Environmental implications of combustion of rice husk at high temperatures and for an extended period for energy generation.

The most common alternative for the management and valorization of rice processing waste is the combustion of rice husk (RH) for energy generation. The environmental risk assessment of the ash generated during the combustion of the RH to obtain energy has remained understudied. Disposal of rice husk ash (RHA) on agricultural land is the most common outcome, which could pose a risk to both natural ecosystems and human health. The objective of this study was to characterize the physicochemical composition and the phytotoxicity, cytotoxicity, and genotoxicity of RHA obtained from three distinct combustion processes. The evaluation processes were 800-900 °C in up to 5 min (I), 800-900 °C in 15-20 min (II), and 600-700 °C in 15-20 min (III). Furthermore, the content, pH, and concentrations of Al, Cd, Cu, Fe, Mg, Mn, Mo, Na, Ni, and Ti present in the ashes were determined. The germination index for two vegetable seeds was subsequently evaluated. By measuring the mitotic index and frequency of chromosomal aberrations, the cytotoxicity and genotoxicity were determined. It was observed that RHA produced by combustion of RH at higher combustion temperatures for an extended period exhibited different physicochemical properties, in addition to higher levels of phytotoxicity, cytotoxicity, and genotoxicity.

Successful lactation induction in non-pregnant gilts.

A protocol to induce lactation was applied to non-pregnant gilts. In Experiment I, five gilts with oestrus synchronized through oral supplementation of 20 mg altrenogest for 18 days received: 10 mg oestradiol cypionate (EC) on the last day of oestrous expression (D0); 10 mg EC and 300 mg long-acting progesterone (P4) on D26; and two 0.53 mg doses of a prostaglandin F2α analogue (PGF) 12 h apart on D36. Blood was collected on D12, D19, D26 and D33. Milk secretion started in all gilts 24 h after PGF administration and lasted at least 8 days. Milk samples were collected from D37 to D45. The serum P4 concentration was lower on D12 than subsequently (p < .05), but the oestradiol concentration was unaltered (p > .05). The milk produced during the induced lactation was generally richer in protein and poorer in fat compared to the milk from the lactation of a reference sow. In Experiment II, the same protocol induced lactation in two gilts, which nursed fostered piglets for 22 days. Thus, lactation was induced in all treated gilts and the milk produced was capable to nurture fostered piglets.

Paraoxonase 1 activity in the sperm-rich portion of boar ejaculates is positively associated with sperm quality.

Associations of the activity of the paraoxonase 1 (PON1) enzyme with boar sperm quality still needs to be characterized, since boar ejaculates present distinct portions with differences in sperm concentration and quality. This study evaluated PON1 activity in the serum, in the distinct portions of boar ejaculates and estimated correlations with sperm quality parameters. Ejaculates and blood samples were collected from six boars for three weeks (two per week per boar; n = 36). Serum and post-spermatic portion PON1 activities were positively correlated (P = 0.01) but were both uncorrelated with the PON1 activity in the sperm-rich portion and in the whole ejaculate (P > 0.05). Differences in PON1 activity among boars were only observed in the sperm-rich portion of the ejaculate (P < 0.05). The PON1 activity in the serum and in the post-spermatic portion was generally negatively correlated with parameters of spermatozoa kinetics (P < 0.05). In the sperm-rich portion, PON1 activity was positively correlated with sperm concentration (P < 0.0001), curvilinear distance and velocity (both P < 0.05) and DNA integrity (P < 0.05), but negatively correlated with straightness and linearity (P < 0.05). Thus, boar ejaculates with increased PON1 activity in the sperm-rich portion may present increased concentration and spermatozoa with acceptable curvilinear velocity and distance and DNA integrity, which suggests that PON1 activity may be a biomarker for potential fertility.

Removal or substitution of in feed antimicrobials in swine production.

