare the effects of HS or LPS on ovarian HSP expression in pigs. We hypothesized that ovarian HSPs are responsive to both HS and LPS. Altrenogest (15 mg/d) was administered per os for estrus synchronization (14 d) prior to treatment and three animal paradigms were used: (i) gilts were exposed to cyclical HS (31 ± 1.4 °C) or thermoneutral (TN; 20 ± 0.5 °C) conditions immediately following altrenogest withdrawal for 5 d during follicular development; (ii) gilts were subjected to repeated (4×/d) saline (CON) or LPS (0.1 μg/kg BW) i.v. infusion immediately following altrenogest withdrawal for 5 d; and (iii) gilts were subjected to TN (20 ± 1 °C) or cyclical HS (31 to 35 °C) conditions 2 d post estrus (dpe) until 12 dpe during the luteal phase. While no differences were detected for transcript abundances of the assessed ovarian HSP, the protein abundance of specific HSP was influenced by stressors during the follicular and luteal phases. HS during the follicular phase tended (P < 0.1) to increase ovarian protein abundance of HSP90AA1 and HSPA1A, and increased (P ≤ 0.05) HSF1, HSPD1, and HSPB1 compared with TN controls, while HS decreased HSP90AB1 (P = 0.01). Exposure to LPS increased (P < 0.05) HSP90AA1 and HSPA1A and tended (P < 0.1) to increase HSF1 and HSPB1 compared with CON gilts, while HSP90AB1 and HSPD1 were not affected by LPS. HS during the luteal phase increased (P < 0.05) abundance of HSPB1 in corpora lutea (CL), decreased (P < 0.05) CL HSP90AB1, but did not impact HSF1, HSPD1, HSP90AA1, or HSPA1A abundance. Thus, these data support that HS and LPS similarly regulate expression of specific ovarian HSP, which suggest that HS effects on the ovary are in part mediated by LPS.
During the last decade, sow mortality due to pelvic organ prolapse (POP) has increased. To better understand the biology associated with POP, sows were phenotypically assessed and assigned perineal scores (PS) based on presumed POP risk and categorized as PS1 (low), PS2 (moderate), or PS3 (high). The study objective was to identify changes in sow vaginal microbiota that may be associated with POP. The hypothesis is vaginal microbiota differs between sows with variable risk for POP, and changes in microbiota during late gestation exist between sows with differing risk. Of the 2864 sows scored during gestation week 15, 1.0%, 2.7%, and 23.4% of PS1, PS2, and PS3 sows, respectively, subsequently experienced POP. Vaginal swabs subjected to 16S rRNA gene sequencing revealed differences in community composition (Bray-Curtis; P < 0.05) and individual operational taxonomic unit (OTU) comparisons between vaginal microbiota of PS1 and PS3 sows at gestation week 15. Further, differences (P < 0.05) in community composition and OTUs (Q < 0.05) were observed in PS3 sows that either did or did not subsequently experience POP. Differences in community structure (alpha diversity measurements; P < 0.05), composition (P < 0.05) and OTUs (Q < 0.05) were observed in gestation week 12 sows scored PS1 compared to week 15 sows scored PS1 or PS3, suggesting sow vaginal microbiota shifts during late gestation differently as POP risk changes. Collectively, these data demonstrate sows with greater POP risk have unique vaginal microflora, for which a better understanding could aid in development of mitigation strategies.
Arginine (Arg) is an important amino acid of pig fetal development; however, whether Arg improves postnatal performance is ill-defined. Therefore, the influence of Arg supplementation at different gestational stages on offspring performance was evaluated in a commercial swine herd. Sows (n = 548) were allocated into 4, diet by stage of gestation treatments: Control (n = 143; 0% suppl. Arg), or dietary treatments supplemented with 1% L-Arg (free-base; Ajinomoto Animal Nutrition North America, Inc., Chicago, IL): from 15 to 45 d of gestation (n = 138; Early-Arg); 15 d of gestation to farrowing (n = 139; Full-Arg); and from day 85 of gestation to farrowing (n = 128; Late-Arg). All offspring were individually identified and weighed at birth; at weaning, a subset was selected for evaluation of carcass performance at market. All data were analyzed using birth weight (BiWt) and age as covariates. Wean weights (WW) and prewean (PW) ADG tended to increase (P = 0.06) in progeny from sows supplemented with Arg, as compared to progeny from Control sows. Preplanned contrast comparisons revealed an increased (P = 0.03) BiWt for pigs from sows receiving 1% L-Arg prior to day 45 of gestation (Early-Arg and Full-Arg; 1.38 kg/pig), as compared to pigs from sows not supplemented prior to day 45 of gestation (Control and Late-Arg; 1.34 kg/pig). No difference in BiWt was observed (1.36 kg/pig; P = 0.68) for Arg supplementation after day 85 of gestation (Full-Arg and Late-Arg), as compared to those not receiving Arg supplementation after day 85 (Control and Early-Arg); although WW and PW ADG were greater (P = 0.02), respectively. A 3.6% decrease (P = 0.05) in peak lean accretion ADG occurred when dams received 1% L-Arg prior to day 45 of gestation (Early-Arg and Full-Arg), however, no other significant differences were detected in finishing growth parameters or carcass characteristics (P ≥ 0.1). Pig mortality rates tended (P = 0.07) to decrease in progeny of dams supplemented Arg after day 85 (3.6%) compared to dams not provided additional Arg during late gestation (4.9%). Collectively, these data suggest that Arg provided during late gestation may improve WW and PW ADG, however, finishing performance was not affected. While Arg supplementation provided some moderate production benefits, further investigation is warranted to comprehensively understand the gestational timing and biological role of Arg supplementation during fetal and postnatal development in commercial production systems.
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