Factors affecting N-acetyl-ß-D-glucosaminidase as an indicator for mastitis detection in dairy sheep.

The study of new indirect methods for mastitis detection is of great relevance both at the economic level of the farm and dairies, and in terms of consumer health, and animal welfare. These methods help us to monitor the disease and speed up the decision-making process on treatment of the affected animal and the destination of the milk. The main aim of this work was to study the effect of intramammary infection and other non-infectious factors on the activity of the enzyme N-acetyl-ß-D-glucosaminidase (NAGase) in milk, in order to evaluate its use as an indicator for the early diagnosis of mastitis in sheep that could be less expensive, easier to measure and a better marker of inflammation or complementary to existing methods such as somatic cell count (SCC). Seven biweekly samplings were carried out, in which NAGase activity, SCC and milk were analyzed. Glands were classified according to their sanitary status based on the results of the SCC and bacteriological analysis. Non-infectious factors such as lactation stage, parity number and milking session had a statistically significant effect on NAGase values, finding the highest NAGase values at the onset and end of the study, in infectious mastitic glands of multiparous females and at morning milking. However, among the NAGase variation factors studied, the health status of the gland was the factor that caused the highest variation in enzyme levels, with infectious mastitic glands showing higher values than healthy glands. The predictive ability of NAGase was also studied by means of several logistic regression models, with the one that included NAGase together with lactation stage and parity obtaining the best results if sensitivity is to be prioritized, or the model that included NAGase, lactation stage, parity, milking and production if specificity is to be prioritized. From the results obtained, it can be concluded that the use of NAGase as an intramammary infection detection method in sheep can be useful when non-infectious factors that cause changes in the concentration of the enzyme are also considered.

Functional immune boosters; the herb or its dead microbiome? Antigenic TLR4 agonist MAMPs found in 65 medicinal roots and algae's.

Background: Humans have been consuming medicinal plants (as herbs/ spices) to combat illness for centuries while ascribing beneficial effects predominantly to the plant/phytochemical constituents, without recognizing the power of obligatory resident microorganism' communities (MOCs) (live/dead bacteria, fungus, yeast, molds etc.) which remain after industrial microbial reduction methods. Very little is known about the taxonomic identity of residual antigenic microbial associated molecular patterns (MAMPs) debris in our botanical over the counter (OTC) products, which if present would be recognized as foreign (non-self) antigenic matter by host pattern recognition receptors (PRRs) provoking a host immune response; this the basis of vaccine adjuvants. As of today, only few research groups have removed the herbal MAMP biomass from herbs, all suggesting that immune activation may not be from the plant but rather its microbial biomass; a hypothesis we corroborate. Purpose: The purpose of this work was to conduct a high through put screening (HTPS) of over 2500 natural plants, OTC botanical supplements and phytochemicals to elucidate those with pro-inflammatory; toll like receptor 4 (TLR4) activating properties in macrophages. Study Design: The HTPS was conducted on RAW 264.7 cells vs. lipopolysaccharide (LPS) E. coli 0111:B4, testing iNOS / nitric oxide production (NO2-) as a perimeter endpoint. The data show not a single drug/chemical/ phytochemical and approximately 98 % of botanicals to be immune idle (not effective) with only 65 pro-inflammatory (hits) in a potency range of LPS. Method validation studies eliminated the possibility of false artifact or contamination, and results were cross verified through multiple vendors/ manufacturers/lot numbers by botanical species. Lead botanicals were evaluated for plant concentration of LPS, 1,3:1,6-ß-glucan, 1,3:1,4-ß-D-glucan and α-glucans; where the former paralleled strength in vitro. LPS was then removed from plants using high-capacity endotoxin poly lysine columns, where bioactivity of LPS null "plant" extracts were lost. The stability of E.Coli 0111:B4 in an acid stomach mimetic model was confirmed. Last, we conducted a reverse culture on aerobic plate counts (APCs) from select hits, with subsequent isolation of gram-negative bacteria (MacConkey agar). Cultures were 1) heat destroyed (retested/ confirming bioactivity) and 2) subject to taxonomical identification by genetic sequencing 18S, ITS1, 5.8 s, ITS2 28S, and 16S. Conclusion: The data show significant gram negative MAMP biomass dominance in A) roots (e.g. echinacea, yucca, burdock, stinging nettle, sarsaparilla, hydrangea, poke, madder, calamus, rhaponticum, pleurisy, aconite etc.) and B) oceanic plants / algae's (e.g. bladderwrack, chlorella, spirulina, kelp, and "OTC Seamoss-blends" (irish moss, bladderwrack, burdock root etc), as well as other random herbs (eg. corn silk, cleavers, watercress, cardamom seed, tribulus, duckweed, puffball, hordeum and pollen). The results show a dominance of gram negative microbes (e.g. Klebsilla aerogenes, Pantoae agglomerans, Cronobacter sakazakii), fungus (Glomeracaea, Ascomycota, Irpex lacteus, Aureobasidium pullulans, Fibroporia albicans, Chlorociboria clavula, Aspergillus_sp JUC-2), with black walnut hull, echinacea and burdock root also containing gram positive microbial strains (Fontibacillus, Paenibacillus, Enterococcus gallinarum, Bromate-reducing bacterium B6 and various strains of Clostridium). Conclusion: This work brings attention to the existence of a functional immune bioactive herbal microbiome, independent from the plant. There is need to further this avenue of research, which should be carried out with consideration as to both positive or negative consequences arising from daily consumption of botanicals highly laden with bioactive MAMPS.

