Microencapsulation, chemical characterization, and antimicrobial activity of Mexican (Lippia graveolens H.B.K.) and European (Origanum vulgare L.) oregano essential oils.

The effect of solvent polarity (methanol and pentane) on the chemical composition of hydrodistilled essential oils (EO's) of Lippia graveolens H.B.K. (MXO) and Origanum vulgare L. (EUO) was studied by GC-MS. Composition of modified starch microencapsulated EO's was conducted by headspace-solid-phase microextraction (HS-SPME). The antimicrobial activity of free and microencapsulated EO's was evaluated. They were tested against Salmonella sp., Brochothrix thermosphacta, Pseudomonas fragi, Lactobacillus plantarum, and Micrococcus luteus. Thymol and carvacrol were among the main components of EO's and their free and microencapsulated inhibitory activity was tested against M. luteus, showing an additive combined effect. Chemical composition of EO's varied according to the solvent used for GC analysis and to volatile fraction as evaluated by HS-SPME. Thymol (both solvents) was the main component in essential oil of MXO, while carvacrol was the main component of the volatile fraction. EUO showed α-pinene (methanol) and γ-terpinene (pentane) as major constituents, the latter being the main component of the volatile fraction. EO's showed good stability after 3 months storage at 4°C, where antimicrobial activity of microencapsulated EO's remained the same, while free EO's decreased 41% (MXO) and 67% (EUO) from initial activity. Microencapsulation retains most antimicrobial activity and improves stability of EO's from oregano.

Thermal Balance in Male Water Buffaloes Transported by Long and Short Journeys.

Transport is a stressor that can cause physiological and metabolic imbalances in livestock, resulting in stress-induced hyperthermia. In water buffaloes, studies regarding the thermal state of animals during mobilization are scarce. Therefore, this study aimed to compare the thermal response of 1516 water buffaloes using infrared thermography (IRT) during 15 short trips (783 animals, 60,291 records, average duration = 50.33 min ± 5.48 min) and 14 long trips (733 animals, 56,441 records, average duration = 13.31 h ± 47.32 min). The surface temperature was assessed in 11 regions (periocular, lacrimal caruncle, nasal, lower eyelid, auricular, frontal-parietal, pelvic limb, torso, abdominal, lumbar, and thoracic) during seven phases from pasture to post-transport. It was found that the surface temperature of the periocular, lacrimal caruncle, nasal, auricular, frontal-parietal, pelvic limb, torso, abdominal, lumbar, and thoracic regions was significantly higher during SJs (+3 °C) when compared to LJs (p < 0.0001). In particular, the frontal-parietal region had a significant increase of 10 °C during the post-transport phase (p < 0.0001) in both groups, recording the highest temperatures during this phase. Likewise, a strong positive significant correlation between the different regions was found (r = 0.90, p < 0.0001). It is worth mentioning that the herding, loading, pre-, and post-transport phases were the ones where the greatest thermal response was recorded, possibly due to the influence of human interaction. Finally, a strong positive correlation (r above 0.9, p > 0.001) between the periocular, lacrimal caruncle, pinna, and pelvic limb was found. According to the results, SJ could be considered a stressful event that hinders thermal generation, contrarily to LJ.

Assessment of thermal changes in water buffalo mobilized from the paddock and transported by short journeys.