Antimicrobial substitutes are being used in pig production systems, to maintain the health of the animals without compromising their performance. The aim of this study is to evaluate the impact of either the removal of in feed antimicrobials or their substitution for feed additives, at the nursery and growing/finishing stages. At weaning, 1091 piglets were sexed, vaccinated, homogenized by weight and allocated to six treatments during the nursery stage (26-63 d): T1- feed with no antimicrobials nor additives; T2 - feed with antimicrobials; T3 - feed with prebiotic; T4 - feed with probiotic; T5 - feed with essential oils; T6 - feed with organic acids. The same treatments were applied to 840 pigs during the growing/finishing stages (64-167 d). There was no effect of the treatments on feed conversion at the nursery (P = 0.222) and the growing/finishing (P = 0.809) stages. The average daily gain did not differ across treatments in the nursery (P = 0.342) and in growing/finishing (P = 0.050). The cost of the interventions with injectable drugs was not different between the treatments neither at the nursery (P = 0.990) nor at the growing/finishing (P = 0.310). However, the pneumonia and pleurisy index for all treatments was equal or above 1.0, which indicates a respiratory challenge. There was an increase in the cost with antimicrobials or additives per kg of feed produced, which impacts the cost per kg of pig produced. In conclusion, the removal of antimicrobials in pig diets is financially feasible and their substitution by additives did not impact growing performance.

Expression of steroidogenic enzymes and TGFß superfamily members in follicular cells of prepubertal gilts with distinct endocrine profiles.

Regulation of the transforming growth factor beta (TGFß) superfamily by gonadotrophins in swine follicular cells is not fully understood. This study evaluated the expression of steroidogenic enzymes and members of the TGFß superfamily in prepubertal gilts allocated to three treatments: 1200 IU eCG at D -3 (eCG); 1200 IU eCG at D -6 plus 500 IU hCG at D -3 (eCG + hCG); and the control, composed of untreated gilts. Blood samples and ovaries were collected at slaughter (D0) and follicular cells were recovered thereafter. Relative gene expression was determined by real-time PCR. Serum progesterone levels were greater in the eCG + hCG group compared with the other groups (P < 0.01). No differences were observed in the expression of BMP15, BMPR1A, BMPR2, FSHR, GDF9, LHCGR and TGFBR1 (P > 0.05). Gilts from the eCG group presented numerically greater mean expression of CYP11A1 mRNA than in the control group that approached statistical significance (P = 0.08) and greater expression of CYP19A1 than in both the eCG and the control groups (P < 0.05). Expression of BMPR1B was lower in the eCG + hCG treatment group compared with the control (P < 0.05). In conclusion, eCG treatment increased the relative expression of steroidogenic enzymes, whereas treatment with eCG + hCG increased serum progesterone levels. Although most of the evaluated TGFß members were not regulated after gonadotrophin treatment, the downregulation of BMPR1B observed after treatment with eCG + hCG and suggests a role in luteinization regulation.

Nanocarried antioxidants in freezing extenders for boar spermatozoa.

Post-thawing cryoinjuries in boar spermatozoa due to oxidative stress may be reduced by adding nanoencapsulated antioxidants to freezing extenders. This study evaluated post-thawing kinetics, structural and biochemical functions of boar spermatozoa frozen with extenders including resveratrol and vitamin E loaded into polymeric nanocapsules. Resveratrol was added at 0 (control), 5, 10, 20, 40 and 80 µg/ml, whereas Vitamin E was added at 0 (control), 50, 100, 200 and 400 µg/ml. Both antioxidants were tested in free and nanoencapsulated presentations. In contact with empty nanocapsules, some sperm kinetics parameters were impaired compared to the control (p < .05), whereas lipoperoxidation declined (p < .05). With inclusion of 40 µg/ml nanoencapsulated resveratrol, some sperm kinetics parameters were improved (p < .01), but sperm motility, structural and biochemical functions did not differ from the control (p > .05). No improvement in sperm quality occurred with inclusion of vitamin E, although sperm kinetics with 400 µg/ml nanoencapsulated vitamin E was reduced compared to the control (p < .01). Inclusion of 40 µg/ml nanoencapsulated resveratrol benefitted boar sperm kinetics after thawing, but no improvement resulted from inclusion of vitamin E.

Genome-wide association and genomic prediction for scrotal circumference in Hereford and Braford bulls.

