Gene expression and immunolocalization of fibroblast growth factor 2 in the ovary and its effect on the in vitro culture of caprine preantral ovarian follicles

This study quantified Fibroblast growth factor 2 (FGF-2) mRNA and localized FGF-2 protein in different categories of follicles isolated from goat ovaries. In addition, we verified the effects of this factor on the in vitro culture of preantral follicles isolated from goats. For mRNA quantification, we performed real-time PCR using primordial, primary and secondary follicles, as well as cumulus-oocyte complexes (COCs) and mural granulosa and theca cells of small and large antral follicles. For FGF-2 protein localization, the ovaries were subjected to conventional immunohistochemical procedures. Preantral follicles were isolated and cultured in vitro for 12 days in either control (basic) or supplemented with FGF-2 medium. The expression of FGF-2 mRNA was detected in all categories of follicles and there was no difference in preantral follicles and COCs or granulosa/theca cells from small and large antral follicles. However, in large antral follicles, COCs showed expression levels significantly lower than in granulosa/theca cells (p < 0.05). We observed moderate expression of FGF-2 protein in preantral follicles but not in granulosa cells of primordial follicles and theca cells of secondary follicles. In both small and large antral follicles, strong, moderate and weak staining was observed in oocytes, granulosa and theca cells, respectively. The addition of FGF-2 caused a significant increase in the daily follicular growth rate compared to the control group. We conclude that FGF-2 mRNA is expressed throughout follicular development and that its protein can be found in different patterns in preantral and antral follicles. Furthermore, FGF-2 increases the follicular growth rate in vitro.     

The effect of LIF in the absence or presence of FSH on the in vitro development of isolated caprine preantral follicles

We investigated the effect of the leukaemia inhibitory factor (LIF) alone or in association with FSH on the in vitro culture (IVC) of caprine preantral follicles. Preantral follicles >200 μm in size were isolated and cultured for 18 days in basic medium either alone (control) or supplemented with LIF (10 or 50 ng/ml) in the absence or presence of FSH. Every 6 days, follicular survival, growth and antrum formation were evaluated. At the end of the culture period, the oocytes underwent in vitro maturation (IVM), and their viability and chromatin configuration were assessed. Follicles of the control group and those cultured in 10 ng/ml LIF maintained the structural integrity (particularly the preservation of the basement membrane) when compared to the oocytes cultured in 50 ng/ml LIF, regardless the presence of FSH. In the absence of FSH, the percentage of antrum formation after 18 days of culture in the 50 ng/ml LIF group was significantly lower than in either the control group or the 10 ng/ml LIF group. However, this effect was not observed in the presence of FSH. The rate of resumption of meiosis was significantly higher in the 50 ng/ml LIF group in the absence of FSH in comparison with the control and 10 ng/ml LIF groups. Metaphase II was observed only when follicles were cultured in a combination of FSH and 50 ng/ml LIF. In conclusion, LIF alone does not interfere with antral formation and oocyte growth, but at concentration of 50 ng/ml and combined with FSH, it promotes oocyte maturation.     

Interaction between melatonin and follicle-stimulating hormone promotes in vitro development of caprine preantral follicles

The aim of this study was to investigate the effects of melatonin and follicle-stimulating hormone (FSH) on the in vitro culture of goat preantral follicles. Ovarian fragments were cultured for 7 d in α-minimum essential medium (α-MEM⁺) containing melatonin (100, 250, 500, or 1,000 pM), FSH (50 ng/mL), or a combination of the 2 hormones and further analyzed by histology and transmission electron and fluorescent microscopy. The results showed that after 7 d of culture, tissues cultured in α-MEM⁺ alone or supplemented with FSH alone, melatonin (500 and 1,000 pM), or the combination of FSH and melatonin (1,000 pM) maintained percentages of normal preantral follicles similar to the fresh control. In contrast to the noncultured tissues, the percentage of developing follicles was increased under all culture conditions after 7 d (P < 0.05). The addition of 1,000 pM melatonin associated with FSH to the culture medium increased follicular and oocyte diameters compared with α-MEM⁺ alone after 7 d of culture (P < 0.05). Ultrastructural and fluorescent analyses confirmed the integrity of follicles cultured with 1,000 pM of melatonin plus FSH for 7 d. In conclusion, this study demonstrated that the interaction between melatonin and FSH maintains ultrastructural integrity and stimulates further growth of cultured caprine preantral follicles.     