Evaluating the welfare of buffaloes during transport is key to obtaining and commercializing high-quality meat products; however, effective assessments require recognizing several stressors that activate physiological mechanisms that can have repercussions on the health and productive performance of species. The aim of this study was to evaluate the surface temperatures of different body and head regions in this species during events prior, and posterior, to transport for short periods; that is, from paddock to loading. The second goal was to determine the level of correlation between thermal windows. This study used infrared thermography (IRT) to evaluate the surface temperature of 624 water buffaloes (Buffalypso breed) during 12 short trips (average duration = 2 h ± 20 min) by focusing on 11 regions of the body (Regio corporis), in the head regions (Regiones capitis) the face regions (Regiones faciei), Orbital region (Regio orbitalis) with special attention to structures such as the lacrimal caruncle, periocular area and lower eyelid (Regio palpebralis inferior); nasal region (Regio nasalis) with special attention to nostril thermal window; and regions of the skull (Regiones cranii) such as auricular region (Regio auricularis) with special attention to auditory canal and frontal-parietal region (Regio frontalis-parietalis) and trunk region (Truncus regionis) such as thoracic and abdominal regions, regions of the vertebral column (Columna vertebralis) with the thoracic vertebral region (Regio vertebralis thoracis) and lumbar region (Regio lumbalis); and regions of the pelvis limb (Regiones membri pelvini). Recordings were made during seven phases: paddock (P1), herding (P2), corral (P3), chute handling (P4), shipping (P5), pre- (P6), and post-transport (P7). A total of 48,048 readings were obtained from 11 thermal windows. The results showed that the surface temperatures of the windows increased by as much as 5°C during P2, P3, P5, P6, and P7 compared to P1 and P4 (p < 0.0001). Differences of at least 1°C were also observed between thermal windows in the craniofacial, lateral corporal, and peripheral zones (p < 0.0001). Finally, a strong positive correlation (r = 0.9, p < 0.0001) was found between the thermal windows. These findings lead to the conclusion that the surface temperature of the craniofacial and corporal regions of buffaloes transported for short periods varied in relation to the phase of mobilization (from paddock to post-transport), likely as a response to stressful factors, since herding and loading increased the thermal values in each window. The second conclusion is that there are strong positive correlations between central and peripheral thermal windows.

Physiological responses of pigs to preslaughter handling: infrared and thermal imaging applications.

Loading, transport, unloading, lairage and stunning are the principle ante-mortem events that generate negative responses associated with stress in pigs. For this reason, it is important to verify the condition of animals throughout the supply chain in order to ensure their welfare and obtain, at the end of the slaughtering process, high-quality carcases and meat. Several studies have identified the indicators and samples that need to be taken into account to properly measure and evaluate the responses that these animals emit to the stressors involved. However, these procedures must be carried out quickly and by non-invasive means so as not to impede the flow of animals through the operations of loading, transport, unloading and lairage. Therefore, the objective of this review article is to analyse the stressful events that pigs experience during these events and discuss the use of the infrared thermography (IRT) as an alternative tool for measuring stress based on temperature changes on the surface of pigs' skin. We argue that infrared thermography can be used as a strategy to improve animal welfare during loading, transport, unloading and lairage by preventing fatigue and deaths, and decreasing negative impacts on meat quality, such as pale, soft and exudative (PSE), or dark, firm and dry (DFD) conditions.

Antioxidant Capacity and Cytotoxic Effects of Catechins and Resveratrol Oligomers Produced by Enzymatic Oxidation against T24 Human Urinary Bladder Cancer Cells.

In this work the polymerization of catechin, epicatechin, and resveratrol was carried out through a peroxidase oxidation process in order to improve the biological activity of these phenolic compounds. The antioxidant activity of the oligomers was evaluated by their ability to scavenge reactive oxygen species (ROS) and their capacity to chelate metal ions Fe2+ and Cu2+. The antitumor effect of the oligomers was determined by their ability to induce toxicity in the T24 human bladder cancer cell line. By enzymatic peroxidase oxidation, it was possible to produce oligomers of catechin, epicatechin, and resveratrol with antioxidant capacity significantly higher than their preceding monomers. The ROS scavenging capacity of the oligomers was 20 times higher than that of the monomers, while the ability of the oligomers to chelate metal ions increased up to about 1000 times. Our data show the antitumor effect of the oligomers of catechin, epicatechin, and resveratrol in the T24 cell line, which was similar to that observed with cisplatin. Oligomers of catechin, epicatechin, and resveratrol have great potential to be used as therapeutic agents for the treatment of oxidative stress-related diseases and bladder cancer.

Chitin extraction from Allopetrolisthes punctatus crab using lactic fermentation.

