Non-pregnant Animals Research Articles

Exploration of urinary metabolite dynamicity for early detection of pregnancy in water buffaloes

Early and precise pregnancy diagnosis can reduce the calving interval by minimizing postpartum period. The present study explored the differential urinary metabolites between pregnant and non-pregnant Murrah buffaloes (Bubalus bubalis) during early gestation to identify potential pregnancy detection biomarkers. Urine samples were collected on day 0, 10, 18, 35 and 42 of gestation from the pregnant (n = 6) and on day 0, 10 and 18 post-insemination from the non-pregnant (n = 6) animals. 1H-NMR-based untargeted metabolomics followed by multivariate analysis initially identified twenty-four differentially expressed metabolites, among them 3-Hydroxykynurenine, Anthranilate, Tyrosine and 5-Hydroxytryptophan depicted consistent trends and matched the selection criteria of potential biomarkers. Predictive ability of these individual biomarkers through ROC curve analyses yielded AUC values of 0.6–0.8. Subsequently, a logistic regression model was constructed using the most suitable metabolite combination to improve diagnostic accuracy. The combination of Anthranilate, 3-Hydroxykynurenine, and Tyrosine yielded the best AUC value of 0.804. Aromatic amino acid biosynthesis, Tryptophan metabolism, Phenylalanine and Tyrosine metabolism were identified as potential pathway modulations during early gestation. The identified biomarkers were either precursors or products of these metabolic pathways, thus justifying their relevance. The study facilitates precise non-invassive urinary metabolite-based pen-side early pregnancy diagnostics in buffaloes, eminently before 21 days post-insemination.

Detection of foot-and-mouth disease virus in raw milk in Menofia Governorate and its effect on reproductive hormones and physiochemical properties of milk.

Foot-and-mouth disease (FMD) is an extremely contagious viral disease that affects domestic and wild cloven-hoofed animals. In Egypt, FMD has been enzootic since the 1950s and caused great economic losses in cattle and buffalos over the past few years. This study aimed to detect FMD virus (FMDV) in serum and raw milk samples collected from infected and adjacent cattle and buffalos from different localities in El Menofia Governorate, Egypt. Blood and milk samples were collected from apparently diseased and adjacent 100 cows and 100 buffalos. Serum samples were prepared and used for the detection of FMDV using a non-structural protein enzyme-linked immunosorbent assay, while real-time reverse transcription-polymerase chain reaction (rRT-PCR) was used for the detection of FMDV in milk samples. Reproductive hormones were estimated using radioimmunoassay kits. Milk constituents were determined by Lactoscan. Of the 200 examined serum samples (100 cows and 100 buffalos), 56% and 44% were seropositive for FMDV non-structural protein antibodies in cattle and buffalo, respectively. Real-time reverse transcription-polymerase chain reaction results confirmed that all examined milk samples collected from seropositive animals were positive for FMDV. Estrogen and progesterone levels in the serum of seropositive and seronegative animals were measured, and FMDV was proven to significantly elevate estrogen and reduce progesterone levels in both non-pregnant and pregnant animals during different stages of pregnancy. The effect of the virus on milk composition and somatic cell count (SCC) was also studied, revealing that FMDV infection significantly decreased the level of milk fat, protein, and lactose but did not significantly affected minerals, pH, and conductivity. Moreover, it significantly increased the SCC. Data recorded in this study indicates a widespread occurrence of FMDV in cattle and buffalo all over Menofia Governorate, Egypt. Infected raw milk is of poor quality and, if put for commercial sale, may have health risks for consumers and play a significant role in spreading the virus. Moreover, FMDV may disturb some reproductive hormones, which could adversely affect cattle and buffalo productivity. Therefore, preventive programs and accurate diagnosis are essential for successful disease control.

Trace Elements in Pregnant and Non-pregnant Sheep and Goats and Their Relation to Steroid Hormones

