Effects of medical plants on alleviating the effects of heat stress on chickens.

Over the past decades, global climate change has led to a significant increase in the average ambient temperature causing heat stress (HS) waves. This increase has resulted in more frequent heat waves during the summer periods. HS can have detrimental effects on poultry, including growth retardation, imbalance in immune/antioxidant pathways, inflammation, intestinal dysfunction, and economic losses in the poultry industry. Therefore, it is crucial to find an effective, safe, applicable, and economically efficient method for reducing these negative influences. Medicinal plants (MPs) contain various bioactive compounds with antioxidant, antimicrobial, anti-inflammatory, and immunomodulatory effects. Due to the biological activities of MPs, it could be used as promising thermotolerance agents in poultry diets during HS conditions. Nutritional supplementation with MPs has been shown to improve growth performance, antioxidant status, immunity, and intestinal health in heat-exposed chickens. As a result, several types of herbs have been supplemented to mitigate the harmful effects of heat stress in chickens. Therefore, several types of herbs have been supplemented to mitigate the harmful effects of heat stress in chickens. This review aims to discuss the negative consequences of HS in poultry and explore the use of different traditional MPs to enhance the health status of chickens.

Identification of Lipids and Cytokines in Plasma and Follicular Fluid before and after Follicle-Stimulating Hormone Stimulation as Potential Markers for Follicular Maturation in Cattle.

The process of follicle maturation leading to ovulation is a key milestone in female fertility. It is known that circulating lipids and cytokines play a role in the follicle's ability to go through follicular maturation and the ovulatory processes. However, the specific mechanisms are not well understood. We posit that dysregulation of granulosa cells influences the ovarian environment, which tries to adapt by changing released lipids and cytokines to achieve follicular maturation. Eleven non-lactating adult females underwent estrus synchronization with two injections of PGF2α 14 days apart. Daily blood samples were collected for 28 days to monitor steroid hormone production after the second injection. To understand the potential impacts of lipids and cytokines during ovulation, a low-dose FSH stimulation (FSHLow) was performed after resynchronization of cows, and daily blood samples were collected for 14 days to monitor steroid hormone production until ovariectomies. The lipidomic analysis demonstrated increased circulating diacylglycerides and triacylglycerides during the mid-luteal phase and after FSHLow treatment. Cholesteryl esters decreased in circulation but increased in follicular fluid (FF) after FSHLow. Increased circulating concentrations of TNFα and reduced CXCL9 were observed in response to FSHLow. Therefore, specific circulating lipids and cytokines may serve as markers of normal follicle maturation.

Impacts of dietary betaine on rectal temperature, laying performance, metabolism, intestinal morphology, and follicular development in heat-exposed laying hens.

This experiment assessed the influences of betaine (BET; 2000 mg/kg) on rectal temperature (Tr), laying performance, metabolism, intestinal morphology, and follicular development in heat-stressed hens. One-hundred and twenty-eight Hisex white hens (42wks) were housed in 4 battery cages (8 pens/cage; 4 hens/pen) and divided into 4 treatments: 1) thermoneutral (TN) environments and a control diet (TNCON), 2) TN and a diet accompanied with BET (TNBET), 3) heat stress (HS) environments and a control diet (HSCON), or 4) HS and a diet accompanied with BET (HSBET). Following acclimation (15d), hens of TNCON and TNBET remained in TN, while HSCON and HSBET hens were subjected to cyclical HS (5d; 16.9-37.5 °C). Cyclical HS increased Tr compared with TN hens (1.6 °C; P < 0.01), but supplemental BET decreased Tr (0.4 °C; P < 0.01). Relative to TN treatments, HS declined egg production, weight, and mass (18, 4.2, and 26%, respectively; P < 0.01), but BET ameliorated the egg production and mass (13.1 and 16.2%, respectively; P < 0.01). Compared with HSCON, feed conversion ratio and survival rate were improved in HSBET hens (12.3 and 6.25%, respectively; P ≥ 0.03). Relative to TN hens, HS elevated glucose and blood urea nitrogen (BUN) levels (15 and 4%, respectively; P ≤ 0.04). Supplemental BET decreased BUN levels (6.6%; P < 0.01) relative to HSCON hens. Furthermore, HS diminished jejunal villus height and villus surface area (∼27 and 35%, respectively; P < 0.01) relative to TN hens but were unaltered by BET supplementation. Relative to TN hens, HS decreased oviduct's weight, ovary's length, and ovarian primordial and primary follicles count (18, 23, 34 and 44%, respectively; P < 0.01) and caused fibrosis in shell gland (3-fold; P = 0.05). Collectively, HS impaired productivity, metabolism, intestinal architecture, and reproductive efficiency. Feeding BET reduced Tr, improved laying performance, and slightly altered metabolism but did not affect intestinal and follicular measurements in heat-stressed hens.