Chitin extraction from Allopetrolisthes punctatus, a crab species proliferating in Chile and Peru seashores, was carried out applying preliminary lactic ensilation. For this purpose, Lactobacillus plantarum sp. 47 isolated from Coho salmon was inoculated in crab biomass. Previously, fermentation parameters (carbon source, inoculum concentration and incubation temperature) to obtain peak lactic acid production and bacterial growth were studied. The optimal fermentation conditions were 10% inoculum, 15% sucrose and 85% crab biomass, producing 17 mg lactic acid/ g silage. Extracted and purified chitin, after 60 h fermentation, showed 99.6 and 95.3% demineralization and deproteinization, respectively, using low concentrated acids and bases. As a means of comparison, chitin was also extracted by chemical hydrolysis using high concentrated acids and bases, giving a lower yield and lower quality product.

Physiological responses in horses, donkeys and mules sold at livestock markets.

The horse welfare at markets includes fatigue, fear, fasting, dehydration and injuries. However, the scientific literature contains no readily-available information on the physiology responses of equids sold in livestock markets. The objective of the present study was to evaluate the effect of holding in livestock markets on gas exchange, the acid-base balance, energy metabolism, and the mineral and water balance in 4 types of horses (Equus caballus), donkeys (Equus asinus) and mules ((Equus asinus × Equus caballus). To this end, a total of 1,438 equine were utilized, after classification into six groups, as follows: mules, donkeys, culled horses, charrería horses (Mexican rodeo horses), foals, and ponies. The time intervals evaluated were 1 and 5 h. During the study, the animals received only water ad libitum. The physiological blood indicators assessed were: PvCO2 (mm Hg) and PvO2 (mm Hg) pressure, pH, glucose (mg/dL), lactate (mg/dL), bicarbonate levels (mmol/L), hematocrit (%), Na+, K+ and Ca2+ (mmol/L). The animals that presented the most marked physiological alterations were the mules and donkeys. The mules that remained in the corrals for 1 h presented increases (P < 0.0001) in the values for pO2, glucose, hematocrit and blood pH, compared to reference values (RV), while the donkeys had increases (P < 0.0001) in glucose and hematocrit values. By the end of the first hour at the exhibition corrals the equine were dehydrated, suffered fatigue, and suffered metabolic and compensatory problems, probably due to the combined effects of a series of factors that include transport and inadequate handling.

Effect of the number of incisions and use of local anesthesia on the physiological indicators of surgically-castrated piglets.

The objective of this study was to determine the effect of local anesthesia and the number of incisions performed on the physiological blood profile of piglets after surgical castration. A total of 60 male piglets were divided into five groups of 12 each, based on the surgical method employed and the use, or not, of local anesthesia, as follows: surgical castration using one horizontal incision in both testicles with (C1+L) and without (C1) local anesthesia; surgical castration using two vertical scrotal incisions with and without local anesthesia (C2+L and C2); and control piglets which were removed from their pens and held head-down by their hind limbs for approximately 90 s to simulate castration (SIM). Reference blood samples were drawn 24 h before castration (RV), immediately after surgery or simulated castration (PC), and at 24 and 48 h post-castration, to determine physiological profiles including; pH, hematocrit, glucose, electrolytes, lactate, pCO2 (mmHg), SO2 (mmHg), and bicarbonate. Results showed increases in lactate and hematocrit immediately after surgical or simulated castration with decreases in pH, HCO3- and base excess (BE). Surgical castration produced marked alterations of the physiological profile, detected by reduced pH and HCO3, higher lactate levels and BE alterations. These changes indicated metabolic acidosis that was greater in the piglets castrated surgically with one horizontal incision than in those castrated with two vertical incisions. More research is needed on the use of lidocaine during surgical castration, as it showed no effect on physiological profile in this study, but did alter hematocrit values.

Effect of Nanoemulsified and Microencapsulated Mexican Oregano (Lippia graveolens Kunth) Essential Oil Coatings on Quality of Fresh Pork Meat.

