sIgA Concentration Research Articles

Emotional responses of piglets under long-term exposure to negative and positive auditory stimuli

The stress caused by sound is inevitable. The stress caused by noise and the positive effects of music can affect the endocrine of animals and their welfare. In this study, a total of 72 hybrid piglets (Large White × Duroc × Min pig) were randomly divided into 3 groups, including music (Mozart K.448, 60–70 dB), noise (recorded mechanical noise, 80-85 dB), and control (natural background sound, <40 dB) groups. S-IgA (secretory immunoglobulin A), IL-6 (interleukin-6), IL-8 (interleukin-8), and positive emotion-related behaviors were used as indicators to discuss whether noise induced stress and inflammation in piglets or whether music could have positive effects. Six hours of auditory exposure were given daily (10:00–16:00), which lasted for 56 days. Behavioral responses of the piglets were observed, and the concentrations of salivary S-IgA and serum IL-6 and IL-8 were measured. The results showed that the concentration of S-IgA increased in the noise and control groups on the 57th day (P < 0.05); S-IgA concentration in the music group was unchanged after long-term music exposure. The concentrations of IL-6 and IL-8 showed that long-term noise exposure might lead to stress and inflammation in piglets. Tail-wagging and play behaviors of the piglets in the music group were significantly greater than those in the noise and control groups, which implied that long-term music exposure improved the emotional state of the piglets in a restricted and barren environment.

Comparison of Effects of Storage at Different Temperatures in a Refrigerator, Upright Freezer on Top of Refrigerator, and Deep Freezer on the Immunoglobulin A Concentration and Lysozyme Activity of Human Milk.

This study aimed to investigate the effects of storing expressed human milk (HM) at different domestic storage temperatures on the secretory immunoglobulin A (SIgA) concentration and lysozyme activity. Forty mothers of full-term infants aged one to six months provided milk samples. The fresh samples were examined within 24 h of expression, and the other samples were stored in a refrigerator for four days or in two types of freezers for six months. The SIgA concentrations and lysozyme activity in the milk samples were studied using enzyme-linked immunosorbent assay (ELISA) kits and fluorometric lysozyme activity assay kits, respectively. The pairwise comparisons of the SIgA concentration and lysozyme activity were carried out using one-way analysis of variance with Dunnett T3 or Kruskal–Wallis tests with Bonferroni correction, depending on the data distribution. The mean temperatures of the refrigerator, upright freezer on top of the refrigerator, and deep freezer (chest freezer) were 2.0, −16.7, and −22.3 °C, respectively. Our study results highlight that the SIgA concentration and lysozyme activity of HM stored in the refrigerator for four days and in freezers for six months were significantly lower than those of fresh HM (p < 0.001). During the first six months of storage in both types of freezers, the SIgA levels were stable, whereas the lysozyme activity significantly decreased (p < 0.001). HM stored in the deep freezer had a higher SIgA concentration and lysozyme activity than HM stored in the upright freezer on top of the refrigerator. Our data support the superiority of fresh human milk over stored HM. If HM is to be stored, then storage in a deep freezer is potentially a more effective method for the preservation of SIgA concentrations and lysozyme activity than storage by refrigeration for four days or in an upright freezer on top of a refrigerator for six months.

Mucosal immunity during rehabilitation in the patients after coronavirus infection

Following the coronavirus infection, a variety of symptoms may persist for a long time. Therefore, an important area of further research is to study the state and response of protective immune mechanisms in the post-COVID period. Our aim was to evaluate the dynamics of mucosal immunity by measuring sIgA in the saliva samples and nasal swabs (sIgA, lactoferrin), as well as efficiency of interferon-alpha-2b (IFNa2b) treatment in the patients after coronavirus infection.&#x0D; A study was conducted in the patients aged 18 to 60 years (n = 130) at the terms of 1 to 9 months after a coronavirus infection. A control group consisted of conditionally healthy individuals (n = 15). Diagnosis of post-COVID manifestations was carried out by collecting complaints, anamnestic data, physical examination and questionnaire. The state of mucosal immunity in saliva samples and nasopharyngeal mucosal scrapings was evaluated in dynamics as based on determination of sIgA and lactoferrin concentrations by means of enzyme immunoassay techniques prior to administration of local preventive therapy with recombinant IFNa2b, and 1 month after the treatment (VIFERON gel applied intranasally twice a day for 30 days).&#x0D; The following symptoms of post-COVID syndrome were documented In the study group: joint and muscular pain, shortness of breath, cough, fatigue and weakness, headache and dizziness, anxiety. In the group of patients who received preventive therapy during 1 to 3 months after coronavirus infection, a significantly increased level of saliva sIgA was noted, respectively, 1.840.28 to 5.781.96 mg/mL. As based on the data obtained with scrapings from nasopharyngeal mucosa, a significant increase in the level of sIgA was revealed in the group subjected to therapy up to 3 months after COVID-19 infection, i.e., from 28.613.0 to 39.833.85 mg/mL. In the group of patients devoid of preventive therapy, a stable maintenance of the reduced mucosal immunity parameters was found in all time intervals during the period of convalescence. In all observed patients, regardless of the group, a decreased lactoferrin level was found, being two-fold lower than the normal reference values. The incidence of respiratory viral infections in the group without preventive therapy was statistically significant, being registered in 9.2% of cases.&#x0D; Patients after COVID-19 infection exhibit a persistent decrease in mucosal immunity. Immunological efficacy was observed when using IFNa2b, thus making it possible to recommend it for rehabilitation in this group of patients over the period of convalescence.

