Impact of Harvest Time on the Dry Matter Content, and Nutritional Parameters Related to Forage Quality of Maralfalfa (Cenchrus purpureus (Schumach.) Morrone, Poaceae) under Mediterranean Climate.

Maralfalfa (Cenchrus purpureus (Schumach.) Morrone) is a productive fodder crop in tropical regions that has been evaluated for forage nutritional value in a Mediterranean climate. To assess the nutritional value, parameters including dry matter content (DM), ash, ether extract (EE), protein (CP), fiber contents (NDF and ADF), and the amino acids profile were determined at eight harvest times (HTs) in a non-fertilized and non-irrigated crop based in Silla (Valencia, Spain). The results showed significant differences in most of the parameters studied. While CP and ash significantly decreased over the eight HTs, NDF and ADF increased. In contrast, EE and the ratio of essential amino acids/total amino acids remained constant. Values of CP remained higher than 15% during the first two HTs (16 and 28 days). According to the analyses performed, the optimum HT can be stated at 28 days as it combines high levels of CP (including an optimal combination of essential amino acids) with low levels of fibers (NDF = 57.13%; ADF = 34.76%) and a considerable amount of dry matter (15.40%). Among the essential amino acids (EA) determined, lysine and histidine showed similar values (Lys ≈ 6%, His ≈ 1.70%) when comparing the composition of these EA to other forage species and cultivars studied, whereas methionine showed lower values. This work establishes the basis for the appropriate HT of maralfalfa according to the nutritional parameters measured. Further studies could be aimed to optimize the nutritional and phytogenic properties of maralfalfa to improve its value as a fodder crop, and to finally introduce it for sustainable livestock production in Mediterranean countries.

Assisted Reproductive Technologies (ARTs) in Domestic Mammals.

As guest editors, we are pleased to present this Special Issue entitled "Assisted Reproductive Technologies (ARTs) in Domestic Mammals", comprising 10 articles of relevant interest in the field of animal reproduction [...].

Biochemical and Hematological Indexes of Liver Dysfunction in Horses.

In the present review, the authors, based on the multiple functions performed by the liver, analyze the multiple biochemical and hematological changes as an expression of altered liver function in the horse. The liver performs important metabolic functions related to the synthesis, degradation, and excretion of various substances. Modification of these functions can be evaluated and diagnosed by determining serum concentrations of several serum analytes, including enzymes and other endogenous substances. Hepatocellular enzymes, such as sorbitol dehydrogenase-SDH and glutamate dehydrogenase-GLDH, are released following hepatocellular necrosis. Hepatobiliary enzymes, such as γ-glutamyl transferase-GGT, increase in response to necrosis, cholestasis, and other alterations in bile conducts. Serum concentrations of mainly endogenous and exogenous substances that the liver should synthesize or eliminate, such as proteins (albumin and globulins), bile acids, urea, glucose, total and direct bilirubin, and coagulation factors, and fibrinogen should be included in the liver function test profile. The interpretation of laboratory tests of liver function will allow the diagnosis of functional loss of the organ. Some of the analytes considered provide information on the prognosis of liver disease. This review will provide an accurate and objective interpretation of the common biochemical and hematological tests in use in the diagnosis of equine hepatic disease patients, aiding still further the veterinary activity on the applied equine clinical cases.

Contribution of Hemogram Plan in the Horse's Clinical Evaluation.

The interpretation of the blood count is essential to help the equine clinician in the diagnosis, prognosis, patient management, and control of equine diseases. Hematologic alterations often reflect the condition of the individual or an overall response to a pathological situation. A thorough clinical examination of the patient is essential to correctly interpret the hematological results. The most common abnormalities in the erythrogram are mainly anemia and polycythemia. The frequent causes of anemia in horses are acute and chronic blood loss, hemolytic anemia, and anemia caused by chronic disease. Evaluation of leukogram, including a total white cell count, a differential cell count, absolute numbers of specific leukocytes can help identify abnormalities that may suggest specific diseases such as a viral or bacterial infection, inflammatory disorders or even a neoplastic process. The platelet count is most often used to monitor or diagnose conditions that cause too much bleeding related with thrombocytopenia; it can be due to multiple mechanisms such as reduction of thrombopoiesis (myeloptisis, myelofibrosis, myeloproliferative disease, and idiopathic medullary aplasias or due to the effect of mielosuppresive drugs), increased peripheral destruction of platelets (immune mediated thrombocytopenia), consumption (dissemined intravascular coagulation) sequestration of the spleen and loss of platelets by idiopathic origin.