Fresh meat is a highly perishable food. This work aimed to evaluate the influence of Mexican oregano (Lippia graveolens Kunth) incorporated into active coatings (ACs) spread on fresh pork meat as free (FEO), nanoemulsified (NEO), and microencapsulated (MEO) essential oil (EO), on its microbiological, physicochemical and sensory properties during 15 d at 4 ± 1 °C. Thymol and γ-terpinene were identified in the EO. In vitro effect of 2.85 mg EO/cm2 was tested against Brochothrix thermosphacta, Micrococcus luteus, Lactobacillus plantarum, Pseudomonas fragi, and Salmonella Infantis. FEO antioxidant capacity (DPPH assay) was significantly higher than that of thymol, NEO and MEO (93.53%, 89.92%, 77.79%, and 78.50% inhibition, respectively), and similar to BHA (96.03%) and gallic acid (95.57%). FEO, NEO, and MEO ACs on meat caused growth inhibition of lactic acid bacteria (5 log population reduction) and Pseudomonas spp. (4 log reduction), whereas ≤1.5 log population reduction was observed for B. thermosphacta and Salmonella Infantis. Meat microbiota was more efficiently controlled by MEO than by FEO or NEO. ACs delayed lipid and oxymyoglobin oxidation of fresh pork meat. After 15 d of cold storage meat added with EO coatings was desirable for panelists, whereas untreated (UT) samples were undesirable. Active coatings are a significant alternative method for fresh meat preservation.

In vivo analysis of effects of venom from the jellyfish Chrysaora sp. in zebrafish (Danio rerio).

The jellyfishes of the genus Chrysaora are present in all of the world's oceans, but the toxicity of their venoms has not yet been thoroughly characterized. The zebrafish as a toxicology model can be used for general toxicity testing of drugs and the investigation of toxicological mechanisms. The aim of this study was to evaluate the effect of crude venom from jellyfish Chrysaora sp., a species of jellyfish observed in the tropical lagoons of the Gulf of Mexico, on the zebrafish Danio rerio. Juvenile zebrafish were injected with different concentrations of venom from Chrysaora sp. via intraperitoneal and subcutaneous injections. The effects of the venom were determined by histopathological analysis and through the measurement of hemolytic and phospholipase A2 activities. The crude venom was examined by SDS-PAGE. The effect of sublethal concentrations of crude venom from Chrysaora sp. on D. rerio was hemorrhaging in the eyes, while the histopathological analysis demonstrated that the primary organs targeted were the pseudobranch, which displayed hyperemia, and the gill, which displayed hyperplasia and hypertrophy. The blood analysis exhibited hemolysis, nuclear abnormalities, and echinocytes by the action of phospholipase A2, which was determined to have 596 units of activity/mg of protein in the venom. The crude venom has proteins with molecular weights ranging from 250 to 6 kDa, with more density in the bands corresponding to 70, 20 and 15 kDa. The venom of Chysaora sp. caused disturbances in circulation associated with vascular dilation due to the localized release of inflammatory mediators. The hemolysis of erythrocytes was caused by the action of phospholipase A2. These findings not only provide an excellent study model but also have a great pharmacological potential for designing new drugs and for the elucidation of the mechanisms of action of and treatment against stings.

Adaptation of ostriches to transport-induced stress: physiometabolic response.

This study assessed the effects of five different transport periods on physiometabolic responses and gas exchange in ostriches. It included 138 ostriches that were assigned to five experimental groups. G1 included 78 birds that were set aside as a reference group (RG). Each one of the four remaining experimental groups included 20 ostriches, which were transported in the following manner: G1 on one occasion for a period of 1 h; G2 on one occasion for 2 h; and G3 on one occasion for 3 h; G4a was made up of the ostriches from G1 but they were shipped on a second occasion, for 2 h (making a total of 3 h); finally, G4b was that same group, but after a third transport period, on this occasion during 3 h (for a total of 6 h). Groups G2 and G3 presented the most marked blood alterations (P < 0.05), including an increase in pH, hypocapnia (27.8 ± 0.80 mmHg), hypernatremia (171.75 ± 1.84 mmol/L), hypocalcemia (0.95 ± 0.03 mmol/L), and hyperglycemia (224.05 ± 3.94 mg/dL). Also, group G3 presented the lowest hematocrit values (26.5 ± 0.47 HTC%). Therefore, transporting ostriches with no prior experience for 3 h caused the most pronounced physiometabolic changes.