Scrotal circumference (SC) is widely used as a selection criterion for bulls in breeding programs, since it is easily assessed and correlated with several desirable reproductive traits. The objectives of this study were: to perform a genome-wide association study (GWAS) to identify genomic regions associated with SC adjusted for age (SCa) and for both age and weight (SCaw); to select Tag SNPs from GWAS to construct low-density panel for genomic prediction; and to compare the prediction accuracy of the SC through different methods for Braford and Hereford bulls from the same genetic breeding program. Data of SC from 18,172 bulls (30.4 ± 3.7 cm) and of genotypes from 131 sires and 3,545 animals were used. From GWAS, the top 1% of 1-Mb windows were observed on chromosome (BTA) 2, 20, 7, 8, 15, 3, 16, 27, 6 and 8 for SCa and on BTA 8, 15, 16, 21, 19, 2, 6, 5 and 10 for SCaw, representing 17.4% and 18.8% of the additive genetic variance of SCa and SCaw, respectively. The MeSH analysis was able to translate genomic information providing biological meanings of more specific gene functions related to the SCa and SCaw. The genomic enhancement methods, especially single step GBLUP, that combined phenotype and pedigree data with direct genomic values generated gains in accuracy in relation to pedigree BLUP, suggesting that genomic predictions should be applied to improve genetic gain and to narrow the generation interval compared to traditional methods. The proposed Tag-SNP panels may be useful for lower-cost commercial genomic prediction applications in the future, when the number of bulls in the reference population increases for SC in Hereford and Braford breeds.

Prostaglandin F2α regulation and function during ovulation and luteinization in cows.

Although prostaglandins are important in the ovulation process, a precise role for prostaglandin F2α (PGF) has not been elucidated. This study aimed to evaluate the regulation of PGF receptor mRNA (PTGFR) in granulosa cells and the local effect of PGF on ovulation and luteinization. In Experiment 1, using samples collected in vivo before (Day 2), during (Day 3) and after (Day 4) follicular deviation, expression of PTGFR in bovine granulosa cells was more abundant in the dominant follicle after deviation than in subordinates (P < 0.05). However, the expression of PTGFR was not regulated (P = 0.1) in preovulatory follicles at different time-points (0, 3, 6, 12 and 24 h) after ovulation induction with GnRH. In Experiment 2, to assess the role of systemic PGF treatment on luteinization and vascularization of preovulatory follicles, flunixin meglumine (FM), a nonsteroidal anti-inflammatory drug, was used to inhibit endogenous prostaglandin synthesis. Cows with preovulatory follicles were induced to ovulate with GnRH (0 h) and allocated to three groups: Control, with no further treatment; FM, treated with 2.2 mg/kg FM im 17 h after GnRH treatment; and FM + PGF, treated with FM 17 h after GnRH, followed by 25 mg dinoprost tromethamine (PGF) 23 h after GnRH treatment. FM injection was able to reduce the concentration of PGF in the follicular fluid (FF) (P < 0.001). However, contrary to our hypothesis, color Doppler ultrasound evaluations revealed decreased vascular flow in FM + PGF group (P < 0.05), and no effect of the treatments on intrafollicular P4 and E2 concentrations 24 h after GnRH. The prostaglandin metabolite (PGFM) concentrations in the FF were greater in cows receiving systemic PGF (P < 0.001), which prompted us to further check its role on ovulation. Therefore, in Experiment 3, in a final attempt to demonstrate the local effect of PGF on ovulation, cows with preovulatory follicles received an intrafollicular injection (IFI) of PBS (Control) or 100 ng/mL purified PGF (PGF group). PGF treatment did not affect the time of ovulation after IFI (66 ± 6.4 and 63 ± 8.5 h for control and PGF, respectively; P > 0.05), further suggesting that it has no direct effect in the ovulatory process. Based on our findings, we concluded that FM decreased PGF synthesis within the follicle, whereas PGF treatment decreased follicular vascularization. In addition, the in vivo model of intrafollicular injection evidenced that PGF alone is not able to locally induce ovulation.

Phytotoxicity test in check: Proposition of methodology for comparison of different method adaptations usually used worldwide.

