Nanostructured lipid carriers loaded with essential oils: a strategy against SARS-CoV-2.

This work aimed to investigate the effectiveness of Lippia sidoides and Syzygium aromaticum essential oils (EOs) encapsulated in nanostructured lipid carriers (NLCs) as SARS-CoV-2 inhibitors through virucidal activity assessment. We developed anionic and cationic NLCs loaded with the EOs and assessed their physicochemical properties and SARS-CoV-2 virucidal activity, focusing on the effects of EO type and the NLCs composition. The NLCs exhibited particle sizes of 141.30 to 160.53 nm for anionic and 109.30 to 138.60 nm for cationic types, with PDIs between 0.16 and 0.25. High zeta potentials (>29.0 in modulus) indicated stable formulations. The NLCs effectively encapsulated the EOs, achieving encapsulation efficiencies between 84.6 to 100% w/w of marker compound. The EOs-loaded NLCs reduced the SARS-CoV-2 virion count, exceeding 2 logs over the control. NLCs loaded with Lippia sidoides and Syzygium aromaticum EOs represent an innovative strategy for combating SARS-CoV-2.

Modeling the relationship between heat stress, feed intake, and day relative to calving in nonlactating dairy cows.

Heat stress (HS) during the dry period can affect animal welfare, health, dry matter intake (DMI), and milk production in the subsequent lactation, which will negatively affect the profitability of dairy farms. In this study, the objective was to model the changes in DMI in pregnant nonlactating heat-stressed dairy cows with or without access to evaporative cooling systems. A database was built, composed of individual DMI records from 244 pregnant nonlactating dairy cows from an average -29.3 d (range: -42 to -21 d; SD: ±7.54 d) to -1 d relative to calving (DRC) and housed in environmental conditions in which temperature-humidity index (THI) ranged from 58.4 to 83.3, with or without access to evaporative cooling systems. Generalized additive mixed-effects models were used to describe the relationships of DMI with HS and DRC. Changes in DMI with the increase in THI and the progression of pregnancy in cows with or without evaporative cooling systems were estimated using differential equations. On average, cows housed in barns without evaporative cooling systems had a reduction in DMI of 1.30 kg/d and increased rectal temperature in 0.22°C in relation to those housed in barns with evaporative cooling systems. Dry matter intake decreased as THI increased, but the reduction was greater for noncooled cows as THI values increased. In addition, regardless of the THI, DMI started to decrease at -14 DRC for cooled cows, whereas for noncooled cows it already started at -30 DRC, relative to the previous days evaluated. The intensity of the reduction was lesser for cows that had access to evaporative cooling systems or were in the dry period in May to June as compared with those that were in the dry period in July to August or September to October. The models generated in this study, which include environmental variables, should lead to more accurate predictions of DMI during HS that can be used to formulate diets to meet the needs of the late pregnant cow because it is possible to predict changes in DMI as the heat load and DRC change. Such models are also expected to help dairy nutritionists to decide when and how to apply the dietary strategies available to attenuate the reductions in DMI with the intensity of HS and progression of pregnancy.

Assessing milk response to different combinations of branched-chain volatile fatty acids and valerate in Jersey cows.

Some cellulolytic bacteria require 1 or more branched-chain volatile fatty acids (BCVFA) for the synthesis of branched-chain AA and branched-chain long-chain fatty acids because they are not able to uptake branched-chain AA or lack 1 or more enzymes to synthesize branched-chain AA de novo. Supplemental BCVFA and valerate were included previously as a feed additive that was later removed from the market; these older studies and more current studies have noted improvements in neutral detergent fiber digestibility and milk efficiency. However, most studies provided a single BCVFA or else isobutyrate (IB), 2-methylbutyrate (MB), isovalerate, and valerate altogether without exploring optimal combinations. Our objective was to determine a combination of isoacids that is optimal for milk production. Sixty (28 primiparous and 32 multiparous) lactating Jersey cows (106 ± 54 days in milk) were blocked and assigned randomly to either a control (CON) treatment without any isoacids, MB [12.3 mmol/kg dry matter (DM)], MB + IB (7.7 and 12.6 mmol/kg DM of MB and IB, respectively), or all 4 isoacids (6.2, 7.3, 4.2, and 5.1 mmol/kg DM of MB, IB, isovalerate, and valerate, respectively). Cattle were fed the CON treatment for a 2-wk period, then were assigned randomly within a block to treatments for 8 wk (n = 15). There was a trend for an interaction of supplement and parity for milk components. There were no differences in components for primiparous cows, whereas MB + IB tended to increase protein concentration by 0.04 and 0.08 percentage units in multiparous cows compared with the CON and MB treatments, respectively. Feeding MB + IB increased fat concentration by 0.23 to 0.31 percentage units compared with all other treatments in multiparous cows. Milk yield and dry matter intake (DMI) did not change with treatment. Treatment interacted with week for milk net energy for lactation/DMI; MB + IB tended to increase milk net energy of lactation/DMI by 0.10 Mcal/kg compared with MB and approached a trend for CON, mainly during the early weeks of the treatment period, whereas differences decreased during the last 2 wk of the treatment period. Cows fed MB had the highest 15:0 anteiso fatty acids in the total milk fatty acid profile, which was greater than that for CON or MB + IB cows, but not cows supplemented with isoacids. Cows fed MB alone had the numerically lowest milk net energy for lactation/DMI. The combination of MB + IB appeared optimal for increasing feed efficiency in our study and was not at the expense of average daily gain. Further research is needed for evaluating how potential changes in supplemental isoacid dosage should vary under differing dietary conditions.

