Use of dimethylformamide to cryopreserve alpaca semen previously incubated with collagenase

The objective of this study was to evaluate the effect of collagenase and two final dimethylformamide (DMF) concentrations (4% and 7%) on alpaca frozen-thawed sperm quality. A total of 25 ejaculates from 5 alpaca were obtained using electroejaculation. Each individual ejaculate was evaluated and then diluted 4:1 in a solution of 1 mg/ml collagenase in HEPES-TALP medium and incubated for 4 min at 37°C. Subsequently, samples were diluted in TRIS-fructose-citric acid-egg yolk and cooled to 5°C. Then, each sample was divided in two aliquots and DMF at final concentration of 4% or 7% was added, equilibrated for 1 hr at 5°C and frozen over liquid nitrogen vapours. A Kruskal–Wallis test was used to evaluate the sperm morphometry, and Completely Random Block designs were used to analyse sperm motility, viability, membrane function and acrosome status. After collagenase incubation, none of the samples showed thread formation, and sperm parameters were preserved. Non-progressive motile sperm were higher (p < .05) in equilibrated samples (4% DMF: 31.8 ± 8.3% and 7% DMF: 36.3 ± 11.8%) compared to raw (10.1 ± 4.3%) and frozen-thawed semen (4% DMF: 9.7 ± 1.8% and 7% DMF: 7.5 ± 3.2%). Sperm membrane function, membrane integrity and intact acrosomes were higher (p < .05) in raw semen (40.1 ± 12.2%, 94.6 ± 3.2% and 91.3 ± 8.1%) compared to frozen-thawed samples (4% DMF: 19.8 ± 4.7%, 53.2 ± 2.7%, 65.7 ± 8.7% and 7% DMF: 20.4 ± 4.5%, 54.1 ± 1.4%, 64.6 ± 9.1%). Length of the sperm head was lower in frozen-thawed samples, being statistically different with 4% DMF compared to pre-freezing samples. The ratio between acrosome and head areas was greater (p < .05) in frozen-thawed samples. Incubation of raw alpaca semen with collagenase decreased the thread formation without affecting sperm quality. Frozen of collagenase treated alpaca semen with 4% or 7% DMF did not preserve the sperm parameters in thawed samples.     

Sperm production and quality in European eel (Anguilla anguilla) in relation to hormonal treatment

Aquaculture production relies on controlled management of gametogenesis, especially in species where assisted reproduction is needed for obtaining gametes in captivity. The present study used human chorionic gonadotropin (hCG) treatments to induce and sustain spermatogenesis in European eel (Anguilla anguilla). The aim was to evaluate effects of strip-spawning timing (12 vs. 24 hr) after weekly administration of hCG and the necessity of a primer dose (in addition to weekly hormonal treatment) prior to strip spawning (primer vs. no-primer) on sperm quality parameters. Sperm parameters included milt production (weight), density and sperm kinematics at Week 9, 11 and 13 after onset of treatment. Spermiation commenced in 11.5% of males in Week 5 and by Week 9, and all males produced milt. Male weight, milt production, sperm density and spermatocrit did not differ among hormonal treatments during the experimental period. Overall, male weight decreased from 106.3 to 93.0 g, milt weight increased from 3.5 to 5.4 g, sperm density counts decreased from 11.7 × 10⁹ to 10.5 × 10⁹ cells/ml, and spermatocrit decreased from 46.5% to 40.5%. Furthermore, spermatocrit was positively related to haemocytometer counts (R² = .86, p < .001), providing a reliable indicator of sperm density. Differences in sperm kinematics were observed depending on strip-spawning timing after hormonal injection (12 vs. 24 hr) but with no consistent pattern. These sperm quality parameters also did not consistently differ between the no-primer and primer treatments. Considering that each male may be stripped 4–5 times over the 2–3 months spawning season, omitting the primer would reduce animal handling, material costs and labour intensity, while sustaining high-quality sperm production.     