An efficient analysis of oblique reflection of airborne ultrasound beams from thin membranes for gas sensing.

The use of the angular spectrum method (ASM) to simulate the reflection of airborne ultrasound beams from a thin membrane separating air from a mixture of air and another gas is examined. The main advantage of this method is its high computing speed and efficiency for practical design calculations, suitable for sensing applications. The implemented ASM code is validated against custom Rayleigh integral code in a pure propagation simulation. In addition, ultrasound beam reflection calculations using ASM with finite element numerical results and experimental measurements are compared, finding good agreement in both cases. Then, ASM is used to estimate the sensitivity of specular reflection signals to variations in the composition of the incidence medium as a function of the angle of incidence. Conditions for which a reflection signal using inexpensive commercial ultrasound emitter/receiver at 40 kHz, in a simple configuration, offer a high enough sensitivity suitable for monitoring air quality indoors are found.

Foaming and Structural Studies on the Acidic Subunit of Amaranth 11S Globulin Modified with Antihypertensive Peptides as a Function of pH and Ionic Strength.

Some studies aimed at revealing the relationship between protein structure and their functional properties. However, the majority of these reports have been carried out using protein isolates. There are limited reports on the possible relationship between the functional properties and the structure of a purified protein. In this work the amaranth 11S globulin acidic subunit (AAC) and five mutations of the same protein that were modified in their variable regions with antihypertensive peptides (VYVYVYVY and RIPP), were analyzed at two ionic strength (2.9 and 17.6 g/L NaCl) and pH (3.0-7.0). Results revealed better solubility for the proteins mutated at the terminal ends (AACM.1 and AACM.4) and lower solubility for the protein inserted with RIPP peptide. Spectroscopy studies revealed an increase of ß-sheet structure at high salt concentration for all proteins. It was also observed that salt concentration acted as a modulator, which allowed a better foam features for all modified proteins limiting movement of side chains and reducing red-shifted displacement of λmax. All proteins showed foam capacity ranging from 76 to 93% although foam stability was twofold better for modified proteins than for AAC at high salt concentration. This study allowed better understanding about the structural changes that influence the foaming properties of engineered proteins.

Comparison of ELISA using recombinant LipL32 and sonicated antigen of leptospira for detecting bovine leptospirosis.