Actividad enzimática de proteasas de Cyprinus carpio (Cypriniformes: Cyprinidae) extraídas de una laguna contaminada en México / Enzymatic activity of proteases from Cyprinus carpio (Cypriniformes: Cyprinidae) captured in a polluted lagoon in Mexico

ResumenLa carpa común (Cyprinus carpio) es un organismo acuático de valor comercial que puede sobrevivir en ambientes contaminados; la carpa contiene enzimas proteolíticas de importancia fisiológica y potencial aplicación industrial. El objetivo de este estudio fue purificar parcialmente y determinar la actividad proteolítica a diferente pH de proteasas de carpas colonizando un ambiente contaminado. Se capturaron tres carpas en diferentes zonas de la laguna contaminada de Zumpango (México) a 1 m de profundidad máxima. El extracto crudo se obtuvo del músculo dorsal mediante extracción acuosa y se fraccionó con (NH4)2SO4 saturado a 20 %, 50 % y 80 %. Posteriormente, se seleccionaron las fracciones obtenidas con (NH4)2SO4 saturado a 50 % y 80 % por su alta actividad proteolítica, se concentraron por ultrafiltración con membranas de corte de peso molecular de 100 kDa, y se analizaron por electroforesis en geles de poliacrilamida con dodecil sulfato de sodio (SDS-PAGE). La actividad proteolítica del extracto crudo fue significativamente mayor (19.7-20.3 U / mg) a pH 2, 5, y 7 (P < 0.001). Las fracciones obtenidas con (NH4)2SO4 saturado a 20 %, 50 % y 80 % presentaron actividades proteolíticas óptimas a pH 5 (2.8 U / mg) y pH 6 (2.2 U / mg); pH 6 (4.3 U / mg) y pH 3-4 (3.6 - 3.7 U / mg); pH 3 (10.8 U / mg) y pH 10 (10.6 U / mg); respectivamente. Las subfracciones con peso molecular < 100 kDa obtenidas con (NH4)2SO4 saturado a 50 % y 80 % tuvieron máxima actividad proteolítica a pH alcalino. La subfracción < 100 kDa, obtenida con (NH4)2SO4 saturado a 80 % tuvo la mayor actividad proteolítica (37.3-43.7 U / mg) a pH 8-10, factor de purificación de 3 y 19.1 % de recuperación. Trece proteínas entre 9.8 a 104.8 kDa se identificaron en el extracto proteolítico crudo. Las proteínas de 31 - 33 y 39 - 41 kDa tuvieron la concentración más alta en las fracciones estudiadas, sugiriendo la posible predominancia de serín y aspartil proteasas, respectivamente. Nosotros sugerimos que la presencia de proteasas con máxima actividad a pH alcalino está relacionada con la adaptación de C. carpio a aguas contaminadas con pH alto. Aunque estos peces son inadecuados para el consumo humano, pueden ser empleados como materia prima para la producción de proteasas destinadas para varias industrias, incluido el tratamiento de aguas residuales.

AbstractCommon carp (Cyprinus carpio) is an aquatic organism of commercial value able to survive in polluted environments; carps contain proteolytic enzymes of physiological importance and potential industrial application. The objective of this work was partially purify and study the proteolytic activity at different pH of carp proteases living in a polluted environment. Three carps were captured in different zones of Zumpango polluted lagoon (Mexico) at 1 m of maximum deep. Protease crude extracts were obtained from dorsal muscle by aqueous extraction and fractionated by 20 %, 50 %, 80 %-saturated (NH4)2SO4. Fractions extracted with 50 % and 80 %-saturated (NH4)2SO4 were selected for their high proteolytic activity and concentrated by ultrafiltration through 100 kDa molecular weight cutoff membranes and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The crude proteolytic extract had significantly higher activity (19.7 - 20.3 U / mg) at pH 2, 5, and 7 (P < 0.001). Fractions obtained with 20 %, 50 % and 80 % - saturated (NH4)2SO4 showed peak activity at pH 5 (2.8 U / mg) and pH 6 (2.2 U / mg); pH 6 (4.3 U / mg) and pH 3 - 4 (3.6 - 3.7 U / mg); pH 3 (10.8 U / mg) and pH 10 (10.6 U / mg); respectively. Subfractions of < 100 kDa, obtained with 50 % and 80 %-saturated (NH4)2SO4, had peak proteolytic activity at alkaline pH. A < 100 kDa fraction, obtained with 80 %-saturated (NH4)2SO4, had the highest proteolytic activity (37.3 - 43.7 U / mg) at pH 8 - 10, purification factor of 3 and 19.1 % recovery. Thirteen proteins between 9.8 to 104.8 kDa were identified in the crude extract. Peak protein concentration was observed for 31 - 33 and 39 - 41 kDa, suggesting the possibility predominance of serine- and aspartyl- proteases, respectively. We suggest this protease with maximum activity at alkaline pH is related to the adaptation of C. carpio to polluted waters with high pH. Although unsuitable for human consumption, these organisms can be a source of protease production aimed to several uses as in the industry and waste water treatment among others.