Background: One of the major factors affecting the physiology of pregnancy and reproductive cycle is the trace elements intake and their levels in serum of brood ewes and does. The main objective of this investigation was to study the association between serum concentration of trace elements, ovarian findings, stage of pregnancy and steroid hormones in sheep and goats. Methods: Total of 129 ewes (68 pregnant and 61 non-pregnant) and 54 does (32 pregnant and 22 non-pregnant) were examined clinically and ultrasonographically for pregnancy diagnosis and monitoring the ovarian findings. Pregnant animals were categorized according to the gestational stage into 1 to 2, greater than 2 to 3, greater than 3 to 4 and greater than 4 months. According to the ovarian findings, non-pregnant animals were categorized as having mature corpus luteum (CL), large follicle ( greater than 5 mm) and corpus luteum (CL+F), growing follicles (GF, less than 5.mm) and mature follicle (MF, greater than 5 mm). Animals were bled and serum was harvested and assayed for estrogen (E2), progesterone (P4), manganese (Mn), selenium (Se), iron (Fe) and zinc (Zn) levels. Result: Mn (P=0.01), Fe (P=0.05) and Zn (P=0.001) were lower in pregnant compared with non-pregnant ewes. Fe was significantly higher (P=0.03) in non-pregnant compared with pregnant does at any gestational stage. Ewes with MF had higher Se (P=0.02) and Fe (P=0.001) compared with other ewes. Fe (P=0.001) and Zn (P=0.000) were higher in does having MF compared with other does. A positive association between E2 and serum Fe in non-pregnant ewes (P=0.03) and does (P=0.05) was found. It can be concluded that pregnancy associated with a decrease in serum trace elements in sheep and goats.

Efficiency of Two Protocols of Resynchronization of Estrus and Ovulation in High-Producing Dairy Cows at Peak Lactation

Abstract The reproductive efficiency of the cows was monitored after two resynchronization protocols: Ovsynch (OVS) and Double Ovsynch (DOS). The research initially included 70 HF cows who entered the first synchronization protocol – Presynch. Cows that did not conceive after the first synchronization were divided into two groups and introduced to two resynchronization protocols. In the first group of cows (n=35), the DOS protocol began with the application of GnRH on day 22 after the Presynch TAI (Timed Artificial Insemination), and seven days later pregnancy check-up was done and PGF2α was applied only to non-pregnant cows (n=23), which remained in the study. In the second group of cows, the OVS protocol started on day 32 after Presynch TAI only in non-pregnant animals (n=20). Progesterone (P4) concentration was determined at the time of application of GnRH1, PGF2α and GnRH2 in both groups of cows, and then 30 days after Resynch TAI, ultrasound pregnancy diagnosis was done. A higher percentage of pregnant cows were recorded in the OVS group compared to the DOS group (45% and 35%, respectively). The concentration of P4 in the serum of cows in the DOS group during the first measurement (GnRH1) was significantly higher than the value in cows that did not conceive (p<0.05), while in the third measurement (GnRH2) the average concentration of P4 in conceiving cows was significantly lower (p<0.001) compared to cows that did not coincive. The open days period was significantly longer in pregnant cows that were resynchronized using the DOS protocol compared to cows from the OVS protocol. In conclusion, the OVS protocol of estrus resynchronization in dairy cows proved to be more successful than the DOS protocol. However, considering the advantages the OVS, it is needed to determine which day of the sexual cycle is the best to start resynchronization.

Using Pregnancy-Associated Glycoproteins (PAGs) to Improve Reproductive Management: From Dairy Cows to Other Dairy Livestock.

Simple SummaryPregnancy loss is a major cause of infertility in dairy animals, particularly in cattle, which affects the productivity and profitability of farms. Detecting these unsuccessful pregnancies could offer farmers the opportunity to reduce the economic damage caused by pregnancy loss. The determination of proteins secreted by the placenta and related to the presence of a viable conceptus called pregnancy-associated glycoproteins (PAGs) represents a diagnostic tool to identify pregnant or non-pregnant animals and to predict early pregnancy failures. This review describes the state of the art related to PAGs’ function, pregnancy profile, and use in reproductive management in bovine and other dairy livestock.Pregnancy success represents a major issue for the economic income of cattle breeders. Early detection of pregnant and non-pregnant animals, as well as the prediction of early pregnancy failure, can influence farm management decisions. Several diagnostic tools for pregnancy are currently available. Among these, pregnancy-associated glycoproteins (PAGs) have been shown to be useful for identifying the presence of vital embryos and for pregnancy follow-up monitoring. This review presents an overview of the PAGs’ functions, their pregnancy trends, and their use as a tool to improve reproductive management in bovine and other dairy livestock, such as small ruminants and buffalos.

Use and adequacy of non-pregnancy diagnosis in cow. Which future?

In cattle, early detection of gestation is very important from an economic and management point of view in all types of farming. However, due to the poor efficiency of oestrous detection, it is essential to determine non-pregnant cows as early as possible, in order to minimize the inter-insemination interval, thus de facto, reducing herd open days. Direct and indirect gestation diagnostic methods have been developed with the aim of improving the reproductive performance of the herd. Today, the most accurate method for making an early diagnosis of gestation from 28 to 30 days post-insemination is B-mode ultrasound. In recent years, indirect methods have included techniques that allow non-pregnant cows to be identified with a minimum margin of error, the most widely utilized of which is the colour Doppler. This technique is rapidly becoming established for the diagnosis of non-pregnancy, which allows for the identification of non-pregnant animals earlier compared with the pregnancy diagnosis. Some limitations of this technique in dairy cow have been presented.