Analysis of Uterine Blood Flow in Breeding Sows through the Estrus and Early Diestrus, and after Artificial Insemination.

This study aimed to determine uterine blood flow indices by transabdominal Doppler ultrasound in sows (n = 18) under different conditions: (i) sows after estrus detection (day 0, D0); (ii) sows 2 h after artificial insemination (AI), performed 24 h after detection of estrus (day 1, D1); (iii) sows in early diestrus (day 5, D5). Moreover, three different types of seminal doses were used for AI depending on the ejaculate fraction included (F1: doses containing only the rich fraction of the ejaculate; F2: F1 + the transition fraction between rich and poor fractions; F3: F2 and poor fraction). The statistical analysis revealed significant differences in some indices regarding the period of analysis (D0, D1, and D5). Diastolic velocity and mean velocity showed lower values at D5 in comparison with D0 and D1 (p < 0.01). On the other hand, the pulsatility index and the relationship systolic velocity/diastolic velocity indicated higher values at D5 in comparison with D0 and D1 (p < 0.01). No differences were observed regarding the type of seminal dose used in any of the time points analyzed (p > 0.05). Neither insemination per se nor the type of ejaculate fraction used immediately modified the uterine vascularity, but some indices are affected by the stage of the estrus cycle (estrus vs. early diestrus).

Hepatic Enzyme Profile in Horses.

For diagnostic purposes, liver enzymes are usually classified into hepatocellular and cholestatic. These two groups of equine liver-specific enzymes include sorbitol dehydrogenase (SDH), glutamate dehydrogenase (GLDH), γ-glutamyl transferase (GGT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), and alkaline phosphatase (ALP). SDH and GLDH mostly reflect hepatocellular injury and cholestasis, while GGT expresses high values in biliary necrosis or hyperplasia. Likewise, AST, LDH, and ALP also reflect hepatocellular and biliary disease, but these enzymes are not liver specific. From the clinical point of view of the course of liver or biliary disease, AST and ALP are indicative of chronic disease, whereas SDH, GGT, and GLDH indicate an acute course. The patterns of enzymatic changes at the blood level are associated with different types of liver pathologies (infectious, inflammatory, metabolic, toxic, etc.). Increases in hepatocellular versus biliary enzyme activities are indicative of a particular process. There are different ways to diagnose alterations at the hepatic level. These include the evaluation of abnormalities in the predominant pattern of hepatocellular versus cholestatic enzyme abnormalities, the mild, moderate, or marked (5−10-fold or >10-fold) increase in enzyme abnormality concerning the upper limit of the reference range, the evolution over time (increase or decrease) and the course of the abnormality (acute or chronic).

MOET Efficiency in a Spanish Herd of Japanese Black Heifers and Analysis of Environmental and Metabolic Determinants.

Multiple ovulation and embryo transfer (MOET) systems have been intensively implemented in Japanese Black cattle in Japan and to create Japanese Black herds out of these areas. Environmental conditions influence MOET efficiency. Thus, we describe results of 137 in vivo, non-surgical embryo flushings performed between 2016-2020, in a full-blood Japanese Black herd kept in Spain and the possible effects of heat, year, bull, donor genetic value, and metabolic condition. Additionally, 687 embryo transfers were studied for conception rate (CR) and recipient related factors. A total of 71.3% of viable embryos (724/1015) were obtained (5.3 ± 4.34/flushing). Donor metabolites did not affect embryo production (p > 0.1), although metabolite differences were observed over the years, and by flushing order, probably related to the donor age. CR was not affected by embryo type (fresh vs. frozen), recipient breed, and whether suckling or not suckling (p > 0.1). CR decreased significantly with heat (44.3 vs. 49.2%; (p = 0.042)) and numerically increased with recipient parity and ET-number. Pregnant recipients showed significantly higher levels of cholesterol-related metabolites, glucose, and urea (p < 0.05). Therefore, adequate MOET efficiency can be achieved under these conditions, and heat stress should be strongly avoided during Japanese Black embryo transfers. Moreover, recipients' metabolites are important to achieve pregnancy, being probably related to better nutrient availability during pregnancy.