Seed germination tests have been widely used in recent years to indicate the toxicity levels of samples of organic compounds, biosolids, residues and effluents. Lactuca sativa L, commonly known as lettuce, has been one of the main indicative species for these tests due to its high sensitivity to low levels of toxicity, when compared to other seeds and also because it is cultivated worldwide. Although this type of analysis or essay is being widely used, it is necessary to reflect on the various adaptations of methods used by different researchers worldwide. This work presents an innovative methodology that makes it possible to compare the different phytotoxicity methods currently used in the world, through four stages that include the coefficient of variation (CV) as the main classification criterion, also counting on an eliminatory criterion. The existence of a significant difference (P value < 0.05) between the evaluated tests was proven. The phytotoxicity test that presented the lowest CV was T8 (test with lettuce seeds at 25 °C, 60 min agitation, resting overnight, 5 mL of sample on the plate, 90 mm size plate). It has concluded that not all adaptations of this type of test are reliable. It has also concluded that there is a lack of standardization for the phytotoxicity test on a global scale, which makes the various researchers in the field end up promoting variations, adaptations for the phytotoxicity test; therefore, there is an urgent need for ways to compare these variations, as the innovation proposed by this work. With a single standard methodology, we conclude that it will make it possible to compare phytotoxicity in samples directly between countries and continents, being able to generate a worldwide panorama of phytotoxicity, publicizing and comparing the standardized phytotoxicity levels in each region.

Full-Scale Composting of Different Mixtures with Meal from Dead Pigs: Process Monitoring, Compost Quality and Toxicity.

ABSTRACT: Abundant by-products of large swine industries, such as slaughterhouse sludge and carcasses, require adequate treatment to prevent negative effects of their direct disposal in the open environment. This study is aimed to evaluate the efficiency of composting using meal from dead pigs through physicochemical analyses and phytotoxic assays. Five treatments were tested, all including 50% sawdust: T1, with 50% slaughterhouse sludge (control); T2, with 20% slaughterhouse sludge and 30% meal from dead pigs; T3, with 10% slaughterhouse sludge and 40% meal from dead pigs; T4, with 20% organic stabilizing compost and 30% meal from dead pigs and T5, with 30% organic stabilizing compost and 20% meal from dead pigs. The phytotoxicity assays used lettuce, cucumber, celia, soybean, rice and wheat as bioindicators. Inclusion of meal from dead pigs was related to reduction in pH, C/N ratio, humidity and temperatures inside the pile, although thermophilic peaks lasted longer than 50 days and the final composts showed high content of nitrogen and phosphorous. The germination of bioindicators was reduced in all tested treatments, compared to the control. The composts from treatments that included meal from dead pigs presented acceptable nutrient content, which may indicate their use as organic fertilizers. However, after 4 months, all bioindicators in contact with such composts presented impaired germination. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12649-021-01422-0.

Regulation and function of leptin during ovarian follicular development in cows.

Although it is well documented that leptin signals the body nutritional status to the brain, mechanisms of leptin regulation at the ovary are not well understood. This study was conducted to determine whether there was leptin and the receptor for leptin (LEPR) in cattle ovarian follicles and to investigate potential actions of leptin on follicular growth in vivo and on regulation of granulosa cell functions in vitro. There was leptin and LEPR in granulosa and theca cells of dominant and subordinate follicles, with greater immunostaining for leptin in granulosa cells of subordinate follicles. There was a lesser relative abundance of leptin receptor gene-related protein (LEPROT) and of the adiponectin receptors 1 (ADIPOR1) and 2 (ADIPOR2) mRNA transcripts in granulosa cells of subordinate than dominant follicles (P < 0.05). Intrafollicular injection of either 100 or 1000 ng/mL leptin did not affect the diameter and the growth of dominant follicles (P> 0.05). Supplementation of in vitro culture medium with different leptin concentations did not affect (P > 0.05) the relative abundance of hydroxy-delta-5-steroid dehydrogenase, 3 beta- and steroid delta-isomerase 1 (HSD3B1), cytochrome P450 family 11 subfamily A member 1 (CYP11A1), signal transducer and activator of transcription 3 (STAT3) and X-linked inhibitor of apoptosis protein (XIAP) mRNA transcripts in granulosa cells. These findings indicate that leptin and LEPR are present in the follicular cells of cattle ovaries, but leptin apparently does not have essential functions in steroidogenesis and growth of dominant follicles.

Antibiotic resistance in microorganisms isolated in a bull semen stud.