Study on Various Luteal Characteristics Using Doppler Ultrasonography for Early Pregnancy Diagnosis in Nili-Ravi Buffaloes.

The objective of current study was to assess the trend in various luteal characteristics viz luteal size (LS), plasma progesterone (P4) concentration, and luteal blood flow (LBF) using color Doppler imaging (CDI) and power Doppler imaging (PDI) modes in pregnant and nonpregnant Nili-Ravi buffaloes. Lactating, cyclic, and healthy Nili-Ravi buffaloes (n = 09) without any reproductive abnormality were selected in present study. Buffaloes were synchronized using Ov-Synch, and fixed-time artificially insemination was performed (day = 0). Pregnancy was diagnosed on 30-day post-AI using B-mode ultrasonography based on presence or absence of embryonic heartbeat. Ovaries of all animals were scanned from day 5 till 21 post-AI using both B-mode and Doppler ultrasonography to measure LS and LBF. After each ovarian ultrasound examination, blood samples were collected via jugular venipuncture to determine plasma P4 concentration. According to results, LBF using CDI and PDI was significantly higher (P ≤ 0.05) in pregnant buffaloes on days 13 and 15 post-AI, respectively. The mean LS and plasma P4 concentration did not differ (P ≥ 0.05) between pregnant and nonpregnant animals until day 15 post-AI. However, a significant difference (P ≤ 0.05) was noticed for both on day 17 and onwards. It is concluded that LBF is a more sensitive luteal character as compared to LS and P4 for earlier pregnancy diagnosis in Nili-Ravi buffaloes when ascertained through CDI.

Histological and hemodynamic characterization of corpus luteum throughout the luteal phase in pregnant and non-pregnant buffalos in relation to nitric oxide levels based on its anatomical determination.

This study aims to compare the complete growth and development of corpus luteum (CL) in domestic buffalos from day 5 until day 40 after ovulation either in pregnant or non-pregnant animals and whether luteal vascularity (LV) with progesterone (P4) and nitric oxide (NO) could determine luteal functionality or not. Pluriparous buffalos (Bubalus bubalis) were categorized as pregnant (n = 6) or non-pregnant (n = 9) after pregnancy check at day 25. Animals were subjected to ultrasound analysis to determine the CL area (cm2) and LV. Blood sampling was performed following the Doppler examination. Ovarian tissue samples from non-pregnant buffalo genitalia (n = 18) and early pregnant buffalo genitalia (n = 3) were collected from great abattoirs. Luteal Doppler indices were lower in the pregnant group, while peak systolic velocity (PSV) was increased (p < 0.05) in the same pregnant females. Both P4 and NOMs were elevated (p < 0.05) in the pregnant group. There was a positive correlation (p < 0.01) between P4 and CL PSV. Based on our macroscopical examination, the CL of non-pregnant buffalos was classified into four stages. Histologically, stage I showed that CL was covered by a highly vascularized connective tissue (CT) capsule. It consisted of small and large lutein cells, whereas stage II was similar to stage I except for the presence of numerous fibroblast cells and vacuolated cells. Stage III was characterized by increasing the number of collagen fibers and the thickness of the blood vessels. Stage IV revealed thickening of the CT capsule and septae, regressed capillaries and arterioles, in addition to shrunken degenerated lutein cells. CL of pregnant buffalos revealed the same structure as CL at stage II. CL area was increased in the pregnant group. The collective data suggested that evaluation of the luteal artery could be extremely helpful to determine the potential benefits of colored and pulsed Doppler in CL vascularization assessment in both luteal and early pregnancy phases.

Ablation of PI3K-p110alpha Impairs Maternal Metabolic Adaptations to Pregnancy.

Pregnancy requires adaptations in maternal metabolism to support fetal growth. The phosphoinositol-3-kinase (PI3K) signalling pathway controls multiple biological processes and defects in this pathway are linked to metabolic disorders including insulin resistance and glucose intolerance in non-pregnant animals. However, relatively little is known about the contribution of PI3K signalling to the maternal metabolic adaptations during pregnancy. Using mice with partial inactivation of the PI3K isoform, p110α (due to a heterozygous dominant negative mutation; Pik3ca-D933A), the effects of impaired PI3K-p110α signalling on glucose and insulin handling were examined in the pregnant and non-pregnant states and related to the morphological, molecular, and mitochondrial changes in key metabolic organs. The results show that non-pregnant mice lacking PI3K-p110α are glucose intolerant but exhibit compensatory increases in pancreatic glucose-stimulated insulin release and adipose tissue mitochondrial respiratory capacity and fatty acid oxidation. However, in pregnancy, mutant mice failed to show the normal increment in glucose intolerance and pancreatic β-cell mass observed in wild-type pregnant dams and exhibited further enhanced adipose tissue mitochondrial respiratory capacity. These maladaptations in pregnant mutant mice were associated with fetal growth restriction. Hence, PI3K-p110α is a key regulator of metabolic adaptations that support fetal growth during normal pregnancy.