A review of current knowledge of myeloproliferative disorders in the horse.

Myeloid disorders are conditions being characterized by abnormal proliferation and development of myeloid lineage including granulocytes (neutrophils, eosinophils and basophils), monocytes, erythroids, and megakaryocytes precursor cells. Myeloid leukemia, based on clinical presentation and proliferative rate of neoplastic cells, is divided into acute (AML) and myeloproliferative neoplasms (MPN). The most commonly myeloid leukemia reported in horses are AML-M4 (myelomonocytic) and AML-M5 (monocytic). Isolated cases of AML-M6B (acute erythroid leukemia), and chronic granulocytic leukemia have also been reported. Additionally, bone marrow disorders with dysplastic alterations and ineffective hematopoiesis affecting single or multiple cell lineages or myelodysplastic diseases (MDS), have also been reported in horses. MDSs have increased myeloblasts numbers in blood or bone marrow, although less than 20%, which is the minimum level required for diagnosis of AML. This review performed a detailed description of the current state of knowlegde of the myeloproliferative disorders in horses following the criteria established by the World Health Organization.

Supplementation of the thawing medium with reduced glutathione improves function of frozen-thawed goat spermatozoa.

Sperm cryopreservation represents a useful tool in the management of reproduction in goat production. However, freezing and thawing produce physical and chemical stress on the sperm membrane that reduces their viability and fertilizing ability. In this study, firstly we evaluated the effects of reduced glutathione (GSH, 1 and 5mM) supplementation of the thawing extender on parameters of frozen-thawed goat spermatozoa. We used a set of functional sperm tests that included sperm motility assayed by computer-assisted semen analysis (CASA), membrane lipid packing disorder, spontaneous acrosome reaction, free radical production (ROS generation) and sperm chromatin condensation. The main findings from this study were that addition of GSH to the thawing medium resulted in: (1) a higher motility and progressive motility; (2) a higher number of non-capacitated viable spermatozoa; (3) higher number of viable spermatozoa with intact acrosome; (4) a reduction in ROS generation and (5) lower chromatin condensation. In a second study, the additions of reduced (GSH, 5mM) or oxidized glutathione (GSSG, 2.5mM) to the thawing media were evaluated. We confirmed the protective effect of GSH on the sperm functionality. The addition of GSSG to the thawing media was less protective to sperm functions compared to GSH. Addition of GSH to the thawing extender could be of significant benefit in improving the function and fertilizing capacity of frozen goat spermatozoa. The information derived from this study suggests the importance of oxidative stress as responsible for cryo-injury to spermatozoa and opens new windows to explore the practical application of antioxidants to improve the quality of post-thaw goat semen.

Supplementation of the dilution medium after thawing with reduced glutathione improves function and the in vitro fertilizing ability of frozen-thawed bull spermatozoa.

In this study, we evaluated the effects of glutathione (l-gamma-glutamyl-l-cysteinylglycine; GSH) supplementation of the thawing extender on bull semen parameters to compensate for the decrease in GSH content observed during sperm freezing. To address these questions fully, we used a set of functional sperm tests. These included tests of sperm motility assayed by computer-assisted semen analysis, membrane lipid packing disorder, spontaneous acrosome reaction, free radical production [reactive oxygen species (ROS) generation], sperm chromatin condensation, DNA fragmentation by terminal deoxynucleotidyltransferase-mediated dUTP nick-end labelling and acridine orange staining measured by flow cytometry. Finally, the in vitro penetrability of in vitro matured oocytes and the in vitro production of embryos were evaluated. The main findings emerging from this study were that addition of GSH to the thawing medium resulted in: (i) a higher number of non-capacitated viable spermatozoa; (ii) a reduction in ROS generation; (iii) lower chromatin condensation; (iv) lower DNA fragmentation; (v) higher oocyte penetration rate in vitro and (vi) higher in vitro embryo production compared with control group. Nevertheless, GSH had no significant effect on motion parameters or the occurrence of the spontaneous acrosome reaction. Addition of GSH to the thawing extender could be of significant benefit in improving the function and fertilizing capacity of frozen bull spermatozoa.