Sheep with ovarian androgen excess have fibrosis and follicular arrest with increased mRNA abundance for steroidogenic enzymes and gonadotropin receptors.

An androgen excess ovarian micro-environment may limit follicle progression in sheep. Two populations of ewes with divergent follicular fluid androstenedione (A4) were identified in a flock in Jordan: High A4; (A4) ≥ 30 ng/mL, (N = 12) or Control A4 (Control); A4 ≤ 15 ng/mL; (N = 12). We hypothesized High A4 ewes would have increased steroidogenic enzyme mRNA abundance, inflammation, and follicular arrest. Messenger RNA abundance for steroidogenic enzymes StAR, CYP17A1, CYP11A1, and HSD3B1 were increased in theca cells while CYP17A1, CYP19A1, and HSD3B1 were increased in granulosa cells in High A4 ewes compared to Control. Gonadotropin receptor mRNA expression for LHCGR was increased in theca and FSHR in granulosa in High A4 ewes. Messenger RNA expression of FOS when reduced, increases expression of CYP17A1 which was observed in High A4 granulosa cells compared to Control. Furthermore, High A4 ewes had greater numbers of primordial follicles (P < 0.001) and fewer developing follicles compared to Control before, and after 7 d of culture, indicating follicular arrest was not alleviated by cortex culture. Increased fibrosis in the ovarian cortex was detected in High A4 ewes relative to Control (P < 0.001) suggesting increased inflammation and altered extracellular matrix deposition. Thus, this High A4 ewes population has similar characteristics to High A4 cows and women with polycystic ovary syndrome suggesting that naturally occurring androgen excess occurs in multiple species and may be a causative factor in follicular arrest and subsequent female sub- or infertility.

Excess androgen (androstenedione; A4) in ewes can result in ovarian follicular arrest and fibrosis contributing to anovulation in sheep. We have identified a naturally occurring ovarian A4 excess in a sheep population with similar characteristics to High A4 cows, both of which are similar to that in women with polycystic ovary syndrome indicating that several mammalian species experience naturally occurring androgen excess resulting in infertility or follicle arrest. Somatic cells, theca and granulosa, surrounding the egg in High A4 ewes had increased expression of steroidogenic enzymes, similar to that seen in High A4 cows, permitting more ovarian cells to manufacture androgens, which may be the cause of androgen excess. Thus, naturally occurring androgen-excess in domestic livestock females can be utilized as models to research the causes of androgen excess and determine the mechanisms that result in follicular arrest and sub- or infertility.

Effects of dietary betaine on body temperature indices, performance, metabolism, and hematological variables of dairy heifer calves during hot summer conditions.

Background and Aim: Heat stress (HS) can negatively impact farm animal productivity and adversely affect animal welfare worldwide, placing a major financial burden on global livestock producers. Dietary betaine (trimethylglycine) has been known to have several biological functions, which may aid in offering beneficial effects on livestock productivity during HS conditions. However, information on the role of dietary betaine in heat-stressed dairy heifer calves is yet to be documented. Therefore, this study aimed to assess the effects of supplementing dietary betaine on body temperature indices, blood metabolites, productive performance, and complete blood count (CBC) (hematological indices) in hyperthermic dairy heifer calves. Materials and Methods: In total, 14 Holstein heifer calves (4.0 ± 0.9 months old) were individually housed and randomly allocated to one of two dietary treatments: (1) a control diet (CON; n = 7) and (2) a control diet complemented with 21 g/d of natural betaine (BET; n = 7) top-dressed once daily. The experiment lasted for 28 d, during which all animals were subjected to natural cyclic HS conditions (26.1-39.2°C; 73.2-84.0 temperature-humidity index). Rectal temperature (RT) and respiration rate (RR) were measured twice daily (0700 and 1500 h), whereas dry matter intake (DMI) was measured once daily (0800 h). In addition, blood samples (collected from the jugular vein) were analyzed for metabolites and CBC on days 7, 14, 21, and 28. Results: Relative to CON, BET supplementation was able to decrease RT on day 23 of the experiment (p = 0.04). Alternatively, RR was similar between the dietary treatments (p = 0.73). Feeding BET did not affect DMI compared with CON during HS conditions (p = 0.48). Furthermore, compared with CON, BET supplementation did not change leukocytes, neutrophils, lymphocytes, and hematocrit levels during HS conditions (p ≥ 0.17). However, a post hoc analysis indicated that hematocrit levels were decreased in BET-fed calves on day 7 of the study compared with CON calves during HS conditions (p = 0.05). Moreover, circulating glucose, albumin, and triglycerides were found to be similar between dietary treatments (p ≥ 0.55). Conclusion: BET supplementation slightly reduced RT and circulating hematocrit but did not affect other metrics in this HS experiment. More research into the effects of different doses of dietary BET on dairy heifer calves is needed.