Many microorganisms from various sources may be present in ejaculates of bulls. This study identified and isolated bacteria from bull sperm samples in a commercial stud and evaluated their resistance to antibiotics. The number of colony-forming units was determined in semen samples collected at distinct steps during freezing and thawing. The minimum inhibitory concentration and the minimum bactericidal concentration were determined for four antibiotics commonly used in commercial studs. A total of 135 microorganisms from 25 genera were isolated. After a sensitivity test, all evaluated microorganisms (n = 55) were resistant to penicillin and most of them were resistant to tylosin and lincomycin (n = 54). Resistance to all tested antibiotics was observed in 22% of all isolates, whereas only 3.9% of the isolates were inhibited by the tested antibiotics at the concentrations recommended by the international legislation. As the isolated microorganisms presented high resistance to frequently used antibiotics, sensitivity tests should be periodically conducted in commercial bull semen studs to prevent the use of contaminated semen in artificial insemination.

Safety assessment of poly(N-vinylcaprolactam) as a potential drug carrier in extenders for boar sperm cryopreservation.

Polymers may be used to deliver compounds in freezing extenders to minimize injuries in spermatozoa during cryopreservation, although their activity and toxicity for boar sperm are unknown. This study investigated the effects of the polymer (N-vinylcaprolactam) (PNVCL), when included in extenders for boar sperm cryopreservation. In Experiment 1, sperm was exposed to PNVCL at: 0 (control); 39.1; 78.1; 156.3; and 312.5 µg/mL. Spermatozoa structure, kinetics and biochemical functions were unaltered in contact with PNVCL at 38 °C (P > .05) but declined with prolonged exposure (10, 60 and 120 min) in all treatments (P > .05). In Experiment 2, after inclusion of PNVCL in the freezing extender at the same concentrations, post-thawing sperm quality did not differ compared to the control (P > .05). Lipid peroxidation and the production of reactive oxygen species were the only parameters of sperm quality that were unaffected in both experiments, even after contact with PNVCL for 120 min (P > .05). As no negative effects were observed in post-thawing boar sperm quality, PNVCL did not incur in cytotoxicity and may be a potential carrier for antioxidants in freezing extenders.

Effects of supplying omega-3 polyunsaturated fatty acids to gilts after weaning on metabolism and ovarian gene expression.

Omega-3 PUFA may benefit sow reproductive performance, but effects on weaned gilts are unknown. This study evaluated the effects of supplementing omega-3 PUFA to gilts after weaning on growth, metabolic markers, and gene expression of steroidogenic enzymes and hormone receptors. For 52 d, gilts in the control group were fed 100 g/d of regular diets, whereas gilts in the omega-3 group were fed 75 g/d of such diets plus 25 g/d of the microalgae Schizochytium sp. (3.5 g/d of omega-3 PUFA; n = 8 gilts/group). Blood samples were collected at day 0, day 21, and day 52. Total serum cholesterol levels were lower for the omega-3 group than for the control group (P < 0.05), but high-density lipoprotein-cholesterol levels were reduced at day 52 for both groups (P < 0.05). Gilts in the omega-3 group presented lower feed intake, better feed conversion, and less-intense immunolabeling for leptin and its receptor in the cytoplasm of oocytes included in primordial/primary follicles than gilts in the control group (P < 0.05). The expression of genes coding for cholesterol side-chain cleavage and aromatase enzymes and the LH receptor in follicular cells was lower for supplemented gilts (P < 0.05). Compared with controls, supplemented gilts presented decreased serum cholesterol levels and better feed conversion, but leptin presence and gene expression for steroidogenic enzymes and for the LH receptor were lower at ovarian level.

Phytotoxicity Evaluation of Wastewater from Rice Parboiling.

The aim of this study was to evaluate the phytotoxicity of raw and treated effluents from a parboiling rice industry in the south of Brazil. The evaluation included 19 physicochemical parameters and a phytotoxicity test using lettuce and cucumber seeds as bioindicators. The germination index (GI) of both seeds was decreased when in contact with the raw effluent (p < 0.05) and with the treated effluent reduced the GI of cucumber seeds (p < 0.05) and the root length (RL) of lettuce seeds (p < 0.05). For lettuce seeds exposed to the treated effluent there was a negative correlation (p < 0.05) between RL and Mn (- 0.82) and between GI and Fe (- 0.68). For cucumber seeds in the raw effluent, the Fe showed negative correlation (p < 0.05) with seed germination (- 0.75), but, in the treated effluent, their GI presented positive correlation (0.61) with the ammoniacal-N (p < 0.05). The results indicated that both effluents showed phytotoxicity for lettuce and cucumber seeds.