Longitudinal patterns in progesterone metabolites in pregnant and non-pregnant Steller sea lions (Eumetopias jubatus)

Similar to the several pinniped and a few terrestrial carnivore species, the Steller sea lion has a seasonal synchronized mating scheme enabled by a female reproductive cycle that includes embryonic diapause, delayed implantation, and pseudopregnancy (a state in which the corpus luteum produces progesterone for approximately as long as in pregnant females). Due to this, circulating systemic progesterone concentrations cannot be used to differentiate pregnant and nonpregnant females during early gestation. With the use of advanced measurement technologies such as liquid chromatography tandem mass spectrometry (LC-MS/MS) additional steroid hormones are measurable which can provide additional information on the endocrine pathways throughout gestation. Our objectives were to further characterize endocrine patterns in female Steller sea lion pregnancy by 1) quantifying longitudinal profiles of hormone metabolites in pregnant and non-pregnant female sera, and 2) evaluating hormone profiles to identify pregnant animals within the early stage of gestation. Three gestation stages were delineated based on what is believed to be the period of implantation (September-October): EARLY (August– November), MID (December-February), and LATE (March to May). Five steroids, Progesterone (P4), 5α-dihydroprogesterone (DHP), 17αOH-progesterone (17OHP), 20αOH-progesterone (20OHP), and androstenedione (A4), were detected in both pregnant and non-pregnant animals. A significant difference in P4 concentrations was measured between EARLY and MID gestation (p ≤ 0.01) in both pregnant and non-pregnant animals. During MID gestation there was a significant difference (p ≤ 0.05) between pregnant and non-pregnant animals in all pregnanes measured. Significant patterns of correlation between P4 and 17OHP and between P4 and DHP were detected during EARLY and MID gestation in non-pregnant animals. While those significant correlations also exist in EARLY pregnant animals, this pattern was lost by MID gestation. This loss of correlation suggests a potential shift in progesterone metabolism from ovarian to alternative tissue (e.g. fetal gonads or adrenal glands) by MID gestation in Steller sea lions. We were unable to identifying a steroid hormone biomarker capable of differentiating pseudopregnancy from pregnant animals and conclude that such a biomarker likely falls outside of the traditional progesterone metabolic pathway.

Evaluation of bovine visual ELISA test for detection of pregnancy-associated glycoproteins in early pregnancy diagnosis in goats

Pregnancy diagnosis is the primary factor for improving reproductive performance and high economic efficiency in herds. Early pregnancy diagnosis, on the other hand, allows livestock to optimise production with greater reproductive performance and correct it with timely intervention. In the present study, it was aimed to investigate the effectiveness and reliability of the commercial rapid visual pregnancy test kit prepared for the detection of pregnancy-associated glycoproteins from blood serum in early pregnancy diagnosis in goats. Healthy and fertile, 3–5 years old 150 Damascus goats were used in the study. Oestrous synchronisation was applied to the goats between the anoestrous period and the onset of the transition period. Goats in oestrous were bred naturally. Blood samples were taken from the goats on the 28th day after their breeding. Progesterone measurement and rapid visual pregnancy test were performed from the blood samples taken. For the diagnosis of pregnancy, transabdominal ultrasonography was applied to the goats 50 days after the breeding. Ultrasonographic examination results were used as reference value. Sensitivity, specificity, positive predictive value, and negative predictive value ratios of serum progesterone measurement and visual PAG-ELISA kit were determined as 88.23%, 91.46%, 89.55%, 90.36% and 94.12%, 80.49%, 80%, 94.29%, respectively. In the study, it was ascertained that progesterone concentration results were in good agreement with ultrasonography results (kappa=0.798), and visual PAG-ELISA results were in good agreement with ultrasonography results (kappa = 0.735). Moreover, the concordance of progesterone concentration results with visual PAG-ELISA results was also found to be excellent (kappa = 0.828). The ability to accurately diagnose serum progesterone measurement and visual PAG-ELISA kit was found to be 90% and 86.67%, respectively. In conclusion, it can be accepted that the visual PAG-ELISA test in goats is useful in distinguishing pregnant from non-pregnant animals in the early stages of pregnancy. It can be said that in practice it can be used alone or in combination with other methods.