Modeling the effects of heat stress in animal performance and enteric methane emissions in lactating dairy cows.

Heat stress (HS) negatively affects dry matter intake (DMI), milk yield (MY), feed efficiency (FE), and free water intake (FWI) in dairy cows, with detrimental consequences to animal welfare, health, and profitability of dairy farms. Absolute enteric methane (CH4) emission, yield (CH4/DMI), and intensity (CH4/MY) may also be affected. Therefore, the goal of this study was to model the changes in dairy cow productivity, water intake, and absolute CH4 emissions, yield, and intensity with the progression (days of exposure) of a cyclical HS period in lactating dairy cows. Heat stress was induced by increasing the average temperature by 15°C (from 19°C in the thermoneutral period to 34°C) while keeping relative humidity constant at 20% (temperature-humidity index peaks of approximately 83) in climate-controlled chambers for up to 20 d. A database composed of individual records (n = 1,675) of DMI and MY from 82 heat-stressed lactating dairy cows housed in environmental chambers from 6 studies was used. Free water intake was also estimated based on DMI, dry matter, crude protein, sodium, and potassium content of the diets, and ambient temperature. Absolute CH4 emissions was estimated based on DMI, fatty acids, and dietary digestible neutral detergent fiber content of the diets. Generalized additive mixed-effects models were used to describe the relationships of DMI, MY, FE, and absolute CH4 emissions, yield, and intensity with HS. Dry matter intake and absolute CH4 emissions and yield reduced with the progression of HS up to 9 d, when it started to increase again up to 20 d. Milk yield and FE reduced with the progression of HS up to 20 d. Free water intake (kg/d) decreased during the exposure to HS mainly because of a reduction in DMI; however, when expressed in kg/kg of DMI it increased modestly. Methane intensity also reduced initially up to d 5 during HS exposure but then started to increase again following the DMI and MY pattern up to d 20. However, the reductions in CH4 emissions (absolute, yield, and intensity) occurred at the expense of decreases in DMI, MY, and FE, which are not desirable. This study provides quantitative predictions of the changes in animal performance (DMI, MY, FE, FWI) and CH4 emissions (absolute, yield, and intensity) with the progression of HS in lactating dairy cows. The models developed in this study could be used as a tool to help dairy nutritionists to decide when and how to adopt strategies to mitigate the negative effects of HS on animal health and performance and related environmental costs. Thus, more precise and accurate on-farm management decisions could be taken with the use of these models. However, application of the developed models outside of the ranges of temperature-humidity index and period of HS exposure included in this study is not recommended. Also, validation of predictive capacity of the models to predict CH4 emissions and FWI using data from in vivo studies where these variables are measured in heat-stressed lactating dairy cows is required before these models can be used.

Characteristics of mammary secretions collected from infected and uninfected primigravid dairy heifer mammary glands.

Intramammary infections (IMI) in primigravid dairy heifers can affect mammary growth and development, which can reduce first-lactation milk yield. Detection of IMI in heifers most often involves the use of culture-based methods that are not often used in production dairy settings given their labor- and time-consuming nature. The objective of this study was to determine whether mammary secretion somatic cell count (SCC) and viscosity were associated with the infection status of primigravid heifer mammary glands. A total of 270 heifers from a single farm were used, selected based on the farmer's willingness to participate. The study was conducted from June to October 2020. Mammary secretion samples were aseptically collected from a randomly selected quarter of each heifer at 75 d prepartum (75PP), and another quarter of each heifer was sampled at 35 d prepartum (35PP). The remaining 2 quarters of each heifer were not examined. Mammary secretion samples underwent bacteriological examination to determine IMI status and quantitative SCC measurement and were also assessed for secretion viscosity based on visual observation. Prevalence of IMI was 26% (69/270) and 28% (71/255) at 75 and 35 d prepartum, respectively. Uninfected secretion samples had 133.2 [95% confidence interval (CI): 16.8 to >999.9] times greater odds to be thick compared with samples infected with a major pathogen, and 14.4 (95% CI: 8.5 to 24.1) times greater odds to be thick compared with samples infected with non-aureus staphylococci (NAS). The mean secretion SCC of uninfected quarters (6.04 ± 0.03 log10 cells/mL) was significantly lower than that of secretions collected from quarters infected with Staphylococcus chromogenes (6.34 ± 0.04 log10 cells/mL), other NAS species (6.28 ± 0.10 log10 cells/mL), or a major pathogen (6.73 ± 0.08 log10 cells/mL). These results indicate that mammary secretion viscosity and SCC measurement may be useful tools in identifying primigravid heifer quarters with IMI. The ability to evaluate viscosity at time of sampling may be a useful strategy that could be incorporated into interventions designed to diminish the negative effects of prepartum IMI on lactational performance.