Effects of lactic acid bacteria-fermented formula milk supplementation on ileal microbiota, transcriptomic profile, and mucosal immunity in weaned piglets

BackgroundLactic acid bacteria (LAB) participating in milk fermentation naturally release and enrich the fermented dairy product with a broad range of bioactive metabolites, which has numerous roles in the intestinal health-promoting of the consumer. However, information is lacking regarding the application prospect of LAB fermented milk in the animal industry. This study investigated the effects of lactic acid bacteria-fermented formula milk (LFM) on the growth performance, intestinal immunity, microbiota composition, and transcriptomic responses in weaned piglets. A total of 24 male weaned piglets were randomly divided into the control (CON) and LFM groups. Each group consisted of 6 replicates (cages) with 2 piglets per cage. Each piglet in the LFM group were supplemented with 80 mL LFM three times a day, while the CON group was treated with the same amount of drinking water.ResultsLFM significantly increased the average daily gain of piglets over the entire 14 d (P < 0.01) and the average daily feed intake from 7 to 14 d (P < 0.05). Compared to the CON group, ileal goblet cell count, villus-crypt ratio, sIgA, and lactate concentrations in the LFM group were significantly increased (P < 0.05). Transcriptomic analysis of ileal mucosa identified 487 differentially expressed genes (DEGs) between two groups. Especially, DEGs involved in the intestinal immune network for IgA production pathways, such as polymeric immunoglobulin receptor (PIGR), were significantly up-regulated (P < 0.01) by LFM supplementation. Moreover, trefoil factor 2 (TFF2) in the LFM group, one of the DEGs involved in the secretory function of goblet cells, was also significantly up-regulated (P < 0.01). Sequencing of the 16S rRNA gene of microbiota demonstrated that LFM led to selective enrichment of lactate-producing and short-chain fatty acid (SCFA)-producing bacteria in the ileum, such as an increase in the relative abundance of Enterococcus (P = 0.09) and Acetitomaculum (P < 0.05).ConclusionsLFM can improve intestinal health and immune tolerance, thus enhancing the growth performance of weaned piglets. The changes in microbiota and metabolites induced by LFM might mediate the regulation of the secretory function of goblet cells.

IgA-Based Secretory Response in Tears of COVID-19 Patients: A Potential Biomarker of Pro-Inflammatory State in Course of SARS-CoV-2 Infection.

Mucosal immunity, including secretory IgA (sIgA), plays an important role in the early defence against SARS-CoV-2 infection. However, a comprehensive evaluation of the local immune response in tears in relation to blood antibody reservoirs has not yet been conducted. A total of 179 symptomatic laboratory-confirmed COVID-19 patients were included in this single-centre study. Conjunctival swabs were analysed by a reverse transcription polymerase chain reaction for the detection of SARS-CoV-2 RNA. In parallel, tear samples collected by Schirmer test strips and plasma samples were analysed by ELISA to detect anti-S1 IgA levels. The concentrations of selected inflammatory cytokines in tears were determined by a magnetic bead assay. Anti-SARS-CoV-2 sIgA was present in the tears of 81 (45.25%) confirmed COVID-19 patients, and the tear IgA levels were correlated with the plasma IgA levels (Rs = +0.29, p = 0.0003). SARS-CoV-2 RNA in the conjunctival sac was identified in 18 COVID-19 patients (10%). Positive correlations between the tear IgA level and the concentrations of several cytokines TNF-α (Rs = +0.23, p = 0.002), IL-1β (Rs = +0.25, p < 0.001), IL-2 (Rs = +0.20, p = 0.007), IL-4 (Rs = +0.16, p = 0.04), IL-5 (Rs = +0.36, p < 0.001), IL-6 (Rs = +0.32, p < 0.001), IL-8 (Rs = +0.31, p < 0.001), VEGF (Rs = +0.25, p < 0.001) and GM-CSF (Rs = +0.27, p < 0.001) were also found. Quantitative tear film-based sIgA could potentially serve as a rapid and easily accessible biomarker of external mucosal immunity to SARS-CoV-2. The concentration of sIgA is directly related to individual host immune responses to SARS-CoV-2 infection.