Oxytocin, Prostaglandin F2α, and Prostaglandin E2 Induced Contractions in Isolated Uterine Smooth Muscle Strips from Non‐pregnant, Late Pregnant, and Postpartum Rats

Objective/IntroductionNumerous studies have investigated uterotonic agents (e.g., oxytocin) in uterine smooth muscle (USM) from non‐pregnant (NP) and late‐pregnant (LP) women and/or animal models of pregnancy. Surprisingly, few studies have investigated USM from the post‐partum period to determine whether USM contractility returns to its pre‐pregnant state. We designed a study in rodents to investigate whether USM contractility to oxytocin (OXY), prostaglandin F2α (PGF2α), and prostaglandin E2 (PGE2) returned to pre‐pregnant values within 30 days postpartum.MethodsStudies were performed in NP (n=12), LP (21d gestation, n=4), 3 days post‐partum (PP3, n=2), and 30 days post‐partum (PP30, n=4) Sprague Dawley rats. Uterine horns were cleaned of connective tissue and cut along the longitudinal axis allowing isolation of USM (width: 2mm, length: 4±1mm) oriented in the longitudinal muscle (LM) or circular muscle (CM) direction. CM or LM strips were mounted in tissue baths containing Krebs and equilibrated @ 37°C for 60 min prior to starting one of the two following protocols.Protocol 1 identified the optimal resting tension (in grams: 0.25, 0.50, 1.0, and 1.5) to measure spontaneous contraction (SCA), K+‐induced contraction (60mM K+), and cumulative OXY‐induced contraction (10‐11 to 10‐7) in CM and LM of NP rats (N=4‐6).Protocol 2 was designed assess SCA, K+‐induced contraction, cumulative concentration‐response relationships to OXY (10‐11 to 10‐7), PGF2α (10‐8 to 10‐4), or PGE2 (10‐8 to 10‐4) in CM and LM, of NP, LP, PP3, and PP30 rats (N=2‐6).Statistical comparisons were made using either a t‐test or two‐way ANOVA, and considered statistically different at P&lt;0.05.ResultsProtocol 1‐ LM SCA was greater than CM SCA at all resting tensions. K+‐induced constrictions in LM and CM were similar at each resting tension. OXY‐induced contraction in LM was generally greater than CM (P&lt;0.05 at 0.25g, 0.5g, and 1.5g tension, comparable at 0.5, 1.0, and 1.5g tension).Protocol 2 ‐ SCA in NP, LM was greater than NP, CM (P&lt;0.05). There was no difference in SCA in CM vs LM in USM from LP, PP3, and PP30 animals. K+‐induced constrictions in CM and LM were similar in USM from NP, LP, PP3 and PP30 animals. OXY‐induced contractions were greater in LM than CM for all pregnancy states (P&lt;0.05). PGE2 contractions were greater in LM vs CM in USM from NP rats (P&lt;0.05), but were similar in USM from LP, PP3, and PP30 animals. In contrast, PGF2α‐contractions were similar in LM and CM for all pregnancy states.Summary/ConclusionOur preliminary data suggests that uterotonic agents be assessed at either 0.5g or 1.0g resting tension. In addition, uterotonic agents exhibit significantly greater contraction in longitudinal vs circular orientation. Finally, contractility to uterotonic agents returns to pre‐pregnant values by PP30.

Effect of Ovarian Status on Reproductive Performance in Dairy Heifers Inseminated with Sexed Semen