Effects of prepartum stocking density and a blind on physiological biomarkers, health, and hygiene of transition Holstein dairy cows.

Many dairy cows succumb to disease after calving. Disease risk may be affected by the cows' social environment and ability to perform maternal behaviors. In nature, cattle isolate from others and find seclusion to give birth; these behaviors may be limited in indoor group pens and could potentially affect the cows' ability to cope. The aim was to determine whether stocking density and a physical blind in prepartum bedded-pack group pens affected physiological biomarkers, disease risk, and hygiene of peripartum dairy cows. A randomized complete block designed with a 2 × 2 factorial arrangement of treatments was used. Approximately 3 wk before calving, 374 cows (primiparous = 127; multiparous = 247) were assigned randomly to 1 of 4 treatment pens with the following 2 factors: (1) high versus low stocking density (7.7 to 12.9 m2 vs. 15.4 to 25.8 m2 lying space per cow), and (2) presence or absence of a blind. The blind was created using plastic road barriers and plywood, a steel gate, and shade cloth. After calving, cows were moved into a freestall pen and milked 3 times per day. Blood was sampled on the day of enrollment (baseline; -24 d) and approximately -14, -7, 3, 7, 10, and 14 d relative to calving, to measure inflammatory (haptoglobin) and metabolic (nonesterified fatty acids and ß-hydroxybutyrate) biomarkers. ß-Hydroxybutyrate (≥1.2 mmol/L) was used to diagnose subclinical ketosis. Vaginal discharge was scored 3, 7, 10, and 14 d after calving, to diagnose metritis (none, mild, or severe). Hygiene was scored on -24, -14, and -7 d before calving. Before calving, haptoglobin was lower in pens with a blind. After calving, cows in pens with low stocking density before calving tended to be at greater odds of being diagnosed with metritis. Cows were more likely to have poorer hygiene scores in high stocking density pens. No treatment effects were detected for pre- and postpartum nonesterified fatty acids, ß-hydroxybutyrate, postpartum haptoglobin, or subclinical ketosis. Results suggest that the provision of a blind and lower stocking density may be beneficial for reducing inflammation before calving. However, low prepartum stocking density might increase the odds of metritis after calving. Although the reason for this paradox is unclear, the effects of prepartum stocking density may require further exploration.

The effect of stocking density and a blind on the behavior of Holstein dairy cows in group maternity pens. Part II: Labor length, lying behavior, and social behavior.

In natural settings, dairy cows separate from the herd to give birth. When kept indoors, seeking isolation before calving may be restricted and may depend on space and resources provided in maternity housing. The effect of group maternity pens on behavior around calving and labor progress is unknown. Therefore, the objective of this study was to determine the effects of stocking density and provision of a blind in group bedded pack maternity pens on lying and social behavior as well as length of labor of preparturient dairy animals. The study was conducted as a complete randomized block design with a 2 × 2 factorial arrangement of treatments including stocking density and presence or absence of a blind, resulting in a total of 4 treatments: (1) high stocking density (7.7-12.9 m2 lying space/cow) with a blind, (2) low stocking density (15.4-25.8 m2) with a blind, (3) high stocking density without a blind, and (4) low stocking density without a blind. A total of 127 primiparous heifers and 247 multiparous cows were housed in mixed-parity groups from approximately 3 wk before and immediately after calving. During the 4 h before calving, lying behavior (lying time and bouts) was collected automatically using accelerometers, and social behavior (agonistic interactions, allogrooming, and attention from other cows), stage II labor duration, and frequency of position change during stage II labor were collected using video. Lying behavior was collected with accelerometers. Regardless of treatment, lying time and bouts increased as calving approached. Cows and heifers performed more lying bouts in low stocking density pens compared with high stocking density pens. Agonistic interactions and allogrooming were not different between treatments. Other cows spent more time paying attention to focal animals regardless of stocking density as calving approached, but time spent paying attention was reduced by the presence of a blind during h -2 before calving. The hazard of calving unassisted was greater for cows and heifers in low stocking density pens with a blind compared with all other treatments. Further, animals in pens with a blind tended to change positions fewer times during stage II labor. These results suggest that providing a blind in group maternity pens may improve the calving environment for cows and heifers and, in combination with low stocking density, may reduce the amount of time spent in labor.