2020年底各地新玉米霉菌毒素风险分析

为了解呕吐毒素（DON）、玉米赤霉烯酮（ZEN）和黄曲霉毒素B1（AFB1）在2020年底新玉米中的污染情况,指导饲料生产企业和养殖场（户）开展霉菌毒素防控,降低霉菌毒素对饲料质量安全及养殖动物的影响,避免经济受损的风险。2020年12月在市场上采集新玉米样本225份,采用胶体金免疫层析法或上转发光免疫分析法对其霉菌毒素污染情况进行检测,依据安佑集团相关企业标准标和GB 13078-2017《饲料卫生标准》进行判定分析。结果表明：新玉米中霉菌毒素检出率为96.0%,其中DON、ZEN和AFB1的检出率分别为85.8%、87.1%和86.7%|DON、ZEN和AFB1在新玉米中的污染状况差别较大,按《饲料卫生标准》判定的超标率分别为0.0%、0.4%和11.6%|最大检测值分别为2500、534 μg/kg和＞100 μg/kg。就产地而言,山西、内蒙古、东北、安徽、山东、河南、云南、陕西等地霉菌毒素污染较重。综上,与2019年及2020年玉米霉菌毒素污染调查数据相比,2020年底新玉米污染程度较重。 [关键词] 玉米|霉菌毒素|胶体金免疫层析法|上转发光免疫分析法|污染规律     

应用液态悬浮芯片技术建立17种大肠杆菌耐药基因多重快速检测方法

为建立一种快速、高通量的多重耐药基因检测方法,利用Luminex液态芯片平台,建立了可同时检测17种耐药基因的液态芯片检测方法。该方法对大肠杆菌常见的七大类抗菌药物所对应的17种耐药基因（blaSHV、blaCMY-1、Aph3-IIa-1、aac(6)-Ib-cr、aadA-1、cmlA-1、gyrA、mcr-1、NDM-1、parC、qnrS-1、sul-1、sul-2、sul-3、tetA、tetB、tetX）进行序列分析,随后依次对其设计多重PCR特异性引物,构建特异的阳性质粒作为阳性参比品进行液态芯片检测条件优化,从而进行多重耐药基因液态芯片检测方法的研发。结果显示：成功构建了17种耐药基因的阳性质粒,并建立了两套体系用于检测17种耐药基因。在特异性试验中,两套体系中的检测信号无干扰,具有较高的特异性数值;敏感性试验中,体系一的单一质粒最低检测量为102~104 copies/μL,混合质粒为103~105 copies/μL;体系二的单一质粒最低检测量为102~105 copies/μL,混合质粒为104~106 copies/μL;有效性试验中,液态芯片法与PCR检测结果的Kappa值多在0.60以上,具有高度一致性。结果表明,本研究建立的2套液态芯片检测体系具有高通量、高灵敏和高特异性的优点,可同时对17种耐药基因进行检测,能够达到快速检测的目的。     

Free-range system and supplementation of 25-hydroxicholecalciferol increases the performance and serum vitamin levels in mixed-parity sows

Improvements in sow productivity have raised questions regarding dietary vitamin D recommendations. The present study aimed to evaluate the effects of the housing system with access to sunlight exposure and supplementation of 25-hydroxicholecalciferol on performance and serum levels of 25(OH)D₃ in sows during gestation and lactation. Sows were distributed in an experimental design with two housing systems: gestation crates or gestation free-range system with external area for sunlight exposure; and two diets: 0 or 50 μg of 25-hydroxicholecalciferol kg⁻¹. The use of 25-hydroxicholecalciferol tended (P = 0.052) to improve total born and influenced (P = 0.046) on number of born alive. Litter weight at birth was also increased (P = 0.01) by 25-hydroxicholecalciferol supplementation; 25-hydroxicholecalciferol supplementation and housing system (free-range with sunlight exposure) tended to increase weaning weight (P = 0.07) and litter daily gain (P = 0.051) during lactation. Exposure to sunlight and 25-hydroxicholecalciferol supplementation increased 25(OH)D₃ serum levels when compared with control treatment during gestation (136.95 vs. 113.92 ng mL⁻¹; P = 0.035) and lactation (120.29 vs. 88.93 ng mL⁻¹; P = 0.026). In conclusion, the association of 25-hydroxicholecalciferol supplementation with exposure to sunlight during gestation improved significantly 25(OH)D₃ serum levels and consequently performance traits in gestation and lactation.     