Background: The intensive reproductive management in the dairy farms requires inclusion of a large group of replacement heifers in the breeding program for a shorter period. In this aspect, a creation of effective estrus synchronization protocols with timed artificial insemination (TAI) by sexed semen and optimization of the current ones have a crucial role for obtaining high pregnancy rate. These protocols are beneficial, because they led to reduced interval to first AI, lack of need for estrus detection, and allow obtaining a large group of female calves for on time. Because of limited fertilizing potential of sexed spermatozoa this type of semen is applied mainly for heifers, as fertility is higher compared to lactating cows. The main objective of the present study was to investigate the effect of ovarian status on the reproductive performance in dairy heifers subjected to estrus synchronization and timed artificial insemination with sexed semen. Materials, Methods &amp; Results: Forty-eight Holstein healthy heifers separated in 2 groups were subjected to PGF2α-GnRH pre-treatment and Ovsynch or PRID-5-day estrus synchronization protocol and timed artificial insemination (TAI) with sex-sorted semen, staring 6 days after end of the hormonal pre-treatment. The ovarian status (presence of follicles with or not a corpus luteum) of the heifers at the begin of the treatment, on day of TAI and pregnancy rate in different groups were determined and compared. Additionally, the ovarian status at the begin of the treatment, ovarian status and size of preovulatory follicle (PF) on day of TAI and total values for both groups according to reproductive performance (pregnant or non-pregnant) were also analyzed. On day of TAI the animals with PF and a lack of corpus luteum (CL) in both groups were more that those with PF and CL (39.3% and 30% vs. 60.7 % and 70%), with significant (P &lt; 0.05) difference in PRID-5-day group. The pregnancy rate tended to be higher in PRID-5 day than Ovsynch treatment (65% vs. 35.7%). A higher percentage (100% and 67.9%) of the pregnant animals in both treatments had not CL on day of TAI, and the size of the PF (1.58 ± 0.12 cm and 1.64 ± 0.13 cm) was increased (P &lt; 0.05). Similar effects of the ovarian status on reproductive performance were obtained after a comparison of the total values between pregnant and non-pregnant animals. The percentage of heifers with observed PF without CL on day of TAI was significantly greater (P &lt; 0.05) in pregnant compared to non-pregnant group (91.3% vs. 40%). The opposite dependence was determined for the parameter presence of follicles and corpus luteum (8.7% vs. 60%; P &lt; 0.05). Furthermore, the size of the PF measured immediately before TAI was increased in animals became pregnant (1.60 ± 0.12 cm vs. 1.34 ± 0.17 cm; P &lt; 0.05). Discussion: The analysis of the obtained results showed that the ovarian status on day of TAI affects the reproductive performance in dairy heifers subjected to estrus synchronization and timed artificial insemination with sexed semen. Hormonal pre-treatment with onset of PRID-5-day protocol 6 days later and TAI with sex-sorted semen ensure acceptable pregnancy rate. The greater preovulatory follicle and a lack of corpus luteum before insemination provide significantly (P &lt; 0.05) more pregnant animals, compared to the cases when CL is presented. Ultrasound determination of the ovarian function before insemination can be used in selection of heifers for TAI with sex-sorted semen. Keywords: heifers; estrus synchronization; sexed semen, pregnancy.

Maternal-placental-fetal drug metabolism is altered by late gestation undernutrition in the pregnant ewe

BackgroundMaternal undernutrition during pregnancy disrupts both fetal growth and development with perturbations to certain physiological processes within the maternal-fetal-placental unit, including metabolic function. However, it is unknown if hypoglycemia during pregnancy alters maternal-fetal-placental drug metabolism as mediated by cytochrome P450 (CYP) enzymes. Despite this, hypoglycemia reduces CYP enzyme activity in non-pregnant animals. We therefore hypothesised that in a sheep model of hypoglycemia induced by late gestation undernutrition (LGUN), maternal-fetal-placental CYP activity would be reduced, and that fetal glucose infusion (LGUN+G) would rescue reduced CYP activity. MethodsAt 115d gestation (term, 150d), ewes were allocated to control (100% metabolic energy requirement (MER); n = 11), LGUN (50% MER; n = 7) or LGUN+G (50% MER + fetal glucose infusion; n = 6) and maintained on their diets until post-mortem. Maternal-fetal-placental CYP activity assays were performed at 131-133d gestation. Microsomes were isolated from placenta and fetal liver collected at 139-142d gestation and incubated with CYP-specific probe drugs. Metabolite concentrations were measured using Liquid Chromatography – tandem mass spectrometry (LC-MS/MS). ResultsCYP2C19 and CYP3A were undetectable in placenta or fetal liver, and CYP1A2 was undetectable in the fetal liver. Placental-specific CYP1A2 and CYP2D6 activity and hepatic-specific CYP2D6 activity were unaffected by LGUN. Maternal-fetal-placental CYP1A2 activity was reduced in response to LGUN in the maternal compartment only. ConclusionsReduced maternal-fetal-placental CYP1A2 activity, but not placental-specific CYP1A2 activity, may lead to the developing fetus being exposed to increased concentrations of CYP1A2-specific substrates and suggests further consideration of drug dosing is required in instances of late gestation maternal undernutrition.

CHARACTERISTICS OF SKIN DIELECTRIC PROPERTIES IN PREGNANCY (EXPERIMENTAL STUDY

The purpose of our study was to evaluate the dielectric characteristics of the skin that occur during pregnancy in an experiment. The study included 60 Wistar rats, 30 of which were pregnant (in the 3rd trimester; parturition took place 2–3 days after the study). Once, on a pre-epilated area of the skin of the back, the dielectric properties were studied using the method of near-field resonant microwave sensing. The diagnostic software and hardware complex for microwave sensing allows calculating the level of dielectric permittivity of the medium and conductivity based on the shift of the resonant frequency. Based on the conducted studies, it was found that the integumentary tissues of pregnant rats have a significantly higher level of dielectric permittivity and conductivity compared to non-pregnant animals. Such shifts are caused by the formation of edematous syndrome associated with pregnancy, and predispose to the development of postpartum skin changes.