Organization of mammary blood vessels as affected by mammary parenchymal region and estradiol administration in Holstein heifer calves.

Mammary blood flow is central to mammary growth, development, and productivity, but the development of the vasculature network is poorly understood. The objective of this study was to determine how the vascular system adapts to mammary growth by inducing different levels of mammary growth and examining 2 regions of mammary parenchymal tissue. Holstein heifer calves (n = 12) received daily injections on the days immediately preceding euthanasia at 82 d of age. Treatments were control (CON), short-term estradiol (STE), and long-term estradiol (LTE). The CON calves received corn oil injections, the STE calves received 9 injections of corn oil followed by 3 injections of estradiol, and the LTE calves received 12 estradiol injections. Mammary tissues were collected from the center and edge parenchymal regions of all right rear mammary glands to quantify the tissue area of various tissue structures, the percentage of proliferating epithelial cells, and the number and form of blood vessels. Results showed that LTE calves had a greater tissue area occupied by epithelium than CON and STE calves, and the epithelial area in CON and STE calves was similar. Edge parenchyma had a greater percentage of proliferating epithelial cells than center parenchyma across all treatment groups. In the edge region, LTE calves had the greatest percentage of proliferating epithelial cells, coinciding with greater epithelial area. The number of blood vessels per unit of tissue area was greater in center than in edge parenchyma; the corresponding vessel surface area per unit of tissue area followed the same pattern. Mammary blood vessel measures were not markedly influenced by estradiol treatment. These results highlight the marked difference in the number and organization of blood vessels in different mammary parenchyma regions but indicate that the effects of estradiol on stimulating mammary epithelial proliferation does not directly translate to increasing numbers of blood vessels.

Administration of internal teat sealant in primigravid dairy heifers at different times of gestation to prevent intramammary infections at calving.

Intramammary infections (IMI) are common in primigravid dairy heifers and can negatively affect future milk production. Bismuth subnitrate-based internal teat sealants (ITS) have been used to prevent prepartum IMI in dairy heifers by creating a physical barrier within the teat, preventing pathogens from entering the gland, though determination of when to administer ITS in heifers has yet to be investigated. The objectives of this study were to determine if administration of ITS in primigravid heifers reduced the odds of IMI at calving and if administration of ITS at different stages of gestation (75 vs. 35 d prepartum) affected the odds of IMI at calving. A total of 270 heifers were used at a single farm. One quarter of each heifer was randomly chosen to be aseptically sampled and administered ITS 75 d prepartum (ITS75), another quarter of each heifer was sampled and received ITS 35 d prepartum (ITS35), whereas the remaining 2 quarters of each heifer served as control quarters (CON) and were not sampled before calving. Within 12 h of calving, aseptic colostrum samples were collected from all quarters to determine quarter infection status. When an IMI was caused by mastitis pathogens other than non-aureus staphylococci (NAS), CON quarters were 3 times [95% confidence interval (CI): 1.4-6.3] and 2.5 times (95% CI: 1.2-4.9) more likely to be infected at calving than ITS75 and ITS35 quarters, respectively. For IMI with NAS, CON quarters were 5.8 (95% CI: 3.2-10.5) and 6.4 (95% CI: 3.4-12.0) times more likely to be infected than ITS75 and ITS35 quarters, respectively. Odds of IMI at calving was similar between ITS75 and ITS35 quarters for both NAS (odds ratio = 0.9) and other pathogens (odds ratio = 1.2). Results indicate that ITS administration at either 75 and 35 d prepartum reduced IMI prevalence at calving in primigravid dairy heifers. Farm specific factors may influence prevalence and timing of heifer IMI and earlier administration of ITS provides an extended period of protection for the developing gland.

Isoflavones improve collagen I and glycosaminoglycans and prevent bone loss in type 1 diabetic rats.

Objective: The objective of this study was to evaluate the action of soy isoflavones (ISO) and 17ß-estradiol on collagen I (CollI) and sulfated glycosaminoglycans (GAGs) in the bone matrix of diabetic rats.Methods: Sixty adult female rats (Rattus norvegicus albinus) underwent ovariectomy, and then were randomized into six groups of 10 animals each: GI, sham control ovariectomized animals; GII, sham control diabetic (DM) ovariectomized animals; GIII, control ovariectomized animals receiving propylene glycol vehicle; GIV, control ovariectomized DM animals receiving propylene glycol vehicle; GV, ovariectomized DM animals treated with ISO (150 mg/kg by gavage); and GVI, ovariectomized DM animals treated with estrogen (17ß-estradiol, 10 mg/kg, subcutaneously). 17ß-Estradiol was used as a positive control when compared with ISO. To obtain significant depletion of the estrogen levels and subsequent bone loss, a postsurgical period of 90 days was observed. Treatments occurred during 30 consecutive days. After euthanasia, shafts of the animals' femurs were immersed in liquid nitrogen for molecular biology analysis, and the distal femurs were removed and processed for paraffin embedding.Results: ISO (GV) and 17ß-estradiol (GVI) improved bone formation, increasing GAGs and CollI formation when compared to the control group (GIV) (p < 0.05).Conclusions: ISO and 17ß-estradiol contribute to the decrease of bone loss in diabetic rats.