Stability studies on astaxanthin extracted from fermented shrimp byproducts.

To the best of our knowledge, stability studies on astaxanthin contained in carotenoproteins extracted from lactic acid fermented shrimp byproduct have never been reported. Carotenoprotein powder, containing 1% free astaxanthin, was subjected to oxidation factors of illumination, oxygen availability, and temperature, using synthetic astaxanthin as a control. The individual effects as well as first and second degree interactions were studied on natural and synthetic free astaxanthin stability. Air and full light were the two individual factors with the highest effects on astaxanthin oxidation. Sixty-two and 46% natural and synthetic astaxanthin, respectively, oxidized when exposed to air for 8 weeks of storage, whereas 35 and 28% of natural and synthetic astaxanthin, respectively, oxidized under full light. Ninety-seven and 88% of natural and synthetic astaxanthin, respectively, oxidized under a combination of full light, air, and 45 degrees C at 8 weeks of storage. Storage in the dark, nonoxygen, and 25 degrees C were the treatments that efficiently minimized astaxanthin oxidation. Natural astaxanthin from fermented shrimp byproduct presented moderate stability levels. Although natural astaxanthin oxidized faster than the synthetic pigment, its stability may improve by antioxidant and polymer addition.

Aspectos relevantes del bienestar del cerdo en tránsito / Relevant aspects of swine welfare in transit

Current globalization policies demand animal welfare standards on animal transportation. In spite of international tendencies to commercialize meat cuts while decreasing live animal transit, transport is still one of the major problems in terms of animal welfare, besides carcass and meat by-products' quality. The present review analyzes, in general terms, the different definitions on animal welfare and factors affecting pig welfare in transit. Several case studies are referred to as examples, showing the animal response to stress during transport and its effects on both meat quality and the economic impact. In addition, legal requirements and Mexican regulations for pig transportation are also described. Information in regard to swine stress and welfare in transit, considers a number of factors that alter the animal metabolic homeostasis with subsequent negative effects on pork quality. It is conclude that knowledge on basic animal behavioral and physiological needs during transport, as well as a suitable training of personnel, are necessary for reducing animal welfare problems. Last but not least, some recommendations on handling practices are given in order to improve swine welfare during transit.

En el marco actual de la globalización, hablar de transporte animal conlleva realizar prácticas que mejoren el bienestar animal, aunque existe la tendencia mundial de incrementar el mercado de la carne en cortes, así como disminuir el comercio de animales vivos, el transporte constituye uno de los factores más preocupantes en términos del bienestar animal, además del impacto en la calidad de la canal y los subproductos respectivos. En la presente revisión se analizan, en términos generales, las distintas definiciones de bienestar animal, así como los factores más importantes que alteran el bienestar de los cerdos durante el transporte; se describen los requisitos legales y la normatividad mexicana para el transporte de animales, así como varios estudios de la respuesta fisiológica del animal durante condiciones adversas del transporte, se señalan, además, las repercusiones sobre la calidad de la carne y su impacto económico. La información con respecto al estrés de los cerdos y su bienestar durante el transporte, considera numerosos factores que alteran el equilibrio homeostático animal y que propician efectos negativos sobre calidad de la carne. Se concluye que tanto el conocimiento de la biología de la especie, como un entrenamiento del personal, son necesarios para disminuir los problemas de bienestar. Finalmente, se recomiendan algunas prácticas derivadas de los hallazgos presentados, con el fin de que se mejore el bienestar de los cerdos en tránsito.