Ultrasound-guided modified subcostal transversus abdominis plane block and influence of recumbency position on dye spread in equine cadavers

ObjectiveTo describe and evaluate an ultrasound-guided modified subcostal approach for the transversus abdominis plane (TAP) block in horse cadavers in lateral or dorsal recumbency.Study designProspective, experimental cadaveric study.AnimalsStudy of one preserved foal and eight fresh adult horse cadavers.MethodsThe lateral and ventral abdominal wall of a preserved cadaver was dissected to identify the muscles and nerves. A unilateral standard TAP block technique was performed (60 mL of methylene blue dye–bupivacaine) on a fresh cadaver in right lateral recumbency. A modified subcostal technique was performed on the opposite side using a linear ultrasound transducer and in-plane approach. Injection points (two 30 mL dye) were at the level of the TAP (between the rectus abdominis and transversus abdominis muscles and ventral to the cutaneous trunci muscle) perpendicular to: 1) the mid-point between the xiphoid cartilage and umbilical scar; and 2) at a point between the caudal and middle thirds of the abdomen measured from the first injection point to the umbilical scar. The modified subcostal approach was performed in seven additional cadavers in both hemiabdomens, with three cadavers in lateral and four cadavers in dorsal recumbency. Ultrasound guidance was used with all injections.ResultsThe standard approach stained the sixteenth to eighteenth thoracic nerves (T16–T18). The modified subcostal approach performed in lateral recumbency provided greater spread (T9–T17) than dorsal recumbency (T12–T18) (p = 0.016).Conclusions and clinical relevanceThe modified subcostal TAP approach resulted in extensive staining exceeding the standard approach. The nerves stained are consistent with production of ventral abdominal wall anesthesia in horses. Clinical studies are needed to verify these findings.     

Effects of perioperative saphenous and sciatic nerve blocks,lumbosacral epidural or morphine–lidocaine–ketamine infusion on postoperative pain and sedation in dogs undergoing tibial plateau leveling osteotomy

ObjectiveTo compare the quality of postoperative analgesia and sedation after preoperative saphenous and sciatic nerve blockade, preoperative lumbosacral epidural injection and perioperative intravenous (IV) morphine, lidocaine and ketamine infusions in dogs undergoing stifle arthroscopy and tibial plateau leveling osteotomy (TPLO) under general anesthesia.Study designProspective, blinded, randomized, clinical comparison study.AnimalsA total of 45 dogs weighing 33.9 (15.9–56.7) kg and aged 5.2 (1.0–12.0) years, mean (range), undergoing elective unilateral TPLO for spontaneous cranial cruciate ligament rupture.MethodsClient-owned dogs were enrolled. Dogs were randomly assigned to one of three groups: group MLK, perioperative IV morphine, lidocaine and ketamine infusion; group EPID, lumbosacral epidural with ropivacaine and morphine; or group SSNB, saphenous and sciatic nerve blockade with ropivacaine. Routine stifle arthroscopy followed by TPLO surgery was performed. Sedation and pain scores were assessed at 0, 2, 4, 8 and 24 hours following extubation. Rescue analgesia was administered as prescribed by Glasgow composite pain score–short form score >5.ResultsSedation scores for MLK were higher than EPID and SSNB. Pain scores for SSNB were lower than those for EPID and MLK. No significant differences were found in anesthesia duration or surgery duration among groups. No dogs required rescue analgesia.Conclusions and clinical relevanceAlthough analgesia was adequate in all groups, the best combination of analgesia without increased sedation was recorded for SSNB.     