Meta-analysis of the relationship between milk trans-10 C18:1, milk fatty acids <16 C, and milk fat production.

The economic value of milk fat and its responsiveness to management strategies provides strong interest in maximizing milk fat production by minimizing occurrence of biohydrogenation-induced milk fat depression (BH-MFD) and maximizing de novo synthesized fatty acids (FA). Tools that allow a timely diagnosis of BH-MFD would improve nutritional management. Specific milk FA or FA categories correlate to milk fat concentration and are of interest for diagnosing the cause of changes in milk fat concentration. The objective of the current study was to characterize the relationship between milk fat concentration and trans-10 C18:1, a proxy for BH-MFD, and FA <16 carbons that originate solely from de novo lipogenesis using a meta-analysis approach that used data from the literature and unpublished Penn State experiments. Prior to the meta-analysis, the effect of FA methylation method on milk FA profile was tested to determine potential bias between papers. There was no difference between sodium methoxide, acid, and acid-base methylation methods on trans-10 C18:1 concentration, but acid methods resulted in loss of short-chain FA. The relationship between trans-10 C18:1 and milk fat percentage was investigated using a 2-component model, where one component described the fraction unresponsive to BH-MFD and the other described a responsive fraction that is exponentially related to trans-10 C18:1. The 2 fractions where characterized utilizing a Bayesian hierarchical model accounting for between-study variability. The model was defined by the function f(x, θ1, θ2, θ3) = θ1 + θ2exp(θ3), where the unresponsive θ1 fraction was 2.15 ± 0.09%, the responsive θ2 fraction was 1.55 ± 0.08%, and the exponential term θ3 was -0.503 ± 0.07 (posterior mean ± posterior standard deviation from the Bayesian hierarchical model). A Lin's concordance correlation coefficient of 0.67 suggested good agreement between observations and predictions from the Bayesian hierarchical model, computed only with the model's mean population parameters. There was a linear relationship between milk fat concentration and FA <16 C as a percentage of total FA (intercept = 2.68 ± 0.237 and slope = 0.043 ± 0.011; coefficient of determination = 0.31). The relationship between milk FA <16 C and milk fat concentration is weaker than what has been published, likely because multiple factors can reduce de novo FA without reducing milk fat and the broad range of diets present in the literature.

Short communication: Heat stress does not affect induced luteolysis in Holstein cows.

Heat stress (HS) has deleterious effects on bovine reproduction, including prolongation of the luteal phase in Holstein cows, perhaps due to compromised luteolysis. The objective was to characterize effects of HS on luteolytic responses of nonlactating Holstein cows given 25 or 12.5 mg of PGF2α on d 7 of the estrous cycle. Cows were randomly distributed into 2 environments: thermoneutral (n = 12; 25°C) or HS (n = 12; 36°C). In each environment, cows were treated with 2 mL of saline, 25 or 12.5 mg of PGF2α (n = 4 cows per group). The HS environment induced a significant increase in rectal temperature and respiratory rate compared with the thermoneutral environment. Heat stress did not have significant effects on luteolytic responses or circulating progesterone concentrations. Rapid and complete luteolysis occurred in all cows given 25 mg of PGF2α and in 4 of 8 cows given 12.5 mg; the other 4 cows given 12.5 mg had partial luteolysis, with circulating progesterone concentrations initially suppressed, but subsequently rebounding. Therefore, we conclude that HS does not change corpus luteum sensitivity to PGF2α.

Microbial ecology and predicted metabolic pathways in various oral environments from patients with acute endodontic infections.

AIM: To assess in a cross-sectional clinical study the effect of antibiotics on the diversity, structure and metabolic pathways of bacterial communities in various oral environments in patients with acute primary infections. METHODOLOGY: Samples of saliva (SA), supragingival biofilm (SB) and from the pulp cavity (PC) were collected from teeth with acute primary infections and then grouped according to previous use of antibiotics (NoAtb = no antibiotics [n = 6]; Atb = antibiotics [n = 6]). DNA sequencing was conducted using MiSeq (Illumina, San Diego, CA, USA). The V1-V3 hyper-variable region of the 16S rRNA gene was amplified. A custom Mothur pipeline was used for 16S rRNA processing. Subsequent analyses of the sequence dataset were performed in R (using vegan, phyloseq and ggplot2 packages) or QIIME. RESULTS: Twelve patients aged from 22 to 56 years were recruited. Participants in the Atb group had taken the beta-lactamics amoxicillin (5/6) or cephalexin (1/6) for 2-3 days. A total of 332 bacterial taxa (OTUs) were identified, belonging to 120 genera, 60 families and nine phyla. Firmicutes (41%) and Bacteroidetes (38%) were the most abundant phyla in all samples. Taxa clustered significantly by oral site (PCoA analysis; P < 0.05, ANOSIM). Use of antibiotics had little effect on this clustering. However, SA, SB and PC had different degrees of richness, diversity and evenness. The greatest diversity was observed in SB samples and the least diversity was observed in PC samples. Metabolic prediction identified 163 pathways and previous use of antibiotics had a major effect on the estimated functional clustering in SA and PC samples. CONCLUSION: The ecological niche had a strong influence on the bacterial content of samples from various oral sites. Previous exposure to antibiotics may exert an effect on the phylogenetic composition of SA. Metabolic pathways appear to be modulated by antimicrobial agents in SA and PC samples. The dynamics of host/microbial interactions in the apical region and the functional ecology of the infected pulp cavity should be revisited.

