In vitro evaluation of cryopreserved bovine sperm and its relation to field fertility in fixed‐time artificial insemination

This study aimed to assess characteristics of bovine cryopreserved sperm and evaluate its relation to field fertility in fixed‐time artificial insemination (FTAI). Semen samples of 16 bulls were used to inseminate 811 Nellore cows, and four of these bulls were also used to inseminate 101 Nellore heifers. Samples of the same ejaculate used for FTAI from each bull were analysed in the laboratory after thawing. Sperm motility and vigour were subjectively assessed by light microscope, and integrity of the plasma and acrosome membranes, and H₂O₂ production were evaluated by flow cytometer. Relation among sperm characteristics and pregnancy rate of cows and heifers were evaluated by univariate and multivariate logistic regression. Subjective sperm motility and vigour did not affect the probability of pregnancy in cows or heifers. In univariate analysis for pregnancy in cows, sperm traits related to acrosome injury positively affected probability of pregnancy mainly when associated with plasma membrane integrity; H₂O₂ production seems to be less important than plasma membrane integrity in affecting probability of pregnancy. In multivariate analysis, sperm traits related to injured acrosome positively affected probability of cow and heifer pregnancies while intact acrosome was negatively related to cow pregnancy. Intact plasma membrane and high H₂O₂ production were positively related to cow pregnancy but negatively related to heifer pregnancy. Results suggest that a capacitation‐like status of the acrosome may benefit probability of pregnancy in cows.     

Effect of human tubal fluid medium and hyperactivation inducers on stallion sperm capacitation and hyperactivation

Conventional in vitro fertilization has not yet been implemented in the equine species. One of the main reasons has been the inability to develop a culture medium and incubation conditions supporting high levels of stallion sperm capacitation and hyperactivation in vitro. Although different culture media have been used for this purpose, human tubal fluid (HTF) medium, widely used in the manipulation of human and mice gametes, has not been reported so far in stallion sperm culture. The first part of this study aimed to compare HTF and Whitten's media on different stallion sperm quality and capacitation variables. Additionally, the effect of procaine, aminopyridine and caffeine in both media was evaluated on sperm motility parameters at different incubation times. Integrity and destabilization of the plasma membrane were evaluated by merocyanine 540/SYTOX Green (MC540), mitochondrial membrane potential (?Ψm) using tetramethylrhodamine methyl ester perchlorate (TMRM), acrosome membrane integrity by PNA/FITC and tyrosine phosphorylation by P‐tyrosine mouse mAb conjugated to Alexa Fluor® by flow cytometry. Motility parameters were evaluated using the integrated semen analysis system (ISAS®). We found no differences between Whitten's and HTF media and incubation time in terms of sperm viability, uninduced acrosome membrane damage or mitochondrial membrane potential at 30‐ and 120‐min incubation. Membrane fluidity (MC540) increased in both media at 30‐ and 120‐min incubation compared to noncapacitating conditions. Similarly, tyrosine phosphorylation increased in both media in capacitating conditions at 2‐ and 4‐hr incubation compared to noncapacitating conditions. Although procaine showed the best result in terms of sperm hyperactivated motility in both media, aminopyridine also showed parameters consistent with the hyperactivation including an increase in curvilinear velocity and decrease in straightness. In conclusion, HTF medium and aminopyridine equally support capacitation‐related parameters in stallion sperm.     

Haematological and biochemical analysis of healthy neonatal puppies during the immediate foetal‐to‐neonatal transition

For the neonatal patient, precocity of diagnosis is crucial for effectiveness of medical approach. However, the newborn has its own physiological peculiarities due to ongoing adaptive mechanism for extrauterine life and deserves special attention in order to underline a specific management or clinical approach. The objective of this work was to verify clinical adaptations and biochemical balance of neonates during immediate period, with special reference to haematological, renal and metabolic functions. Neonatal puppies (n = 51) were physically examined for vitality and rectal temperature at birth, 5 and 60 min post‐birth. Blood was collected at birth and 60 min post‐birth for analysis of glucose, sodium, potassium, chlorine, blood urea nitrogen (BUN), haematocrit and haemoglobin. Neonatal vitality was lower at birth compared with 5 min and 60 min post‐birth. Progressive decline in rectal temperature (36.5 ± 0.3°C, 34.2 ± 0.2°C, 32.3 ± 0.5°C) was observed at birth, 5 min and 60 min post‐birth, respectively. Puppies presented slight hyponatremia (140.7 ± 0.5 mmol/L) at birth and hypopotassemia (3.5 ± 0.1 mmol/L) and blood urea nitrogen (13.1 ± 0.7 mg/dl) during the first hour, and high haematocrit (45.1 ± 1.0%) and haemoglobin (15.3 ± 0.3 g/dl) concentration. In conclusion, puppies had rapid evolution of vitality. Marked decrease in rectal temperature occurred at 5 min post‐birth. Haematological values of neonates immediately after birth reflected mainly the dam's blood status, not being useful for a blood panel at this time point. The peculiar pattern of BUN, sodium and potassium observed during transition period, suggested that specific reference range should be considered for neonatal puppies.     