The implementation of a HACCP system improved the efficiency of a bull semen collection and processing center.

Bull Semen Collection and Processing Centers (SCPC) have satisfactory control of sperm quality, but commonly lack standardized quality control of hygiene procedures. This study assessed the impact of implementing a Hazard Analysis and Critical Control Points (HACCP) system in a bull SCPC, comparing microbial counts on various steps of semen processing, semen quality and costs across two periods (before and after the HACCP implementation). After surveying all routine activities of the SCPC, control points were identified, preventive measures were designed and corrective actions were employed, whenever necessary. Six months after HACCP implementation, the system was audited and production data covering two similar periods of two consecutive years were compared. Counts of colony forming units in samples collected from artificial vaginas, flexible tubes from the straw filling machine and from fresh and frozen semen after HACCP implementation were lower than during the previous period (P < 0.05). Improved post-thawing sperm motility, membrane integrity and acrosome integrity (P < 0.0001) and reduced rejection of semen batches and frozen doses were observed after HACCP implementation (P < 0.01), resulting in reduced opportunity costs. Thus, the implementation of a HACCP system in a bull SCPC allowed low-cost production of high-quality semen doses with reduced microbial contamination.

Estrus resynchronization in ewes with unknown pregnancy status.

Although fixed-time artificial insemination (FTAI) protocols are available for sheep, estrus resynchronization has not been previously reported. The objectives of this study were to evaluate the effect of estrus resynchronization with exogenous progestogen on endogenous progesterone levels and to compare pregnancy rates after two consecutive estrus synchronizations in ewes. In Experiment 1, ewes (n = 20) received an intravaginal device (IVD) containing 60 mg medroxyprogesterone acetate (MPA) for 10 days. At the IVD withdrawal (D0), ewes received 250 IU eCG and were allocated into two treatments: either no further treatment (Control; n = 10) or estrus resynchronization (Resynch; n = 10) from D12 to D19. Serum progesterone (P4) levels did not differ at D12 and D19 (P > 0.05), but were greater at D15 for the Control compared with the Resynch group (P < 0.05). In experiment 2, ewes (n = 250) were submitted to a first synchronization protocol followed by estrus detection and either artificial insemination (AI) or natural mating (NM). Subsequently, ewes were divided into two groups: Control (n = 104): which received no further treatment and were bred by NM; and Resynch (n = 146): which were submitted to a second synchronization starting on D14 (first IVD withdrawal = D0) and to NM after second IVD withdrawal (D20). Cumulative pregnancy rates did not differ between the Control (67.3%, 70/104) and Resynch (62.3%, 91/146) groups. In a third experiment, ewes (n = 83) were bred by two consecutive FTAI within a 20-day interval. Pregnancy rates after the first (30.1%, 25/83) and the second FTAI (36.2%, 21/58) did not differ (P > 0.05). In conclusion, although exogenous progestogen supplementation reduced circulating levels of P4, pregnancy maintenance was unaffected. Estrus resynchronization in ewes is feasible, resulting in similar fertility after the first and the second services. The use of resynchronization coupled with artificial insemination using semen from genetically superior rams may potentially accelerate genetic improvement in sheep herds by allowing a higher differential selection compared with natural breeding.

Phytotoxicity of effluents from swine slaughterhouses using lettuce and cucumber seeds as bioindicators.

This study evaluated the phytotoxic effects of raw and treated effluents from a swine slaughterhouse on cucumber and lettuce seeds and determined correlations among physicochemical characteristics of such effluents and the germination of seeds used as bioindicators. Physicochemical parameters were characterized for both effluents and their phytotoxicity was determined through the germination index (GI), the root length (RL) and the number of germinated seeds (SG) for both plant species. The effluents treatment system was efficient to reduce the concentration of some physicochemical parameters to levels within those recommended by the Brazilian legislation, except for P, ammoniacal N and TKN concentration. Although phytotoxicity of the treated effluent was less in comparison to the raw effluent, the GI for cucumber and lettuce seeds submitted to each of the tested effluents was lower than 80%. Thus, both effluents were phytotoxic for the tested bioindicators (p<0.05). For lettuce seeds, the GI presented negative correlations (p<0.05) with the total Kjeldahl N (-0.93) and the surfactants concentration (-0.83) in the raw effluent. The Zn concentration in the treated effluent showed a negative correlation (p<0.05) with the GI of both lettuce (-0.63) and cucumber seeds (-0.64). Therefore, effluents from swine slaughterhouses may impair the germination of the evaluated plant species if used for agricultural purposes.