Leptospirosis is one of the most widely distributed zoonosis in the world. Bovine leptospirosis is a serious problem in bovine production, causing reproductive losses. The aim of this work was to compare recombinant LipL32 with sonicated antigen for detecting anti-Leptospira IgG antibodies in bovine serum using ELISA. The Microscopic Agglutination Test (MAT) is used as the gold standard. Sonicated antigen from cultures of Leptospira interrogans serogroup Icterohaemorrhagiae serovar copenhageni (strain M20) was used for the eELISA and rLipL32 for the rELISA. The performance of these assays was evaluated using serum samples from 166 bovines, 69 MAT positive and 97 MAT negative. At the optimal cut-off point recommended by the receiver operating characteristic (ROC) curve analysis, the sensitivity and specificity values were 98.6% and 97.9%, respectively, for eELISA, and 85.5% and 86.6% respectively, for rELISA. The value for the area under the ROC curve was 0.998 (0.994-1.0) (CI 95%) for eELISA and 0.929 (0.891-0.968) (CI 95%) for rELISA. The ROC curves for rLipL32 and sonicated antigen showed statistically significant differences (z = -3.826; p = 0.000). A three-way comparison showed statistically significant differences in the sensitivity and specificity of rELISA and eELISA. Our results showed that eELISA was more specific and sensitive than rELISA. The difference in performance (eELISA-rELISA) was 13.4% (4.03-23.28) (CI 95%) for sensitivity and 11.34 % (4.07-19.56) (CI 95%) for specificity. Our results show that the eELISA has a better diagnostic performance than rELISA for the detection of anti-Leptospira IgG antibodies in bovine serum.

Engineering Concanavalin B to Release Bioactive Peptides against Metabolic Syndrome.

Metabolic syndrome is a severe public health issue characterized by multiple metabolic disturbances. Current treatments prescribe a particular drug for each of them, producing multiple side effects. As a first step towards a more integral approach, we applied our recently described methodology to design single proteins, based in the Concanavalin B scaffold (1CNV), that contain several bioactive peptides (BPs), including antioxidant and lipid-lowering activities as well as inhibitors of dipeptidyl peptidase IV (DPPIV) and the angiotensin converting enzyme. Modified Concanavalin (CNV44), the designed protein that showed the best in silico properties, was expressed in high yields in E. coli and purified to homogeneity. After in vitro digestion with gastrointestinal enzymes, all the biological activities tested where higher in CNV44 when compared to the non-modified protein 1CNV, or to other previous reports. The results presented here represent the first in vitro evidence of a modified protein with the potential to treat metabolic syndrome and open the venue for the design of proteins to treat other non-communicable diseases.

Virulence of Metarhizium brunneum (Ascomycota: Hypocreales) Strains Against Stinkbugs Euschistus heros and Dichelops furcatus (Hemiptera: Pentatomidae).

Three strains of fungi belonging to the genus Metarhizium Sorokin (ARSEF 4556, ARSEF 3297, native strain) were assayed against adults and nymphs of the Neotropical brown stinkbug Euschistus heros (F.) and the green-belly stinkbug Dichelops furcatus (F.). The most virulent strain, ARSEF 4556, caused over 90% mortality. The average survival time of the second and fifth instar nymphs and adults following immersion in 1 × 108 conidia ml-1 was 4.8, 5.7, and 5.2 d, respectively. The second instar nymphs were more susceptible than the adults. The LC50 values and median survival times for second instar and adult E. heros were 1.6 × 107 and 3.1 × 107 conidia ml-1 and 6 and 8 d, respectively. Eggs of E. heros and the closely related stinkbug, D. furcatus, were highly susceptible to ARSEF 4556 with the mean mortality of eggs immersed in 1 × 108 conidia ml-1 being 77.4% and 89.7%, respectively. The strain 3297 showed also good aptitudes for stinkbugs control with mortalities higher than 80% against nymphs and adults and eggs mortalities of 75.5% for E. heros and 79.6% for D. furcatus. This study has shown that it is possible to have a two-pronged control strategy, targeting adults and to reduce oviposition and targeting egg clusters to prevent emergence and dispersal of nymphs. Besides early instars of nymphs have been shown to be more susceptible to the fungal strains than late instars and adults.

Bartonella in dogs and fleas from Tulancingo, Hidalgo, Mexico.

Bartonella sp. infection is quite common in free-roaming dogs in many tropical countries. However, limited information is available of the presence of these pathogens in Mexico. The present study looked at prevalence of Bartonella exposure and/or infection in dogs and their fleas in Central Mexico. Blood samples were collected from 31 stray dogs in August 2014 at the municipal pound, Tulancingo, Mexico, as well as fleas on 26 of them. Bartonella seropositivity was 46.9%, including 35.5% for Bartonella henselae, 45% for Bartonella clarridgeiae and 32.2% for Bartonella vinsonii subsp. berkhoffii. Three (9.7%) dogs were polymerase chain reaction (PCR) positive for the Bartonella gltA gene. Partial sequencing of that gene revealed that these three dogs were infected with B. henselae. In total, 86 fleas were collected from 26 dogs (range 1-9 fleas per dog), including 52 Ctenocephalides felis and 34 Ctenocephalides canis. Of 40 pools of fleas (20 pools of C. canis and 20 pools of C. felis), five (12.5%) were PCR positive for the Bartonella sp. gltA gene, including three C. canis pools (five fleas) and two C. felis pools (three fleas). All sequences showed 99.25% to 100% homology with B. henselae Houston I.