Naturally occurring androgen excess cows are present in dairy and beef herds and have similar characteristics to women with PCOS.

Beef cows with excess androstenedione (A4; High A4) in follicular fluid (FF) and secreted by the ovarian cortex have been reported from the University of Nebraska-Lincoln physiology herd displaying characteristics reminiscent of polycystic ovary syndrome (PCOS). Thus, we hypothesized that naturally occurring High A4 cows were present in other dairy and beef herds. Fourteen Jordan (Amman, Jordon) dairy heifers and 16 U.S. Meat Animal Research Center beef heifers were classified by FF (High A4: A4 > 40 ng/mL and Control: A4 < 20 ng/mL) and/or cortex culture media (High A4 > 1 ng/mL/d or Control < 1 ng/mL/d). High A4 dairy heifers (n = 6) had greater A4 concentrations (7.6-fold) in FF and (98-fold) greater in ovarian cortex culture media with greater numbers of primordial and fewer later-stage follicles than Controls (n = 8) even after 7 d of culture. Also, the ovarian cortex had greater staining for Picro Sirius red in High A4 dairy heifers compared with Controls indicating increased fibrosis. Thecal cells from High A4 dairy heifers had greater STAR, LHCGR, CYP17A, CD68, and PECAM mRNA expression with increased mRNA abundance of CYP17A1 and CD68 in the ovarian cortex cultures compared with Control dairy heifers. Similarly, cortex culture media from High A4 beef heifers (n = 10) had increased A4 (290-fold; P ≤ 0.001), testosterone (1,427-fold; P ≤ 0.001), and progesterone (9-fold; P ≤ 0.01) compared with Control heifers with increased primordial follicles and decreased later-stage follicles even after 7 d of culture, indicating abnormal follicular development. High A4 ovarian cortex cultures from beef heifers also had increased fibrosis markers and greater expression of PECAM (P = 0.01) with a tendency for increased vascular endothelial cadherin compared with Controls (n = 6). These two trials support our hypothesis that naturally occurring androgen excess cows are present in other dairy and beef herds. The ability to identify these females that have excess A4 ovarian microenvironments may allow for their use in understanding factors causing abnormal follicle development linked to androgen excess and inflammation.

Androgen steroid hormones, normally present in the male, but produced in excess in the female, can result in inflammation and dysfunction of tissues, which, in turn, can lead to ovulatory dysfunction. We have previously identified females with naturally occurring excess androgen in our research herd. In the current paper, we have also identified two other cow populations (one dairy and one beef) that have similar excess androgen production. This suggests that these excess androgen females occur naturally and may be used as models to study androgen excess situations that contribute to subfertility.

VEGFA165 can rescue excess steroid secretion, inflammatory markers, and follicle arrest in the ovarian cortex of High A4 cows†.