Diversity of non‐structural carbohydrates in grasses (Poaceae) from Brazil

Non‐structural carbohydrates (NSC) are the primary energy available for growth and dry‐mass production in forage grasses. They are also associated with tolerance to environmental stresses, including drought. Soluble carbohydrates, especially fructans of temperate forage grasses, have been extensively studied; however, little is known about NSC of tropical grasses. These plants are abundant in the Cerrado, a savanna‐like vegetation with a definite seasonality in rainfall distribution. Such an environment presents an ideal condition for the occurrence of fructans. However, the present analysis of carbohydrates in aerial parts of twenty‐four tropical grass species, mainly from the Panicoideae subfamily in the Cerrado, shows a distinctively different NSC profile when compared with temperate grasses. Free glucose and starch are common NSC found in most plant organs. However, free fructose and sucrose‐based oligosaccharides were only found in very low amounts. Sucrose was detected in higher amounts only in stems. A linear series of malto‐oligosaccharides was found in leaf blades, sheaths and stems of most of the Cerrado grasses, except for those from the Andropogoneae tribe. These results indicate a diversity of regulatory pathways of NSC metabolism in tropical forage grasses from the Cerrado.     

Mixed-effect non-linear modelling for diameter estimation along the stem of Tectona grandis in mid-western Brazil

This study evaluated the efficiency of taper functions and the application of mixed-effect modelling for diameter estimation along the stems of Tectona grandis. We sampled 266 trees of Tectona grandis, measuring the diameter at relative heights for volume determination, grouping the data according to three form-factor classes. Six taper functions were fitted, selecting the function with better fit performance. Six taper functions were fitted, selecting the function with better fit performance. The selected function was fitted in its basic formulation, and with the mixed non-linear modelling technique in different scenarios, and for the stem stratified in three portions of the total height. The precision and selection of the adjusted models were evaluated regarding the coefficient of determination, standard error of estimate, the Akaike information criterion, bias, quadratic error and absolute bias. According to the statistical criteria used, the model of Kozak was selected for the adjustments. For diameter estimation, the scenario with two coefficients as random effects provided an accuracy increase of 11.91%, and the mixed non-linear modelling better estimated the stem diameter for the stratified stems. In conclusion, the model of Kozak can be used to describe the stem shape of Tectona grandis, and the mixed-effect non-linear model approach was the best technique to estimate diameter along the stem of Tectona grandis.     

Are Pseudokirchneriella subcapitata and Chlorella vulgaris Affected by Environmental Samples from a Rice Field?

Rice fields represent important production rates in Portugal. However, the intensive soil management and the exploitation of agrochemicals may pose a threat to nontarget organisms. Hence, the present work regards the toxicity screening of surface waters and sediment elutriates collected during the drainage of fields in the vicinity of a rice paddy (Quinta do Seminário, Soure, Portugal): 1. in River Pranto (RP), the river from which the field irrigation water is canalized; 2. inside the rice paddy, from the main drainage channel – Vala de Enxugo (VE). For that purpose, it was used a combination of physico-chemical analyses and bioassays with two green algae species – Pseudokirchneriella subcapitata and Chlorella vulgaris. The chemical screening showed an apparent absence of xenobiotics in sediment samples, while no pesticides were found within the chemical contaminants detected in water samples. The nutrient load reflected low levels of inorganic contamination. Bioassays revealed that P. subcapitata was more sensitive to the overall physico-chemical conditions in natural samples than C. vulgaris, being its growth inhibited under water samples from both sites. On a whole, water samples, mainly those from the main irrigation/drainage channel of the rice fields (VE), were more deleterious to microalgae than those from RP or any of the elutriates.     