Bioprocess strategies for enhancing the outdoor production of Nannochloropsis gaditana: an evaluation of the effects of pH on culture performance in tubular photobioreactors.

A priority of the industrial applications of microalgae is the reduction of production costs while maximizing algae biomass productivity. The purpose of this study was to carry out a comprehensive evaluation of the effects of pH control on the production of Nannochloropsis gaditana in tubular photobioreactors under external conditions while considering the environmental, biological, and operational parameters of the process. Experiments were carried out in 3.0 m3 tubular photobioreactors under outdoor conditions. The pH values evaluated were 6.0, 7.0, 8.0, 9.0, and 10.0, which were controlled by injecting pure CO2 on-demand. The results have shown that the ideal pH for microalgal growth was 8.0, with higher values of biomass productivity (Pb) (0.16 g L-1 d-1), and CO2 use efficiency ([Formula: see text]) (74.6% w w-1); [Formula: see text]/biomass value obtained at this pH (2.42 [Formula: see text] gbiomass-1) was close to the theoretical value, indicating an adequate CO2 supply. At this pH, the system was more stable and required a lower number of CO2 injections than the other treatments. At pH 6.0, there was a decrease in the Pb and [Formula: see text]; cultures at pH 10.0 exhibited a lower Pb and photosynthetic efficiency as well. These results imply that controlling the pH at an optimum value allows higher CO2 conversions in biomass to be achieved and contributes to the reduction in costs of the microalgae production process.

Is the inferior occlusal plane a reliable anatomic landmark for inferior alveolar nerve block? A study on dry mandibles of Brazilian adults.

PURPOSE: To investigate the relationship between the mandibular foramen (MF) and the inferior occlusal plane (IOP) of anatomic specimens, in order to guide clinicians in the inferior alveolar nerve block (IANB). METHOD: Eighty-two dry mandibles (totally or partially toothed) of Brazilian adults were selected, totalizing 66 left and 66 right antimeres. A flat plate of rigid PVC plastic was placed on the occlusal surfaces of posterior teeth and pressure was then manually applied to simulate the IOP. Following, the perpendicular distance from the IOP to the lowest point of the MF was measured by the use of a digital caliper, adopting negative values inferiorly and positive values superiorly. RESULTS: Regarding the antimeres, the majority of the lowest limits of the MF was below the IOP (right: 86.4%, left: 81.8%) with a mean of -4.2mm (±4.8) in the right and -4.1mm (±4.8) in the left ones, without any statistical significance. Considering both sexes, no statistically significant difference was obtained between them. CONCLUSION: The IOP seems not to be a good anatomical landmark to localize the MF and, its turn, to guide clinicians during the IANB for Brazilian adults, given the marked topographic variations.

Quantifying the effects of mastitis on the reproductive performance of dairy cows: A meta-analysis.