A population of cows with excess androstenedione (A4; High A4) in follicular fluid, with follicular arrest, granulosa cell dysfunction, and a 17% reduction in calving rate was previously identified. We hypothesized that excess A4 in the ovarian microenvironment caused the follicular arrest in High A4 cows and that vascular endothelial growth factor A would rescue the High A4 phenotype. In trial 1, prior to culture, High A4 ovarian cortex (n = 9) had greater numbers of early stage follicles (primordial) and fewer later-stage follicles compared to controls (n = 11). Culture for 7 days did not relieve this follicular arrest; instead, High A4 ovarian cortex had increased indicators of inflammation, anti-Mullerian hormone, and A4 secretion compared to controls. In trial 2, we tested if vascular endothelial growth factor A isoforms could rescue the High A4 phenotype. High A4 (n = 5) and control (n = 5) ovarian cortex was cultured with (1) PBS, (2) VEGFA165 (50 ng/mL), (3) VEGFA165B (50 ng/mL), or (4) VEGFA165 + VEGFA165B (50 ng/mL each) for 7 days. Follicular progression increased with VEGFA165 in High A4 cows with greater early primary, primary, and secondary follicles than controls. Similar to trial 1, High A4 ovarian cortex secreted greater concentrations of A4 and other steroids and had greater indicators of inflammation compared to controls. However, VEGFA165 rescued steroidogenesis, oxidative stress, and fibrosis. The VEGFA165 and VEGFA165b both reduced IL-13, INFα, and INFß secretion in High A4 cows to control levels. Thus, VEGFA165 may be a potential therapeutic to restore the ovarian steroidogenic microenvironment and may promote folliculogenesis.

Attainment and maintenance of pubertal cyclicity may predict reproductive longevity in beef heifers†.

We hypothesized the manner that heifers achieve puberty may indicate their future reproductive longevity. Heifers with discontinued or delayed cyclicity during puberty attainment may have irregular reproductive cycles, anovulation, and infertility in their first breeding season contributing to a shorter reproductive lifespan. Therefore, plasma progesterone (P4) was measured from weaning to breeding on 611 heifers born 2012-2017 and four pubertal classifications were identified: (1) Early; P4 ≥ 1 ng/ml < March 12 with continued cyclicity, (2) Typical; P4 ≥ 1 ng/ml ≥ March 12 with continued cyclicity, (3) Start-Stop; P4 ≥ 1 ng/ml but discontinued cyclicity, and (4) Non-Cycling; no P4 ≥ 1 ng/ml. Historical herd records indicated that 25% of heifers achieved puberty prior to March 12th in the 10 years prior to the study. Start-Stop and Non-Cycling yearling heifers were lighter indicating reduced growth and reproductive maturity traits compared with Early/Typical heifers. In addition, Non-Cycling/Start-Stop heifers were less responsive to prostaglandin F2 alpha (PGF2α) to initiate estrous behavior and ovulation to be artificially inseminated. Non-Cycling heifers had fewer reproductive tract score-5 and reduced numbers of calves born in the first 21-days-of-calving during their first breeding season. Within the Start-Stop classification, 50% of heifers reinitiated cyclicity with growth traits and reproductive parameters that were similar to heifers in the Early/Typical classification while those that remained non-cyclic were more similar to heifers in the Non-Cycling group. Thus, heifers with discontinued cyclicity or no cyclicity during puberty attainment had delayed reproductive maturity resulting in subfertility and potentially a shorter reproductive lifespan.

Bovine Ovarian Cortex Tissue Culture.

Follicle development from the primordial to antral stage is a dynamic process within the ovarian cortex, which includes endocrine and paracrine factors from somatic cells and cumulus cell-oocyte communication. Little is known about the ovarian microenvironment and how the cytokines and steroids produced in the surrounding milieu affect follicle progression or arrest. In vitro culture of ovarian cortex enables follicles to develop in a normalized environment that remains supported by adjacent stroma. Our objective was to determine the effect of nutritional Stair-Step diet on the ovarian microenvironment (follicle development, steroid, and cytokine production) through in vitro culture of bovine ovarian cortex. To accomplish this, ovarian cortical pieces were removed from heifers undergoing two different nutritionally developed schemes prior to puberty: Control (traditional nutrition development) and Stair-Step (feeding and restriction during development) that were cut into approximately 0.5-1 mm3 pieces. These pieces were subsequently passed through a series of washes and positioned on a tissue culture insert that is set into a well containing Waymouth's culture medium. Ovarian cortex was cultured for 7 days with daily culture media changes. Histological sectioning was performed to determine follicle stage changes before and after the culture to determine effects of nutrition and impact of culture without additional treatment. Cortex culture medium was pooled over days to measure steroids, steroid metabolites, and cytokines. There were tendencies for increased steroid hormones in ovarian microenvironment that allowed for follicle progression in the Stair-Step versus Control ovarian cortex cultures. The ovarian cortex culture technique allows for a better understanding of the ovarian microenvironment, and how alterations in endocrine secretion may affect follicle progression and growth from both in vivo and in vitro treatments. This culture method may also prove beneficial for testing potential therapeutics that may improve follicle progression in women to promote fertility.

