Enzyme Bioprospection of Marine-Derived Actinobacteria from the Chilean Coast and New Insight in the Mechanism of Keratin Degradation in Streptomyces sp. G11C

Marine actinobacteria are viewed as a promising source of enzymes with potential technological applications. They contribute to the turnover of complex biopolymers, such as pectin, lignocellulose, chitin, and keratin, being able to secrete a wide variety of extracellular enzymes. Among these, keratinases are a valuable alternative for recycling keratin-rich waste, which is generated in large quantities by the poultry industry. In this work, we explored the biocatalytic potential of 75 marine-derived actinobacterial strains, focusing mainly on the search for keratinases. A major part of the strains secreted industrially important enzymes, such as proteases, lipases, cellulases, amylases, and keratinases. Among these, we identified two streptomycete strains that presented great potential for recycling keratin wastes—Streptomyces sp. CHA1 and Streptomyces sp. G11C. Substrate concentration, incubation temperature, and, to a lesser extent, inoculum size were found to be important parameters that influenced the production of keratinolytic enzymes in both strains. In addition, proteomic analysis of culture broths from Streptomyces sp. G11C on turkey feathers showed a high abundance and diversity of peptidases, belonging mainly to the serine and metallo-superfamilies. Two proteases from families S08 and M06 were highly expressed. These results contributed to elucidate the mechanism of keratin degradation mediated by streptomycetes.     

First results of embryonic development,spawning and larval rearing of the Mediterranean spider crab Maja squinado (Herbst ) under laboratory conditions,a candidate species for a restocking program

The Mediterranean spider crab, Maja squinado, is depleted due to overfishing. The crab has virtually disappeared from areas where it was abundant, such as the Balearic Islands and the Catalan coast. Maja squinado, is economically and ecologically very valuable, and it is essential to obtain information on its biology and rearing conditions to attempt to repopulate the damaged stocks of the species in the Mediterranean basin. Herein, we describe the first successful rearing of M. squinado under laboratory conditions. Our results show that M. squinado is an excellent candidate for restocking using cultured juveniles. Two consecutive broods with a 1–4 day interbrood period were observed in the laboratory in wild‐caught females, the maximum observed duration of embryonic development of the egg mass being 32 days at 18.4 ± 0.9°C, and went through four different stages. The complete larval and first juvenile development was studied in laboratory cultures fed enriched Artemia metanauplius. At 19.6 ± 0.6°C, development from hatching to first crab moult took 17 days, and it comprised two zoeae stages and one megalopa stage. The survival rate at the different stages was monitored, and 7.13 ± 2.3% was achieved at the first crab instar.     

Longitudinal growth strain as a log and wood quality predictor for plantation-grown <Emphasis Type="Italic">Eucalyptus nitens</Emphasis> sawlogs

Peripheral longitudinal growth strain (LGS) was measured on a total of 81, 22-year-old pruned Eucalyptus nitens trees from five thinning treatments in a plantation thinning trial in Tasmania. Growth strain data were gathered at breast height on each tree using the CIRAD-Forêt method. The effects of thinning treatment and tree diameter on LGS and its relationships to solid-wood traits determined in a processing trial were examined. LGS was significantly higher in the direction of the prevailing wind and was not significantly affected by either thinning treatment or tree diameter. An index of log end splitting was positively related to tree diameter, sawlog position (upper/lower), and LGS, which explained up to an additional 20% of the variance after tree diameter and log position were accounted for in a linear regression model. High LGS and log end splitting were significant indicators of increased board end splitting.     

Influence of components of infant formulas on in vitro iron, zinc, and calcium availability

The effect of casein content and Ca concentration on Fe, Zn, and Ca dialyzability was assessed using a response surface design. Tested casein levels were 5.31-13.75 g/L (34.8-90.2% of total protein). Whey protein was added to complete 15.25 g/L total protein. Calcium levels were adjusted with calcium citrate within a range between 417.4 and 804.9 mg/L. Through the experimental design utilized, we found that of both assessed factors, only the casein content significantly influenced Fe and Zn dialyzability. Protein composition did not influence calcium dialyzability, and calcium concentration did not affect either Fe or Zn dialyzability. No effect of casein-Ca on iron, zinc, and calcium dialyzability was found. According to these results, whey-dominant formulas are less prone to hamper mineral availability, and are therefore suitable in order to improve iron and zinc availability.     