Changes in immunological parameters and umbilical blood flow in pregnant women during exacerbation of chronic bronchitis associated with cytomegalovirus infection

Aim. To evaluate changes in immunological parameters and umbilical blood flow in pregnant women with exacerbation of chronic bronchitis associated with cytomegalovirus infection (CMVI).Materials and methods. The concentration of secretory immunoglobulin A (sIgA), immunoglobulin E (IgE), circulating immune complexes (CIC), interleukin- 6 (IL-6), interleukin-4 (IL-4) and systolic-diastolic ratio (SDR) in the umbilical artery was studied in 80 women with exacerbation of chronic bronchitis in the second trimester of pregnancy, uncomplicated and complicated by the acute phase of CMVI. The first group was represented by 30 women with cytomegalovirus seronegative physiological pregnancy; the second – 25 patients with exacerbation of chronic bronchitis caused by reactivation of CMVI, complicated by the development of chronic compensated placental insufficiency (CCPI); the third – 25 women with chronic simple bronchitis in the acute stage in the setting of the acute phase of CMVI, initiating the formation of chronic subcompensated placental insufficiency (CSPI) at 30-34 weeks of gestation.Results. In the first group, the concentration of sIgA in blood serum was 4.3±0.26 mg/L, IgE – 3.64±0.23 IU/mL, CIC – 0.091±0.005 AU, IL-6 – 2.41±0.21 pg/mL, IL-4 – 30.1±2.19 pg/mL, cortisol – 527.2±20.95 nmol/L, and SDR in the artery umbilical cord – 3.41±0.06 relative units. In women of the second group, in comparison with the first one, there was an increase in the concentration of sIgA by 1.40 times (p&lt;0.01), IgE – by 1.95 times (p&lt;0.001), CIC – by 1.83 times (p&lt;0.001) , IL-6 – 2.61 times (p&lt;0.001), IL-4 – 1.29 times (p&lt;0.05), cortisol – 1.35 times (p&lt;0.001) in the absence of statistically significant differences the magnitude of vascular resistance in the umbilical artery. In the third group, in comparison with the first one, there was a decrease in sIgA values by 1.26 times (p&lt;0.05) in the setting of an increase in the concentration of IgE by 2.56 times (p&lt;0.001), CIC – by 2.09 times (p&lt;0.001 ), IL-6 – by 3.65 times (p&lt;0.001), IL-4 – by 1.64 times (p&lt;0.001), cortisol – by 1.52 times (p&lt;0.001) and SDR in the umbilical artery – 1.27 times (p&lt;0.001). In women of the third group, in contrast to the second one, a decrease in the concentration of sIgA by 1.77 times (p&lt;0.001), as well as an increase in IgE by 1.31 times (p&lt;0.01), CIC ‒ by 1.14 times (p&lt;0.001), IL-6 ‒ 1.39 times (p&lt;0.001), IL-4 ‒ 1.27 times (p&lt;0.05), cortisol ‒ 1.13 times (p&lt;0.05) and SDR in the umbilical artery ‒ 1.21 times (p&lt;0.001). A direct correlation was established between IL-6 and cord blood flow (r=0.53; p&lt;0.01), as well as an inverse relationship between sIgA and cortisol (r=-0.44; p&lt;0.05).Conclusion. In the second trimester of gestation, exacerbation of chronic simple bronchitis caused by reactivation of CMVI, which initiates the development of CSPI, in contrast to chronic bronchitis in the acute stage, associated with the acute phase of chronic CMVI, which induces the formation of CCPI, is accompanied by an imbalance of nonspecific local and systemic immune responses, as well as stress-reaction, which leads to the growth of SDR in the umbilical artery.

Biomarkers for monitoring the equine large intestinal inflammatory response to stress-induced dysbiosis and probiotic supplementation.