Soil drenching with entomopathogenic fungi for control of the soil-dwelling life stages and adults of the same generation of Spodoptera littoralis (Boisd.) (Lepidoptera: Noctuidae).

Four Beauveria bassiana and three Metarhizium brunneum isolates were evaluated, as soil drenches, against Spodoptera littoralis prepupae. Treatment efficacy was determined by assessing total mortality during development from prepupae through to pupae and adults; mortality and sub-lethal effects on reproduction were also quantified for adults emerging from surviving prepupae/pupae. All isolates were pathogenic but overall mortality varied between 31.7 and 83.3% (0% for control); average survival time was 7.5-10.5 days (14.0 days for control). From 1.7-15.0% of adults emerging from surviving prepupae/pupae were deformed (0% in control). Contact with fungal suspensions as prepupae/pupae caused a significant reduction in fecundity of emerging adult females (15-58.9%), and a significant reduction in egg viability (6.8-28.4%) compared with controls. Two isolates were selected for virulence evaluation against S. littoralis prepupae. The LC50s were 1.7 × 107 and 1.8 × 107 conidia ml-1 and the median survival times were 7 and 6 days for isolates EAMa 01/58-Su and EAMb 09/01-Su, respectively. Destruxin A was present in pupae developing from prepupae treated with isolates EAMa 01/58-Su (0.010 ± 0.002 µg pupae-1) and EAMb 09/01-Su (0.015 ± 0.003 µg pupae-1). The use of entomopathogenic fungi as soil drenches could be a key component of S. littoralis IPM strategies due to direct reductions in the number of soil-dwelling life stages and, also, the significant reduction in reproductive potential of surviving adults.

Assessment of artifacts reduction and denoising techniques in Electrocardiographic signals using Ensemble Average-based method.

BACKGROUND AND OBJECTIVE: Outpatient vital signs monitoring has a key role in medical diagnosis and treatment. However, ambulatory vital signs monitoring has great challenges to overcome, being the most important, the reduction of noise and Motion Artifacts, which hide essential information, particularly in Electrocardiographic signals. Despite efforts being made to reduce these artifacts, a comparative performance assessment of proposed techniques does not exist to the best of our knowledge and there are no enhancement level measurements obtained by the signals in the artifacts reduction. This article presents a new method based on Ensemble Average for the performance comparison of reported techniques for the processing and reduction of noise and artifacts in Electrocardiographic signals. METHODS: The comparison was done using a dataset composed by six synthetic noised Electrocardiographic signals and six real one acquired from healthy volunteers that intentionally introduced Motion Artifacts. Several techniques that have reported positive results in the enhancement of Electrocardiographic signals were applied to this dataset to compare their performance in the reduction of Motion Artifacts. The Signal-to-Noise Ratio and the Ensemble Average as a distortion measurement were used to compare the performance of algorithms to produce an enhanced signal. RESULTS: In agreement to previous reports, all studied methods show a significant improvement of the Signal-to-Noise Ratio. Concerning the distortion of the waveform, although all methods caused high distortion on the enhanced signal waveform, the Wavelet-ICA method showed the best performance. The percentage of signal distortion introduced by denoising techniques was evaluated through the proposed Ensemble Average Electrocardiographic method. CONCLUSIONS: It was found that the proposed method based on Ensemble Average offers a complementary way to measure the performance of denoising techniques when considering the introduced distortion in the waveform segments once the artifact reduction process was applied and not only the change in the Signal-to-Noise Ratio.

Late-onset sepsis and mortality among neonates in a Brazilian Intensive Care Unit: a cohort study and survival analysis.