99mTc-labeled dextran for mammary lymphoscintigraphy in dogs

Lymphoscintigraphy is the technique of choice for sentinel lymph node detection in women with early breast cancer, but there is limited information evaluating the value of this technique in animals. We investigated mammary lymphatic drainage in 25 young female mongrel dogs by intramammary injection of 18.5 MBq of 99mTc-dextran (70,000 Da). Lymph node anatomical referencing was obtained using an external marker, bone scintigraphy, or scintiscanning the body contour. Cranial and caudal thoracic mammary glands drained into the cranial sternal lymph node and axillary lymph center. The cranial thoracic mammary gland also drained into the superficial cervical lymph node in two of five animals. The cranial abdominal gland was drained by the axillary lymph center. The caudal abdominal mammary gland was drained by the superficial inguinal lymph node in all animals and simultaneously by medial iliac lymph nodes in four of five animals. In one dog, this mammary gland was also drained by the mediastinal and the superficial cervical lymph nodes. The inguinal mammary gland was drained by superficial inguinal lymph nodes and simultaneously via the medial iliac lymph node in one animal. Lymphatic communications between lymph nodes were identified in 11 of 25 (44%) animals. 99mTc-dextran mammary lymphoscintigraphy was easy and rapid to perform and may provide valuable information for further studies.     

Physicochemical,Microbiological, and Sensory Properties of Pâté Elaborated from Whitemouth Croaker (Micropogonias furnieri) Muscle

The aim of this research was to formulate a unique product, croaker pâté, and to determine its physicochemical, microbiological, and sensorial characterization. Three formulations of pâté were elaborated. The pâtés fit in the stipulated regulations for the microbiological test. Regarding the texture, the pâtés presented a significant difference (p < 0.05) among each other. In the sensorial tests, the favorite pâté of the panel was formulation 3; in the acceptance test, it had higher levels of acceptance for color, flavor, and global acceptance in comparison with commercial pâté.     

Productivity of beef cattle grazing Brachiaria brizantha cv. Marandu with and without nitrogen fertilizer application or mixed pastures with the legume Desmodium ovalifolium

The use of forage legumes to contribute biologically fixed nitrogen (N) to pastures is an alternative to increase beef cattle production in tropical regions. The objective was to compare the impact of the introduction of a legume with that of N fertilizer application on forage and animal production in Brachiaria pastures. This two-year study assessed three pasture treatments: (1) mixed Marandu palisadegrass (Brachiaria brizantha [syn. Urochloa brizantha] cv. Marandu) and the legume “ovalifolium” (Desmodium ovalifolium) cv. Itabela (Mixed), (2) Marandu palisadegrass pastures with 150 kg N ha⁻¹ (Fertilized), and (3) Marandu palisadegrass without N fertilizer (Unfertilized). Rotational stocking with a variable stocking rate was used with a target herbage allowance of 1.0 kg forage kg body weight⁻¹. The pre-grazing green herbage mass was similar for Fertilized and Mixed pastures, with 54% and 63% more mass than Unfertilized pasture, respectively (p < .001). Cattle that grazed the fertilized pasture had the greatest average daily gain (ADG; p = .017). The stocking rate and liveweight gain per area were greatest for the Fertilized and Mixed pastures (p < .001 and p < .001, respectively). No differences between treatments were found for DM forage intake (p = .555). Organic matter digestibility was lowest (p < .001) for the Mixed pasture. The inclusion of the ovalifolium legume in the Marandu pasture had the same impact on beef cattle production as annual fertilization with 150 kg N ha⁻¹. The potential and environmental benefits of ovalifolium are discussed.     

For how long does the quality and quantity of residues in the soil affect the carbon compartments and CO<Subscript>2</Subscript>-C emissions?

Purpose

The mineralization/immobilization of nutrients from the crop residues is correlated with the quality of the plant material and carbon compartments in the recalcitrant and labile soil fractions. The objective of this study was to correlate the quality and quantity of crop residues incubated in the soil with carbon compartments and CO₂-C emission, using multivariate analysis.

Materials and methods

The experiment was conducted in factorial 4?+?2?+?5 with three replicates, referring to three types of residues (control, sugarcane, Brachiaria, and soybean), and two contributions of the crop residues in constant rate, CR (10 Mg ha^?1 residue), and agronomic rate, AR (20, 8, and 5 Mg ha^?1 residue, respectively, for sugarcane, soybean, and Brachiaria), evaluated five times (1, 3, 6, 12, and 48 days after incubation). At each time, we determined the CO₂-C emission, nitrogen and organic carbon in the soil, and the residues. In addition, the microbial biomass and water-soluble, labile, and humic substance carbons fractionated into fulvic acids, humic acids and humin were quantified.