Validation of the FAMACHA eye color chart for detecting clinical anemia in sheep and goats on farms in the southern United States

Recent studies on sheep and goat farms in the southern United States indicate that multiple-anthelmintic resistance in Haemonchus contortus is becoming a severe problem. Though many factors are involved in the evolution of resistance, the proportion of the parasite population under drug selection is believed to be the single most important factor influencing how rapidly resistance develops. Therefore, where prevention of resistance is an important parallel goal of worm control, it is recommended to leave a portion of the animals untreated. Recently, a novel system called FAMACHA was developed in South Africa, which enables clinical identification of anemic sheep and goats. When H. contortus is the primary parasitic pathogen, this system can be applied on the farm level to reduce the number of treatments administered, thereby increasing the proportion of the worm population in refugia. Since most studies validating the FAMACHA method have been performed in South Africa, it is important that the method be tested in other regions before its use is broadly recommended. We performed a validation study of FAMACHA by testing the system in sheep (n = 847) and goats (n = 537) of various breeds and ages from 39 farms located in Arkansas, Georgia, Louisiana, Florida, and the US Virgin Islands. The color of the ocular conjunctiva of all animals were scored on a 1-5 scale using the FAMACHA card, and blood samples were collected from each animal for determination of packed cell volume (PCV). Fecal samples were also collected from a majority of the animals tested for performance of fecal egg counts (FEC). Correlations between PCV and eye scores, PCV and FEC, and FEC and eye scores were all highly significant for both sheep and goats (P < 0.001). Data for both FAMACHA scores and PCV were evaluated using two separate criteria for anemia: eye score values of 3, 4 and 5 or 4 and 5, and PCV values of < or =19 or < or =15 were considered anemic. Specificity was maximized when eye score values of 4 and 5 were considered anemic and PCV cut off for anemia was < or =19, but sensitivity was low. In contrast, sensitivity was 100% for both sheep and goats when eye score values of 3, 4 and 5 were considered anemic and PCV cut off was < or =15, but specificity was low. In both sheep and goats, predictive value of a negative was greater than 92% for all anemia and eye score categories, and was greater than 99% for both eye score categories when an anemia cutoff of < or =15 was used. Predictive value of a positive test was low under all criteria indicating that many non-anemic animals would be treated using this system. However, compared to conventional dosing practices where all animals are treated, a large proportion of animals would still be left untreated. These data indicate that the FAMACHA method is an extremely useful tool for identifying anemic sheep and goats in the southern US and US Virgin Islands. However, further studies are required to determine optimal strategies for incorporating FAMACHA-based selective treatment protocols into integrated nematode control programs.     

Effect of oxytocin infusion on luteal blood flow and progesterone secretion in dairy cattle

The objective of this study was to investigate the effects of oxytocin infusion on corpus luteum (CL) function during early to mid-diestrus by measuring luteal size (LS) and luteal blood flow (LBF) along with plasma levels of progesterone (P4) and prostaglandin metabolites (13,14-dihydro-15-keto-prostaglandin F_2α, PGFM). On day (D) 7 of the estrus cycle (D1 = ovulation), seven cows received 100 IU of oxytocin (OXY) or placebo (PL) following a Latin square design. LS and LBF increased in both groups over time and no differences were observed between the groups. PGFM did not differ either within the groups over time or between the groups at any time point. P4 of the OXY group was higher compared to that of the the PL group 360 min after the infusion (p = 0.01) and tended to be higher at the time points 450 min, 48 h, and 72 h (all p = 0.08). Results from this study support the hypothesis that OXY is not directly involved in the mechanism(s) governing blood flow of the CL and has no remarkable effects either on luteal size or P4 and PGFM plasma levels. Further investigation is needed to elucidate the role of OXY in CL blood flow during early and late luteal phases.     

The Relationship between Linear Type Traits and Fertility Traits in High‐yielding Dairy Goats