A cohort study was performed from January 2014 to December 2016 in a Brazilian neonatal intensive care unit, including neonates with high risk for infection and death. We estimated bloodstream infection (BSI) incidence and conducted a survival analysis, considering the time to death and to the first episode of BSI as outcomes, comparing very low birth weight (VLBW) neonates with the remaining neonates. An extended Cox model was performed and the hazard ratio (HR) was calculated for different time periods. The study had 1560 neonates included, the incidence and the incidence density of BSI was 22% and 18.6 per 1000 central venous catheter-days, respectively. Considering VLBW neonates as the reference group, the HR for time to death was 4.06 (95% CI 2.75-6.00, P < 0.01) from day 0 to 60 and for time to the first episode of BSI was 1.76 (95% CI 1.31-2.36, P < 0.01) from day 0 to 36. Having the heavier neonates group as reference, the HR for time to the first episode of BSI was 2.94 (95% CI 1.92-4.34, P < 0.01) from day 37 to 90. Late-onset neonatal sepsis prevention measures should consider the differences in risk during time, according to neonates' birth weight.

Eficacia y seguridad de tres esquemas para la preparación del colon para colonoscopia (polietilenglicol (PEG) 4 litros (4 L) (dosis única) vs. PEG 4 L dividido (2 L + 2 L) vs. PEG 2 L dividido de volumen bajo (1 L + 1 L): ensayo clínico controlado aleatorizado / A randomized controlled clinical trial of the efficacy and safety of colonoscopy preparation using a single four liter dose of polyethylene glycol (PEG) vs. two 2 liter doses of PEG vs. two low volume (1L + 1L) doses of PEG

Resumen Introducción: la colonoscopia es el examen estándar de oro para evaluar la mucosa del colon. De la limpieza del colon en la preparación intestinal para colonoscopia depende el hallazgo de pólipos, que pueden ser adenomatosos con potencial maligno y con la posibilidad de degenerarse en cáncer de colon. Objetivo: comparar la eficacia y la seguridad de tres tipos de preparaciones para la limpieza del colon: dosis única de polietilenglicol (PEG) 4 litros (4 L) y dosis divididas: PEG 4 L dividido (2 L + 2 L) y PEG 2 L dividido (1 L + 1 L) de volumen bajo. Métodos: en pacientes con una colonoscopia electiva de una clínica universitaria, se realizó un ensayo clínico controlado aleatorizado y ciego (para el médico que evaluó la limpieza del colon). Se asignaron 74 pacientes para cada grupo. El parámetro principal de eficacia fue la preparación integral de calidad adecuada medida con la escala de Boston, y los parámetros secundarios fueron el porcentaje de eventos adversos, la tolerabilidad y la tasa de detección de pólipos. Resultados: la preparación completa de todo el colon fue significativamente de mayor la calidad en la alternativa de 4 L divididos (2 L + 2 L), seguida de la otra alternativa dividida (1 L + 1 L) y menor en la dosis única (79,7 %, 75,7 % y 63,5 %, respectivamente, p = 0,019); también se encontraron diferencias en la detección de pólipos (13,5 %, 24,3 % y 9,5 %, p = 0,037) y sin diferencias en la presentación de al menos un evento adverso (p = 0,254) ni en la tolerabilidad (p = 0,640). Conclusiones: las dos preparaciones de dosis dividida tienen una mayor eficacia en la limpieza del colon en comparación con la dosis única de 4 L y en la detección de pólipos, mientras que no se evidencian diferencias en las preparaciones para la ocurrencia de eventos adversos y la tolerabilidad. La dosis de PEG 2 L dividido puede ser una muy buena opción para las preparaciones de colonoscopia electiva.

Abstract Introduction: Colonoscopy is the gold standard for evaluation of the colonic mucosa. Colon cleansing in preparation for colonoscopy depends on finding of polyps which can be adenomatous with malignant potential and the possibility of degenerating into colon cancer. Objective: This study's objective was to compare the efficacy and safety of three types of preparations for colon cleansing: a single four liter dose of polyethylene glycol (PEG) vs. two 2 liter doses of PEG vs. two low volume (1L + 1L) doses of PEG. Methods: This is a randomized controlled clinical trial of patients who underwent elective colonoscopy at a University clinic. It was blinded for the doctor who evaluated colon cleansing. Seventy four patients 74 patients were randomized into each group. The main parameter of effectiveness was integral preparation of adequate quality measured on the Boston scale. Secondary parameters were the percentage of adverse events, tolerability and detection rate of polyps. Results: Complete preparation of the entire colon was achieved significantly more often with 4 liters divided into two 2 liter doses followed by the other divided alternative (1 L + 1 L). It was achieved least frequently with in the single dose: 79.7%, 75.7% and 63.5%, respectively, p = 0.019. Differences were also found in the detection of polyps (13.5%, 24.3% and 9.5%, respectively, p = 0.037). ) There were no differences in presentation of at least one adverse event (p = 0.254) or in tolerability (p = 0.640). Conclusions: The two divided dose preparations had higher colon cleansing and polyp detection efficacies than did the single 4L dose while there were no differences in occurrence of adverse events and tolerability. The divided PEG 2L dose could be a very good option for elective colonoscopy preparation.