The objective of this study was to conduct a meta-analysis to quantitatively characterize the effects of mastitis on the reproductive performance of dairy cows as well as to identify factors that interact with this relationship. A total of 29 publications were identified that contributed 24, 41, 27, 38, and 13 trial results to the meta-analysis of how mastitis is related to time to first service (TFS), days open (DO), services per conception (SPC), pregnancies per insemination at first service (FSP/AI) and pregnancy loss (PL), respectively. The meta-analyses were conducted using multilevel linear mixed-effects models. Overall, high levels of heterogeneity were present and meta-regression models only explained a small amount of heterogeneity. Results suggest that cows with mastitis pre-first insemination experience, on average, an additional 13.29 d to first service [95% confidence interval (CI): 6.64, 19.95] when compared with cows with no mastitis in the same time period. Moreover, in relation to cows with no mastitis in the same time period, 22.34 additional DO (95% CI: 12.89, 31.79) were estimated, on average, for cows with clinical mastitis at pre-insemination leading to conception. Additionally, 32.41 added DO (95% CI: 20.58, 44.25) were estimated, on average, for cows with clinical mastitis at pre- or post-insemination leading to conception compared with cows with no mastitis in the same time period. Finally, 20.03 additional DO (95% CI: 3.11, 36.95) were estimated, on average, for cows with subclinical mastitis pre- or post-insemination leading to conception compared with cows with no mastitis in the same time period. Effect size estimates from the meta-regression models for SPC, in relation to cows with no mastitis in the same time period, suggest that, on average, SPC increases by 0.46 inseminations (95% CI: 0.30, 0.62) for a cow experiencing mastitis pre-insemination leading to conception. When mastitis occurs either pre- or post-insemination leading to conception, SPC is expected to increase, on average, by 0.72 inseminations (95% CI: 0.48, 0.95) compared with cows with no mastitis in the same time period. The estimated effect sizes for FSP/AI suggest a risk ratio of conceiving at first insemination of 0.90 (95% CI: 0.82, 0.99) for cows with mastitis diagnosed pre-first insemination with respect to cows with no mastitis in the same time period, and a risk ratio of 0.79 (95% CI: 0.72, 0.86) for cows diagnosed with mastitis either pre- or post-first insemination with respect to cows with no mastitis in the same time period. Publication bias was identified in 4 of the meta-analysis models (TFS, DO, SPC, and FSCR), but no influential trials were identified in any models; the reliability of the meta-analysis results should be interpreted carefully keeping these limitations in mind. Further meta-regression analysis would be valuable as additional studies are published that report other potential sources of heterogeneity.

Predicting milk protein responses and the requirement of metabolizable protein by lactating dairy cows.

The objective of this study was to develop a modeling framework to predict milk protein yield responses to varying metabolizable protein (MP) supplies and to determine the requirement of MP by lactating dairy cows. The logistic curve was used to model milk protein yield while accounting for a variable efficiency of MP utilization and between-study variability. Models were developed with databases from 2 recently published meta-analyses and based on either total MP supply or MP supply available for milk production. All models provided reasonable fit to data, with root mean square prediction error ranging from 18 to 20% of the average milk protein yield. The estimated horizontal asymptotes were 1.17 (posterior SD = 0.02) and 1.55 (posterior SD = 0.06) in the 2 databases, suggesting that the limiting milk protein yield, as MP supply increases, converges to 1.17 or 1.55 kg/d in the environments determined by the 2 databases. The observed efficiencies ranged from 0.75 to 0.18 when total MP supply was used as the denominator and above 1 to 0.24 when the MP supply available for milk production was used as the denominator. The predicted efficiencies were in good agreement with the data, decreasing nonlinearly with the MP supply. The MP requirement was calculated with a function constructed with the inverse of the logistic model and modified at regions of maximum marginal efficiency and minimum second derivative. This strategy assumes that the MP solution, or the MP needed to predict a given protein yield in the fitted logistic curve, determines the MP requirement for maintenance and lactation. Requirements calculated with the independent variable as total MP supply refer to the total requirement of maintenance plus lactation, whereas the requirement from models based on MP supply available for milk production are referent to the MP required only for lactation. The requirements were, on average, slightly smaller than the ones predicted by the current Northern American feeding system for dairy cows at lower protein yields and greater than currently recommended at high yields.

Predicting manure volatile solid output of lactating dairy cows.

Organic matter (OM) in livestock manure consisting of biodegradable and nonbiodegradable fractions is known as volatile solids (VS). According to the Intergovernmental Panel on Climate Change (IPCC) Tier 2 guidelines, methane produced by stored manure is determined based on VS. However, only biodegradable OM generates methane production. Therefore, estimates of biodegradable VS (dVS; dVS = VS - lignin) would yield better estimates of methane emissions from manure. The objective of the study was to develop mathematical models for estimating VS and dVS outputs of lactating dairy cows. Dry matter intake, dietary nutrient contents, milk yield and composition, body weight, and days in milk were used as potential predictor variables. Multicollinearity, model simplicity, and random study effects were taken into account during model development that used 857 VS and dVS measurements made on individual cows (kg/cow per day) from 43 metabolic trials conducted at the USDA Energy and Metabolism laboratory in Beltsville, Maryland. The new models and the IPCC Tier 2 model were evaluated with an independent data set including 209 VS and dVS measurements (kg/cow per day) from 2 metabolic trials conducted at the University of California, Davis. Organic matter intake (kg/d) and dietary crude protein and neutral detergent fiber contents (% of dry matter) were significantly associated with VS. A new model including these variables fitted best to data. When evaluated with independent data, the new model had a root mean squared prediction error as a percentage of average observed value (RMSPE) of 12.5%. Mean and slope biases were negligible at <1% of total prediction bias. When energy digestibility of the diet was assumed to be 67%, the IPCC Tier 2 model had a RMSPE of 13.7% and a notable mean bias for VS to be overpredicted by 0.4 kg/cow per day. A separate model including OM intake as well as dietary crude protein and neutral detergent fiber contents as predictor variables fitted best to dVS data and performed well on independent data (RMSPE = 12.7%). The Cornell Net Carbohydrate and Protein System model relying on fat-corrected milk yield and body weight more successfully predicted dry matter intake (DMI; RMSPE = 14.1%) than the simplified (RMSPE = 16.9%) and comprehensive (RMSPE = 23.4%) models to predict DMI in IPCC Tier 2 methodology. New models and the IPCC Tier 2 model using DMI from the Cornell Net Carbohydrate and Protein System model predicted VS (RMSPE = 17.7-19.4%) and dVS (RMSPE = 20%) well with small systematic bias (<10% of total bias). The present study offers empirical models that can accurately predict VS and dVS of dairy cows using routinely available data in dairy farms and thereby assist in efficiently determining methane emissions from stored manure.