Results and discussion

Higher CO₂-C emissions occurred in the soil with added residue ranging from 0.5 to 1.1 g CO₂-C m^?2 h^?1 in the first 6 days of incubation, and there was a positive correlation with the less labile organic soil fractions as well as residue type. In the final period, after 12 days of soil incubation, there was a higher relation of CO₂-C emission with carbon humin. The sugarcane and soybean residue (20 Mg ha^?1) promoted higher CO₂-C emission and the reduction of carbon residue. The addition of residue contributed to an 82.32 % increase in the emission of CO₂-C, being more significant in the residue with higher nitrogen availability.

Conclusions

This study shows that the quality and quantity of residue added to soil affects the carbon sequestration and CO₂-C emission. In the first 6 days of incubation, there was a higher CO₂-C emission ratio which correlates with the less stable soil carbon compartments as well as residue. In the final period of incubation, there is no effect of quality and quantity of residue added to soil on the CO₂-C emission.

     

Metabolic signature of spent culture media shows lipid metabolism as a determinant of pregnancy outcomes

In the present study, we investigated the spent culture media of in vitro produced (IVP) bovine embryos which did (group Pregnant) or did not (group Non-pregnant) establish pregnancy after transfer. For that purpose, IVP embryos on D5 were transferred to individual droplets for the last 48 h of culture. Embryos at the blastocyst stage were then transferred to synchronized recipients, while respective culture media drops were collected and evaluated individually. The list of metabolites present in spent culture media was obtained by electrospray ionization mass spectrometry (ESI-MS) and analysed with Metaboanalyst® to characterize the metabolic profile of each group. The spectrometric analysis showed that pathways related to lipid metabolism, particularly fatty acids degradation via beta-oxidation, were more present in the Pregnant group whereas no significant pathway was identified in the group Non-pregnant. By using this method, we were able to identify a metabolic signature in culture media that allows for a better comprehension of preferential metabolic routes taken by the most viable embryos. These findings offer great insights into the biochemistry of embryo development and reveal a potential target for the development of better-quality IVP systems, as well as tools to identify bovine embryos with greater chances to establish and maintain pregnancy.     

Importance of heat shock proteins in maize

Abiotic and biotic stress conditions cause extensive losses to maize production, mainly due to protein dysfunction in these conditions. In higher plants, the occurrence of heat-shock proteins (HSPs) in response to different environmental stresses is a universal phenomenon and has been well documented. Many studies have demonstrated that most HSPs are involved in many regulatory pathways, act as molecular chaperones for other cell proteins, and have strong cytoprotective effects. Although many functional roles for HSPs are known, the mechanisms for these multiple functions are not entirely understood. Here we reviewed the correlation among HSP genes/proteins and plant tolerance, especially maize, in different environmental stresses. Due to the low availability of information regarding the expression of HSP genes in response to different stresses in maize, we decided to mine databases in order to generate new insights related to this topic.     

Effects of aflatoxin B 1 on performance and health of tambaqui fingerlings ( Colossoma macropomum )

The detection of mycotoxins in feeds and their ingredients in aquaculture gained prominence due to losses caused in production and animal health, mainly the occurrence of aflatoxin (AFB1). The aim of this study was to evaluate the effects of AFB1 on the performance of tambaqui fingerlings (Colossoma macropomum). Four hundred tambaqui were used. Four different treatments were evaluated: treatment T1, considered as the control treatment (CT) with 3.84 μg kg−1; treatment T2, treatment T3 and treatment T4 with 500, 1000 and 2000 μg kg−1 of AFB1, respectively. The AFB1 of the samples (muscle, liver and kidney) was detected by high-pressure liquid chromatography. Four fingerlings from each treatment for histological analysis were examined. Moreover, the performance parameters (weight gain, feed conversion and feed intake) were studied. The levels of toxins used in T2, T3 and T4 represent a reduction in the growth of 14%, 35% and 45%, respectively. The T3 and T4 showed the lowest weight gain (78%) and the worst feed conversion. Aflatoxin B1 in muscle (3.28 μg kg−1) and kidneys (8.8 μg kg−1) in the T3, as well as liver (4.4 μg kg−1) and kidney (4.08 μg kg−1) in T4, was detected. Histopathological changes in liver and kidney tissues of fingerlings were more pronounced in T3 and T4. Fingerlings that consume feed contaminated with AFB1 in concentrations higher than 500 μg kg−1 present decreases in growth, reduction in weight gain and feed intake with increased feed conversion. The consumption of feed contaminated with 1000 and 2000 μg kg−1 of AFB1 caused severe deterioration in the hepatic and renal tissues.