Questionnaire on the process of recovering horses from general anesthesia and associated personnel injury in equine practice

ObjectiveTo collect data about the current practice of recovering horses from general anesthesia and recovery personnel safety.Study designOnline survey.MethodsAn online questionnaire, including questions on general demographic data, recovery drugs, modality and characteristics of equine recovery and morbidity and mortality, was designed and distributed via e-mail to equine practitioners worldwide.ResultsPractitioners from 22 countries completed 373 questionnaires; 53% of the participants were board-certified equine surgeons, and the remainder were board-certified anesthesiologists (18%), large animal residents (8%), general practitioners (7%), large animal interns (6%), anesthesia residents (4.5%) and veterinary technicians (1.6%). Respondents were employed by academia (58%) or private practice (42%). Of the respondents employed at a university, 93% had a board-certified anesthesiologist on staff compared with 7% of respondents employed at a private practice. Most of the respondents assist horses during recovery, with 23% assisting every recovery and 44% assisting recovery in the majority of cases. Reasons for choosing to assist horses during recovery were: orthopedic procedures (57%), neurological deficits (49%), bad health (47%), history of poor recovery (44%), foals (42%), draft breeds (30%), magnetic resonance imaging (17%) and computed tomography (16%). Unacceptable recoveries were reported by 77% of participants. Commonly reported complications during recovery with any method were: orthopedic injury (66%), myopathy (54%), skin abrasion (53%) and airway obstruction (37%). The incidences of unacceptable quality of recovery (p = 0.09) or personnel injury (p = 0.56) were not different between assisted and nonassisted recoveries; however, more equine fatalities were reported for assisted recoveries (p < 0.006). Practitioners in academia reported more unacceptable recoveries (p < 0.0007) and personnel injuries (p < 0.002) compared with those in private practice.ConclusionsThe method of recovery differs among hospitals. Recovery personnel injuries associated with assisting horses during recovery are an important and previously unreported finding.     

Effect of the Nixtamalization Process on the Dietary Fiber Content,Starch Digestibility,and Antioxidant Capacity of Blue Maize Tortilla

Nixtamalization is an ancient process developed by the Mesoamerican cultures. Initially, volcanic ashes were used and then calcium hydroxide in commercial production, and more recently nixtamalization with calcium salts (NCS) has been proposed. The aim of this study was to evaluate the effect of NCS on carbohydrate digestibility and antioxidant capacity in the elaboration of blue maize tortillas. NCS in blue tortillas showed a high amount of total dietary fiber (14.27 g/100 g), the main fraction being insoluble dietary fiber. The contents of resistant starch and slowly digestible starch did not change with the nixtamalization process. The predicted glycemic index value was lower in blue tortillas with the NCS process (58) than with the traditional nixtamalization process (71). In general, NCS in blue tortillas presented a higher antioxidant capacity than traditional tortillas (ferric reducing antioxidant power method), indicating that phenolics present in blue maize maintain their activity after cooking. It can be concluded that the nutraceutical features (high dietary fiber content and antioxidant capacity) of blue maize tortillas are enhanced when they are elaborated with the NCS process.     

Effects of Annealing and Concentration of Calcium Salts on Thermal and Rheological Properties of Maize Starch During an Ecological Nixtamalization Process

The objective of the present work was to study the effect of annealing and concentration of Ca(OH)₂ (lime) and calcium salts on the thermal and rheological properties of maize starch during an ecological nixtamalization process. Thermal and rheological properties of maize starch changed during the ecological nixtamalization process because of three main causes: the annealing phenomenon, type of calcium salt, and calcium salt concentration. In all treatments thermal properties (T_o, T_p, and T_f) of nixtamal starch increased owing to the annealing process, whereas the type of salt or lime increased thermal properties and decreased pasting properties in this order: CaCl₂ > CaSO₄ > Ca(OH)₂ ≈ CaCO₃. This behavior was because of the dissociation of each salt or lime in water. Anions (OH⁻) can penetrate much more easily into the starch granule and start the gelatinization process by rupturing hydrogen bonds. Additionally, amylose‐lipid complexes were formed during the nixtamalization processes, as indicated by an increasing peak at 4.5 Å in X‐ray diffraction patterns.