Antimicrobial activity of crude extracts from actinomycetes against mastitis pathogens.

The emergence of antimicrobial resistance to commonly used antibiotics has necessitated the development of new antimicrobial products. Crude extracts produced by actinomycetes contain antimicrobial metabolites that can inhibit bacterial growth. The objective of our study was to evaluate the antimicrobial activity of crude extracts (Caat1-54 and CaatP5-8) produced by actinomycetes against isolates of Staphylococcus aureus, Staphylococcus chromogenes, Streptococcus dysgalactiae, and Streptococcus uberis, which were obtained from the milk of cows affected by mastitis in 23 dairy herds. Twenty isolates of each bacterial species were used to define minimum inhibitory concentrations (MIC) of both crude extracts and ceftiofur (positive control). The MIC50 and MIC90 were defined at the concentration required to inhibit the growth of 50 and 90% of bacterial isolates tested, respectively. The MIC results were evaluated by survival analysis. Staphylococcus aureus isolates presented MIC90 of Caat 1-54 ≥6.25 µg/mL, ceftiofur ≥12.5 µg/mL, and Caat P5-8 ≥100 µg/mL. Streptococcus uberis presented MIC90 of ceftiofur ≥0.39 µg/mL, Caat 1-54 ≥50 µg/mL, and Caat P5-8 ≥100 µg/mL. Staphylococcus chromogenes isolated from subclinical mastitis presented MIC90 of Caat 1-54 ≥0.78 µg/mL and ceftiofur and Caat P5-8 of ≥6.25 and ≥100 µg/mL, respectively. Streptococcus dysgalactiae isolated from clinical mastitis presented similar MIC90 values between antimicrobials tested (ceftiofur, Caat 1-54, and Caat P-58), but these values (≥100 µg/mL) were higher than the values obtained from other pathogens evaluated in the present study. Our results indicate that Caat 1-54 and Caat P5-8 crude extracts present in vitro antimicrobial activity against isolates of Staph. aureus, Staph. chromogenes, Strep. dysgalactiae, and Strep. uberis isolated from clinical and subclinical mastitis.

Short communication: Urea hydrolysis in dairy cattle manure under different temperature, urea, and pH conditions.

The objective of the study was to quantify the rate of urea hydrolysis in dairy cattle manure under different initial urea concentration, temperature, and pH conditions. In particular, by varying all 3 factors simultaneously, the interactions between them could also be determined. Fresh feces and artificial urine solutions were combined into a slurry to characterize the rate of urea hydrolysis under 2 temperatures (15°C and 35°C), 3 urea concentrations in urine solutions (500, 1,000, and 1,500 mg of urea-N/dL), and 3 pH levels (6, 7, and 8). Urea N concentration in slurry was analyzed at 0.0167, 1, 2, 4, 6, 8, 12, 16, 20, and 24 h after initial mixing. A nonlinear mixed effects model was used to determine the effects of urea concentration, pH, and temperature treatments on the exponential rate of urea hydrolysis and to predict the hydrolysis rate for each treatment combination. We detected a significant interaction between pH and initial urea level. Increasing urea concentration from 1,000 to 1,500 mg of urea-N/dL decreased the rate of urea hydrolysis across all pH levels. Across all pH and initial urea levels, the rate of urea hydrolysis increased with temperature, but the effect of pH was only observed for pH 6 versus pH 8 at the intermediate initial urea concentration. The fast rates of urea hydrolysis indicate that urea was almost completely hydrolyzed within a few hours of urine mixing with feces. The estimated urea hydrolysis rates from this study are likely maximum rates because of the thorough mixing before each sampling. Although considerable mixing of feces and urine occurs on the barn floor of commercial dairy operations from cattle walking through the manure, such mixing may be not as quick and thorough as in this study. Consequently, the urea hydrolysis rates from this study indicate the maximum loss of urea and should be accounted for in management aimed at mitigating ammonia emissions from dairy cattle manure under similar urea concentration, pH, and temperature conditions reported in this experiment.