An Investigation of Ovarian and Adrenal Hormone Activity in Post-Ovulatory Cheetahs (Acinonyx jubatus).

Simple SummaryMore than three decades of scientific study have been unable to determine the etiology of reproductive issues in cheetahs under human care. The reproduction of cheetahs in zoological facilities has never been self-sustaining, and the differences between females who establish pregnancy and those that do not remain poorly understood. The objective of this study was to examine and compare ovarian and adrenal hormones post-ovulation in pregnant and non-pregnant animals to better understand female physiology after natural breeding or artificial insemination, and determine what may be contributing to the frequent lack of success. The authors also sought to validate a urinary progestagen assay to assist with pregnancy detection. Although fecal glucocorticoid metabolites among pregnant and non-pregnant groups were not different, samples from the third trimester in pregnant animals were higher than at any other time. Additionally, glucocorticoids were higher, and estrogens tended to be lower in samples from pregnant females that gave birth to singletons, than those that had multi-cub litters. As a supplementary benefit, this is the first time urinary progestagens have been measured and have been able to distinguish pregnant and non-pregnant cheetahs. The results provide first-time insights into ovarian and adrenal hormonal events surrounding ovulation and pregnancy detection in cheetahs.Cheetahs have been the subject of reproductive study for over 35 years, yet steroid hormone activity remains poorly described after ovulation. Our objective was to examine and compare fecal progestagen (fPM), estrogen (fEM), and glucocorticoid (fGM) metabolite concentrations post-ovulation in pregnant and non-pregnant animals to better understand female physiology (1) during successful pregnancy, (2) surrounding frequent non-pregnant luteal phases, and (3) after artificial insemination (AI) to improve the low success rate. Secondarily, the authors also validated a urinary progestagen metabolite assay, allowing pregnancy detection with minimal sample collection. Fecal samples were collected from 12 females for ≥2 weeks prior to breeding/hormone injection (the PRE period) through 92 days post-breeding/injection. Samples were assessed for hormone concentrations using established enzyme immunoassays. Urine samples were collected for 13 weeks from 6 females after natural breeding or AI. There were no differences among groups in fGM, but in pregnant females, concentrations were higher (p < 0.01) in the last trimester than any other time. For pregnant females that gave birth to singletons, fGM was higher (p = 0.0205), but fEM tended to be lower (p = 0.0626) than those with multi-cub litters. Our results provide insight into the physiological events surrounding natural and artificially stimulated luteal activity in the cheetah.

Expression Level of the mip, pmp18D, and ompA Genes in Chlamydia abortus Isolated from Aborted Ewes

In this manuscript, we report the proteins macrophage infectivity potentiator (mip, CAB080), major outer membrane protein (momp, CAB048), and polymorphic outer membrane protein (pmp18D, CAB776) that are expressed in different times of pregnancy in mice infected with Chlamydia abortus. Enzootic abortion of ewes (EAE) by C. abortus, an obligate intracellular pathogen, is a critical zoonotic disease-causing significant economic loss to livestock farming globally. This study was carried out for the detection and characterization of macrophage infectivity potentiator (mip, CAB080), major outer membrane protein (momp, CAB048), and polymorphic outer membrane protein (pmp18D, CAB776) using RT-qPCR. These proteins are believed to be expressed as virulence factors in C. abortus isolated from aborted ewes. BALB/c mice (pregnant and nonpregnant) were used as an animal model to be injected intraperitoneally with C. abortus culture in Vero cells since the endometrial lymphoid tissues of these animals resembles that of ewes. Also, the short duration of pregnancy in mice makes them a suitable animal model for obstetric studies. Tissue samples were taken from the mice after 10, 15, and 20 days of pregnancy to compare the expression of the genes mip, pmp18D, and ompA. Transcription level was quantified using RT-qPCR, the GAPDH transcription quantification, as a normalization signal. Abortion occurred in pregnant mice, and apparent differences between the transcriptional levels of the mip, pmp18D, and ompA genes in the samples taken during different time intervals of pregnancy were not observed (p > 0.05). The result indicated that the three bacterial genes, mip, pmp18D, and ompA, play a role as virulence factors in abortion and are differentially expressed in pregnant and nonpregnant animals. Inactivation of the genes is suggested to confirm the hypothesis.