Reproductive data collected from 13 medium‐size goat farms in central Mexico (582 goats), were used to investigate if linear type traits influenced reproductive performance of high‐yielding dairy goats. Data were analysed using multiple stepwise forward logistic regressions. Goats with the higher scores for strength were 1.43 times more likely (p < 0.05) to have larger litters (1.49 vs 1.39) than goats with lower strength. Goats approaching level rump angles were 1.68 times more likely (p < 0.01) to have larger litters, compared with goats with extremely sloped rump angles (1.48 vs 1.37). Goats with rear legs nearly straight (posty) in hock tended (p < 0.08) to presented larger litters than goats with rear legs slightly angled (sickled) in hock (1.49 vs 1.38). Less pronounced rump angle was favourable associated (p < 0.05) with litter weight (4.88 vs 4.53 kg) compared with goats with low pin bones. Goats with poorer udder texture were 1.42 more likely (p < 0.05) to have heavier litters than goats with good udder texture. Goats with faulty mammary system tended (p < 0.07) to have heavier litters than goats with good mammary system. Increasing udder height was associated (p < 0.05) with increasing number of services per conception. Goats with good fore udder attachment and non‐refined heads were at reduced risk (p < 0.05) for stillbirths. Goats with stronger median ligament were 65% less likely (p < 0.05) to lose their foetuses than those with looser udder support. Our findings indicate that a greater litter size and litter weight is expected in goats as rump angle becomes less sloped. This study showed that a relationship exists between strength of goats and litter size, and that some traits linked to high milk yield were antagonist to some reproductive traits.     

饲粮中主要谷物与谷物颗粒大小对早期罗曼褐壳蛋鸡生产性能和蛋品质的影响

试验旨在研究饲粮中主要谷物和谷物的几何平均粒径（GMD）对20～48周龄罗曼褐壳蛋鸡生产性能与蛋品质的影响。试验选择960只罗曼褐壳蛋鸡,采用完全随机设计为6×2×3,共分6个处理组．谷物饲粮分别为玉米和小麦,谷物粉碎粒度分别为6、8、10mm。每个处理设8个重复,每个重复20只。所有的饲粮配方设计中包含50％的玉米或小麦。每4周记录生产性能结果,第48周龄时测量蛋品质。试验结果表明,GMD为10mm时鸡采食量显著高于GMD为6和8mm时（P〈0.05）,说明粗糙的谷物粉碎粒度一定程度上有助于提高蛋鸡的采食量,而对蛋品质参数均没有显著影响。可以得出结论,谷物类型与谷物GMD除了影响早期罗曼褐壳蛋鸡采食量随谷物粉碎粒度的增加而增加外,其生产性能和蛋品质指标变化均不显著。     

Inhibitory action of Cerrado plants against mammalian and insect α-amylases

A total of 185 hexanic, dichloromethanic, ethanolic and hydroethanolic extracts from 24 species of Cerrado plants, were tested against Zabrotes subfasciatus, Acanthoscelides obtectus, and human saliva α-amylases. Twelve crude extracts presented inhibition rates greater than 80% against digestive α-amylases of the insect pest Z. subfasciatus, at a concentration of 1 mg mL⁻¹. These extracts were also tested against A. obtectus and human saliva α-amylases to verify their affinity and specificity of action. The hydroethanolic Kielmeyera coriacea stem bark extract presented a strong inhibitory potential, with IC₅₀ values of 110 μg mL⁻¹ for Z. subfasciatus and 272.12 μg mL⁻¹ for A. obtectus, in addition to a 97.09% reduction in enzyme activity of human saliva α-amylases at 125 μg mL⁻¹. The hexanic Aspidosperma macrocarpon root wood extract totally inhibited the activity of Z. subfasciatus α-amylases, reduced the enzyme activity of A. obtectus by 14.69% at 1 mg mL⁻¹, but did not alter the activity of human saliva α-amylases, thus characterizing greater inhibition affinity and specificity. The results suggest that the application of plant extracts against insect α-amylases represent a promising biotechnological tool for development of new insect pest control strategies, with noticeable affinity and specificity of action against different target enzymes.     

Mineral dialyzability in milk and fermented dairy products fortified with FeNaEDTA

Iron, zinc, and calcium dialyzability and ascorbic acid (AA) concentrations were evaluated in milk and yogurt fortified with FeNaEDTA (FE) or ferrous sulfate (FS) as a control, with or without AA addition. The values obtained for FE iron dialyzability in milk were much higher than those obtained for FS. The addition of AA to milk improved Fe dialyzability when using FS and slightly decreased Fe dialyzability in the FE-fortified nonfermented samples. Milk fermentation increased iron availability from both iron sources. Zinc and calcium dialyzability in products containing any of the two iron sources was increased in fermented milks. EDTA improved Zn dialyzability from intrinsic zinc in every manufactured dairy product. Whereas for milks fortified with FS and stored at 4 degrees C for 24 h, the AA content remained close to the original concentration, a higher AA degradation was observed when milks were fortified with FE.