Effect of estrus synchronization and artificial insemination on reproductive performance of Merino sheep

The experiment was conducted to determine the reproductive performance (lambing rate and litter size) of sheep artificially inseminated at a synchronized estrus in relation to rams, dose of PMSG and number of artificial inseminations performed. During spring cyclic Merino adult ewes were treated with intravaginal sponges impregnated with 60 mg medroxyprogesterone acetate (MAP). After 14 days sponges were removed and the females received an intramuscular injection of either 375 or 400 IU pregnant mare serum gonadotropin (PMSG). Estrus detection was performed with vasectomized rams. Females in estrus were cervically inseminated with fresh diluted semen from different rams either once or twice. Data on lambing rate and litter size were recorded. Lambing rate was not affected by rams or number of inseminations. A higher proportion of ewes that received 400 IU PMSG lambed compared with those receiving 375 IU PMSG (76.47 percent v 54.32 percent; p<0.01). Litter size was not affected by any of the variables under study

Estrus presentation and distribution in ewes treated with intravaginal sponges impregnated with medroxyprogesterone acetate (MAP) in combination with pregnant mare serum gonadotropin (PMSG)

The objectives of this study were: 1) to determine estrus presentation and distribution following a conventional method of estrus synchronization (progestagen-PMSG treatment) in an ewe herd and 2) to analyze estrus presentation and distribution in adult ewes and ewe lambs. During spring a total of 300 cyclic Merino ewes, including 231 adult ewes and 69 ewe lambs were treated with intravaginal sponges impregnated with 60 mg medroxyprogesterone acetate (MAP). After 14 days sponges were removed and 375 IU pregnant mare serum gonadotropin (PMSG) were administered i.m. Estrus detection was performed with vasectomized rams. Ewes were inspected for the presence of marks at 4-hours intervals. Sponge losses, estrus synchronization and distribution were analyzed for adult ewes and ewe lambs. It was detected 1 per cent (3/300) of sponge losses. Estrus synchronization rate was 92.93 per cent (276/297) for the ewe herd, being 93.48 per cent (215/230) for adults and 91.04 per cent (61/67) for lambs (p>0.10). Estrus onset was detected from 28 to 68 hours following treatment in both classes of females. The interval between sponge removal and estrus onset was 46.88 ñ 11.78 hours for the ewe herd, being 46.99 ñ 12.22 hours for adult ewes and 47.31 ñ 10.94 hours for ewe lambs (p>0.10). Statistical differences were found only for the intervals 34-38 (p<0.10) and 50-54 hours (p<0.05) between adult ewes and ewe lambs. It was concluded that the treatment used was effective for estrus synchronization in ewes

Residual levels on medroxyprogesterone acetate (MAP) impregnated sponges after estrus synchronization treatment in cyclic ewes

A pool of polyurethane sponges impregnated with medroxyprogesterone acetate (MAP) was prepared. Real level of progestagen on sponges was checked prior to sponge treatment insertion. During spring a total of 126 cyclic Merino ewes were intravaginally treated with MAP-impregnated pessaries for the synchronization of estrus. After 14 days treatment, sponges were removed. Estrus detection was performed with vasectomized rams. Ewes were inspected for the presence of marks twice a day. Ewes in estrus were artificially inseminated with fresh diluted semen containing a dose of 300x106 total sperm. Artificial insemination was performed 12 hours after estrus presentation. Residual levels of MAP (RMAP) on removed sponges were measured by spectrophotometry at 241 nM and examined in relation to their estrus response and fertility. Real dose of MAP was in average 54 mg. RMAP found on sponges following treatment were in average 25.00 ñ 0.84 mg. Percentage of estrus synchronization was 92.86 per cent and pregnancy rate was 50.43 per cent. There were no significant differences for RMAP between ewes with (24.70 ñ 0.86 mg) and without (28.89 ñ 3.65 mg) estrus response (p>0.10). There were also no significant differences for RMAP between pregnant (25.56 ñ 1.25 mg) and non-pregnant (23.83 ñ 1.18 mg) ewes (p>0.10). It was concluded that the 60 mg MAP dose conventionally used for estrus synchronization in ewes is higher than the amount uptaken by the females