Effect of the pulsation type (alternate or simultaneous) on milk yield and health status of the mammary gland of Murciano-Granadina goats.

Recent research into how the pulsation type affects goat milking concluded that alternate pulsation (AP) and simultaneous pulsation (SP) similarly affect the milking efficiency (milking duration and milking fractioning) and teat end condition after milking when both pulsations are employed in short-term experiments. However, the use of SP in high pipeline milking machines led to higher vacuum fluctuations, which, in the long term, may entail a potential risk factor for the mammary gland health status. The aim of this work was to study how AP and SP affect the milk yield, mammary gland health status, and milk composition of goats in the long term, during a complete lactation period, in a high pipeline milking machine. With this objective, 100 Murciano-Granadina goats with similar parturition dates (4 ± 1 postpartum weeks) were divided into 2 similar groups (50 goats per group) according their parity, milk yield, mammary gland health status, and milking duration (after a 15-d pre-experimental period). Throughout the experimental period (7 mo), one group was milked using SP and the other group using AP. Every month, samplings were carried out of the variables related to sanitary status of the mammary gland, milk fractioning, milking duration, average and maximum milk flows, milk composition, teat end status, vacuum fluctuations at the short milk tube during milking, and pulsation tests. Results showed that the use of SP in a high pipeline milking machine showed higher vacuum fluctuations without involving any difference in the other variables studied. We concluded that the use of AP in the milking of Murciano-Granadina goats offers little improvement of the milking performance and increases the investment required for installation and maintenance of the milking machines.

Goal-directed therapy in cytoreductive surgery with hyperthermic intraperitoneal chemotherapy: a prospective observational study.

BACKGROUND: Cytoreductive surgery (CRS) with Hyperthermic Intraperitoneal Chemotherapy (HIPEC) in peritoneal carcinomatosis treatment causes significant hemodynamic, metabolic, and hematological alterations. Studies on the anesthetic intraoperative management are heterogeneous and scarce. There is a great heterogeneity in the anesthetic management of CRS and HIPEC. The aim of this study is to analyze perioperative hemodynamic goal-directed management and to evaluate the complications arisen until the seventh postoperative day. METHODS: Prospective, observational study of all CRS and HIPEC patients from March 2014 to May 2017. Hemodynamic and clinical parameters were registered during surgery and the first 3 postoperative days. We correlated intraoperative data with the postoperative course until the seventh day. RESULTS: A total of 92 patients were included in the study (age 58.5 ± 10.9 years, 47% colorectal carcinoma, and 38% ovarian carcinoma). Peritoneal Carcinomatosis Index (PCI) (median and ranges) was 10 [0-39]. Cardiac Index (CI) 3.15 l/min-1/m-2 [1.79-5.60]) and Systolic Volume Variation (SVV) (10% [3%-17%]) remained within the values of normality in all surgery phases. A large difference was observed between the minimum and maximum ranges of fluid therapy administered (median 9.8 ml/kg/h [5.3-24.3]), showing a great interindividual variation in the fluids requirement. A direct relationship was observed between PCI and surgery duration, fluid therapy, and intraoperative transfusion percentage (p < 0.02). CONCLUSIONS: There is a great variability in the intraoperative fluid therapy needs of the patients. SVV monitoring makes it possible to adjust the fluid therapy needs in each surgery phase. The use of a hemodynamic goal-directed anesthetic protocol in CRS and HIPEC enables to individually adjust the fluid therapy, avoiding over-hydration and ensuring hemodynamic stability in all surgery phases.