Assessment of brachial plexus blockade in chickens by an axillary approach

ObjectiveTo assess the brachial plexus block in chickens by an axillary approach and using a peripheral nerve stimulator.Study designProspective, randomized, double-blinded study.AnimalsSix, 84-week old, female chickens.MethodsMidazolam (1 mg kg⁻¹) and butorphanol (1 mg kg⁻¹) were administered into the pectoralis muscle. Fifteen minutes later, the birds were positioned in lateral recumbency and following palpation of the anatomic landmarks, a catheter was inserted using an axillary approach to the brachial plexus. Lidocaine or bupivacaine (1 mL kg⁻¹) was injected after plexus localization by the nerve stimulator. Sensory function was tested before and after blockade (carpus, radius/ulna, humerus and pectoralis muscle) in the blocked and unblocked wings. The latency to onset of motor and sensory block and the duration of sensory block were recorded. A Friedman nonparametric one-way repeated-measures anova was used to compare scores from baseline values over time and to compare the differences between wings at each time point.ResultsA total of 18 blocks were performed with a success rate of 66.6% (12/18). The latency for motor block was 2.8 ± 1.1 and 3.2 ± 0.4 minutes for lidocaine and bupivacaine, respectively. The latencies for and durations of the sensory block were 6.0 ± 2.5 and 64.0 ± 18.0 and 7.8 ± 5.8 and 91.6 ± 61.7 minutes for lidocaine and bupivacaine, respectively. There was no statistical difference between these times for lidocaine or bupivacaine. Sensory function was not abolished in nonblocked wings.Conclusions and clinical relevanceThe brachial plexus block was an easy technique to perform but had a high failure rate. It might be useful for providing anesthesia or postoperative analgesia of the wing in chickens and exotic avian species that have similar wing anatomy.     

Strategies of the control of an outbreak of leptospiral infection in dairy cattle in Northeastern Brazil

The aim of the present study was to describe the strategies of the control of an outbreak of leptospiral infection in dairy cattle in Maranhão State, Northeastern Brazil. In the period from January to July 2015, 18 (17%) out of 106 cows presented abortion, six (5.7%) stillbirth, and 12 (11.3%) repeated estrus, totaling 24 animals with reproductive problems. The diagnosis of leptospirosis was based on serology (microscopic agglutination test—MAT), bacteriological culture, and polymerase chain reaction (PCR). Antibiotic therapy, vaccination protocols, and changes in management practices were suggested as control measures. Of all animals on the farm (n?=?280), 136 (48.6%) were seropositive for at least one serovar of Leptospira sp. No pure leptospiral culture was obtained. Eight of the animals with reproductive problems yielded positive PCR results (vaginal fluid of seven animals and urine and vaginal fluid of one animal). Genetic sequencing of a vaginal fluid/urine PCR-positive sample revealed Leptospira borgpetersenii. One year after the adoption of control measures, no reproductive problems were observed. Thus, leptospirosis probably caused the reproductive failures in the herd, and the control and prevention measures implemented were efficient in controlling the disease.

     

The phenotypic relationship between residual intake and gain and other feed efficiency traits in Nellore cattle

This study aimed to compare feed efficiency measures of Nellore beef cattle on different residual intake and gain (RIG) classes. We used data from 610 animals weighing on average 236.33 kg and average of 283 days of age from feedlot performance tests carried out between 2005 and 2012. Animals were grouped based on RIG into three different classes: high RIG (>?mean?+?0.5 standard deviation (SD), most efficient; n?=?193), medium RIG (mean?±?0.5 SD; n?=?235), and low RIG (<?mean – 0.5 SD, least efficient; n?=?182). Residual feed intake (RFI), residual gain (RG), feed conversion ratio (FCR), feed efficiency (FE), relative growth rate (RGR), and Kleiber ratio (KR) of animals in each RIG class were compared by Tukey test at 1% of probability. Phenotypic correlations between variables were evaluated as well. Animals on high RIG class showed lower dry matter intake (P?<?0.01) and higher average daily gain (P?<?0.01) than low RIG animals. Consequently, high RIG animals had lower FCR (P?<?0.01) and higher FE (P?<?0.01) than those animals in low RIG class. The most efficient animals based on RIG were also the most efficient animals based on RG and RFI. RIG was negatively correlated to dry matter intake (P?<?0.01) and FCR (P?<?0.01), and a positive correlation was found between RIG and FE (P?<?0.01). Therefore, RIG appears to be a good parameter to select animals with reduced dry matter intake and high productive performance.