Chemical castration in cattle with intratesticular injection of sodium chloride: Effects on stress and inflammatory markers.

Intratesticular injection (ITI) of sodium chloride (NaCl) is efficient for chemical castration of young calves, but its effects on calves welfare are unknown. Two experiments were conducted to evaluate the effects of ITI of 20% NaCl on stress and inflammatory markers in calves less than 20 days old and to assess the efficiency of ITI of 30% NaCl in 5 months old calves. In Experiment 1, control calves were only restrained and compared to calves submitted to castration through surgery (SC) and ITI with 20% NaCl (n = 9/group). No differences were observed for the eye corner temperature measured by thermography from 60 s before to 60 s after the procedures (P > 0.05). In the SC group, acute serum cortisol levels increased at 30 and 60 min after the procedure, but increased levels in the ITI group occurred only at 30 min (P < 0.05). Chronic discomfort markers were measured at 0, 24, 48, 72 and 96 h after the procedures (D0, D1, D2, D3 and D4, respectively). The serum levels of the paraoxonase 1 (PON1) enzyme and cortisol did not differ among groups (P > 0.05). Scrotal temperature was higher at D1 in the SC group than for the other groups, but lowest at D4 compared to the control (both P < 0.05). In Experiment 2, histological sections of testes were compared after ITI with either 30% NaCl or 30% calcium chloride (CaCl2), to intact calves (control). After 60 days, intact seminiferous tubules and mediastinum were observed after ITI with 30% NaCl, whereas coagulative necrosis, inflammatory infiltration and calcification occurred after ITI with 30% CaCl2. Efficient chemical castration through ITI of 20% NaCl in young calves was followed by slight stress and inflammatory responses compared to surgical castration. However, ITI of 30% NaCl was ineffective for chemical castration of 5 months old calves.

Selection of porcine oocytes in vitro through brilliant cresyl blue staining in distinct incubation media.

Staining with brilliant cresyl blue (BCB) may be used for oocyte selection, but BCB staining itself and the most commonly used selection medium (DMPBS) may compromise the development of porcine oocytes in vitro. This study evaluated DNA fragmentation, nuclear maturation, the area of migration of cortical granules (CG) and embryo development for stained (BCB+) and unstained (BCB-) oocytes incubated in DMPBS and in a modified medium (ReproPel) tested for the first time. Unexposed (UN), BCB+ and BCB- oocytes were incubated composing six groups: DMPBS/UN; DMPBS/BCB+; DMPBS/BCB-; ReproPel/UN; ReproPel/BCB+; and ReproPel/BCB-. There were more BCB+ oocytes in ReproPel than in DMPBS (P < 0.05). The DNA fragmentation was evaluated for oocytes in DMPBS/BCB+, DMPBS/BCB-, ReproPel/BCB+, ReproPel/BCB- and in porcine follicular fluid (control). The frequency of oocytes with no DNA fragmentation was greatest (64.6%) in DMPBS/BCB+ and lowest in ReproPel/BCB+ and ReproPel/BCB- (26.8 and 34.1%, respectively) (P < 0.05). Nuclear maturation rates were greater (P < 0.05) for DMPBS/BCB+ (63.1%), ReproPel/UN (55.1%) and ReproPel/BCB+ (50.2%) than for DMPBS/UN (40.8%) and ReproPel/BCB- (35.5%). The area of CG was greater (P < 0.05) for ReproPel/BCB- (80.7%) and DMPBS/UN (77.6%) than for ReproPel/UN (34.7%). Cleavage rates for DMPBS/BCB+ and ReproPel/BCB+ were greater than for DMPBS/UN (P < 0.05). Blastocyst development rates were greatest (P < 0.05) for ReproPel/UN and ReproPel/BCB+. In both media, BCB staining was apparently unable to select competent oocytes, which likely occurred due to toxicity. Despite the similar nuclear maturation and area of CG compared with DMPBS, oocytes selected in ReproPel presented impaired DNA integrity.