Vascular endothelial growth factor A isoforms modulate follicle development in peripubertal heifers independent of diet through diverse signal transduction pathways.

Follicular progression during peripuberty is affected by diet. Vascular endothelial growth factor A (VEGFA) induces follicle progression in many species; however, there are limited studies to determine if diet may alter the effects of angiogenic VEGFA165-stimulated follicle progression or antiangiogenic VEGFA165b follicle arrest. We hypothesized that diet affects the magnitude of angiogenic and antiangiogenic VEGFA isoform actions on follicular development through diverse signal transduction pathways. To test this hypothesis, beef heifers in our first trial received Stair-Step (restricted and refeeding) or control diets from 8 to 13 months of age. Ovaries were collected to determine follicle stages, measure vascular gene expression and conduct ovarian cortical cultures. Ovarian cortical cultures were treated with phosphate-buffered saline (control), 50 ng/ml VEGFA165, VEGFA165b, or VEGFA165 + VEGFA165b. The Stair-Step heifers had more primordial follicles (P < 0.0001), greater messenger RNA abundance of vascular markers VE-cadherin (P < 0.0001) and NRP-1 (P < 0.0051) than controls at 13 months of age prior to culture. After culture, VEGFA isoforms had similar effects, independent of diet, where VEGFA165 stimulated and VEGFA165b inhibited VEGFA165-stimulated follicle progression from early primary to antral follicle stages. In vitro cultures were treated with VEGFA isoforms and signal transduction array plates were evaluated. VEGFA165 stimulated expression of genes related to cell cycle, cell proliferation, and growth while VEGFA165b inhibited expression of those genes. Thus, VEGFA isoforms can act independently of diet to alter follicle progression or arrest. Furthermore, follicle progression can be stimulated by VEGFA165 and inhibited by VEGFA165b through diverse signal transduction pathways.

Effect of two prostaglandin injections on days 5 and 6 in a timed AI protocol after estrus expression on pregnancy outcomes in dairy cows during cold or hot seasons of the year.

The objective of this study was to test whether prostaglandin (PG) injection on day 30 postpartum (pp) and detection of estrus can affect the efficacy of injecting PG on days 5 and 6 in the timed artificial insemination (TAI) protocol on pregnancy rate in a large dairy herd in hot or cold seasons. Out of 2235 cows, 1998 received an injection of PG at 30 ± 3  d pp and estrus was observed. Cows that displayed estrus during the estrous observation period after PG injection were classified as estrus (E), while those that did not show estrus were classified as nonestrus (NE). Cows in each group were assigned to two treatments: CO-72 (control treatment) (ECO-72 and NECO-72) (day 44 GnRH, day 51 PGF 2 α , day 54 GnRH + TAI) or PG-PG (EPG-PG and NEPG-PG) (day 44 GnRH, day 49 PGF 2 α , day 50 PGF 2 α , day 52 GnRH + TAI). Pregnancy was diagnosed on days 33 and 47 after artificial insemination (AI). The proportion of cows in estrus on the day of TAI was higher ( P ≤ 0.05 ) for cows that received two PG than for cows that received one PG. Pregnancies per AI ( P /AI) on days 33 and 47 for cows inseminated during and after a voluntary waiting period in the NEPG-PG treatment had higher rates than for cows in the EPG-PG, ECO-72 and NECO-72 treatments. Moreover, P /AI were significantly ( P ≤ 0.05 ) affected by parity. Primiparous had higher P /AI (37.0 %) than multiparous cows (31.6 %). Cows inseminated in cold months had higher P /AI and reduced PL (35.6 % and 20.8 %) than cows inseminated in hot months (29.1 % and 30.6 %, respectively). In conclusion, treatments with PG on days 5 and 6 after the first GnRH injection increased P /AI. Estrus detection before the beginning of TAI protocol did not affect fertility. To maximize P /AI cows exhibiting heat at any time during the synchronization protocol should be inseminated.