Estimation of co-variance components and genetic parameters of fertility and production traits in crossbred cattle of Kerala

Estimation of co-variance components and genetic parameters of fertility and production traits will help to find out the relative importance of genetic and environmental components of each trait and to develop a genetic evaluation system for overall improvement in performances of crossbred cattle of Kerala. In the present study, major fertility trait considered was daughters pregnancy rate (DPR), measures the percentage of non-pregnant animals that become pregnant during each oestrous cycle. Data pertaining to 1180 crossbred cattle sired by 208 Frieswal bulls, spread over a period of 16 years from 2003 to 2019, maintained at different farms of Kerala Veterinary and Animal Sciences University and field centres of ICAR- Filed Progeny Testing Scheme were analysed in the study. Estimates of covariance components and genetic parameters were obtained using restricted maximum likelihood (REML) approach using average information (AI) algorithm. It was observed that DPR had low heritability (0.092 ± 0.03), compared to 305 days milk yield (MY) (0.170 ± 0.094) and fat percent (FP) (0.173 ± 0.072). Phenotypic (rp), genetic (rg) and residual (re) correlation indicated unfavourable association of fertility with production traits. The estimates of variance and (co)variance components were computed by multivariate animal model. The results indicated that DPR was having lower direct additive (σ2a) 0.046 and environmental variance (σ2c) 0.063 compared to other traits. Highest additive genetic variance (σ2a) 27035.8 was obtained for MY. The study estimates the magnitude of correlations and covariances of fertility and production traits. Since DPR had lower additive genetic variance and negative association with milk production traits in cattle, it would be included as an indirect measure in the evaluation and breeding programs of crossbred cattle of Kerala.

Fecal Cortisol Dynamics of Captive Giraffes in Zoos in Northern Japan

Animals in zoos are kept under environmental conditions differing from those of their original habitat. Recently, zoos have been attempting to reduce stress by introducing environmental enrichment. Adrenal-derived glucocorticoids are common stress indicators found in blood, saliva, urine, and feces. Fecal cortisol levels have been studied as a stress marker in captive giraffes, but the data are insufficient. In this study, we examined fecal cortisol in captive giraffes in northern Japan to increase basic data on fecal cortisol as an indicator of environmental enrichment. For one month, we collected fecal samples from 14 giraffes [Masai giraffes (Giraffa camelopardalis tippelskirchi) and reticulated giraffes (Giraffa camelopardalis reticulata)] from eight zoos. The fecal samples were stored at –20 °C until fecal cortisol levels were measured using a cortisol EIA kit after drying, grinding, and ethanol extraction. Cortisol levels were compared between males and females, pregnant and nonpregnant animals, by age, and by differences in temperature of the rearing environment. The fecal cortisol levels ranged from 37~10346 pg/g. Among females, pregnant individuals had the highest values. Among nonpregnant females, a female that was harassed by a male had higher fecal cortisol levels than the others. This may be due to chronic psychological distress. In males, values varied greatly among individuals. This study increases the volume of giraffe fecal cortisol data, which will contribute to establishing basic giraffe fecal cortisol values.

Epidemiology of Gastrointestinal Nematodes in Small Ruminants in Gamo Zone, Southern Ethiopia, with a Special Reference to Field Survey on Parasite Management and Risk Factor Analysis

Gastrointestinal nematodes are a major constraint for small ruminant production. A cross-sectional study was conducted in the districts of Gamo Zone, Southern Ethiopia on 415 small ruminants (248 sheep and 167 goats) selected using simple random sampling technique from this region to investigate the epidemiology of gastrointestinal nematodes (GINs) and potential risk factors. Coprological examination using floatation, McMaster egg counting and larviculture were employed. The overall prevalence of gastrointestinal nematode was 68.92%. Different nematode types including strongyles (54.94%), Strongyloides papillosus. (2.65%) and Trichuris sp. (1.93%) were identified based on egg morphology with overall mean faecal egg count of 307.66 ± 38.02. Haemonchus, Trichostrongylus, Oesophagostomum, Bunostomum, Cooperia, Teladorsagia and Chabertia were identified from pooled culture. The study revealed female animals were 1.99 times (OR=1.99, 95%CI=1.16-3.41) more likely to be affected by GIN infection than males. Animals with poor (OR=2.92, 95%CI=1.32-6.45) and medium (OR=1.94, 95%CI=1.13-3.34) body condition were 2.92 and 1.94 times more likely to be affected by GINs than those with good body condition. Pregnant animals were 6.57 times (OR=6.57, 95%CI=1.49-28.89) more likely to have GINs than non-pregnant animals. Infection in young animals was 1.8 times higher (OR=1.8, 95%CI=1.05-4.2) than adults. Appropriate prevention and control methods have to be undertaken to overcome the existing problem and further study is needed to assess seasonal dynamics of the GINs.