Large intestine barrier disturbances can have serious consequences for the health of horses. The loss of mucosal integrity that leads to increased intestinal permeability may result from a local inflammatory immune response following alterations of the microbiota, known as dysbiosis. Therefore, our research aimed to identify noninvasive biomarkers for studying the intestinal permeability and the local inflammatory immune response in horses. Regarding the biomarkers used in other mammalian species, we measured the concentrations of lipopolysaccharides (LPS), reflected by 3-OH C14, C16, and C18 fatty acids, in blood, and fecal secretory immunoglobulin-A (SIgA). These biomarkers were evaluated in two trials including 9 and 12 healthy horses, which developed large intestinal dysbiosis experimentally induced by 5 d of antibiotic administration (trimethoprim sulfadiazine [TMS]) or 5 d of abrupt introduction of high starch levels (barley) into the diet. Horses were either control or supplemented with Lactobacillus acidophilus, Ligilactobacillus salivarius, and Bifidobacterium lactis. Correlations were performed between biomarkers and fecal bacterial diversity, composition, and function. No significant interaction between day and supplementation, or supplementation effect were observed for each biomarker. However, with the dietary stressor, a significant increase in blood concentrations of 3-OH C16 (P = 0.0125) and C14 (P = 0.0252) fatty acids was measured 2 d after the cessation of barley administration. Furthermore, with the antibiotic stressor, blood levels of 3-OH C16 progressively increased (P = 0.0114) from the first day to 2 d after the end of TMS administration. No significant day effect was observed for fecal SIgA concentrations for both stressors. These results indicate that both antibiotic- and diet-induced dysbiosis resulted in a local translocation of LPS 2 d after the cessation of the stressor treatments, suggesting an impairment of intestinal permeability, without detectable local inflammation. Blood LPS and fecal SIgA concentrations were significantly correlated with several bacterial variations in the large intestine, which are features of antibiotic- and diet-induced dysbiosis. These findings support the hypothesis that a relationship exists between dysbiosis and the loss of mucosal integrity in the large intestine of horses.

Biochemical and immunological markers of the adaptive response in olfacto-odorimetric studies

The purpose of the study is to determine whether exposure to odours of model food odourants can lead to a change in biochemical and immunological parameters that we previously used when examining the population in the area of food industry enterprises location using the method of quantitative olfacto-odorimetry. Methods. The specified concentrations of aerosols of three food flavours (orange, cognac and coffee) were supplied to the participants of the studies with a help of ECOMA T08 olfactometer. Quantitative composition of the aerosols was controlled by GC/MS. In participants saliva samples taken before, during and at the end of each experiment, the intensity of luminol-dependent chemiluminescence, the content of secretory IgA, IL-1β, IL-6 and IL-8, the activity of α-amylase and N-acetyl-β-D-glucosaminidase were determined. For data analysis, paired Friedman and Wilcoxon tests were used with Bonferroni correction for the problem of multiple comparisons. Results. A reliable effect of the smell of food odourants was found on one indicator only - the activity of salivary α-amylase - when combining data from 5 separate experiments (n=45): 93.3[24.3;160.0] U/ml at the end of the experiments against background values of 109.9 [42.5; 216.7] U/ml;, p=0.0096 with a significance level of p=0.05/3=0.017. A decrease in the average values of salivary α-amylase activity was shown to hide opposite changes in individual values: an increase in activity in people with low background values (below the median of the initial distribution) and an amplitude-dominant decrease - in people with high background values (above median). The revealed phenotypic polymorphism of α-amylase regulation contributes to one of relevant Post-COVID areas - the study of the ability of people to perceive odours and react to them. Limitations. The use of olfacto-odorimetry to study effect of odours on human health indicators is promising, but requires design of protocols with extended exposure time. Conclusion. A decrease in average values of salivary α-amylase activity with distinctive forestall of the upper quartile may be a sign of human reflex re sponse to the emission of odourous substances in the areas of food industry.

Heat acclimation does not negatively affect salivary immunoglobulin-A and self-reported illness symptoms and wellness in recreational athletes

ABSTRACT Heat acclimation (HA) protocols repeatedly expose individuals to heat stress. As HA is typically performed close to the pinnacle event, it is essential that the protocol does not compromise immune status, health, or wellbeing. The purpose of this study was to examine the effect of HA on resting salivary immunoglobulin-A (s-IgA) and salivary cortisol (s-cortisol), self-reported upper-respiratory tract symptoms, and self-reported wellness parameters. Seventeen participants (peak oxygen uptake 53.2 ± 9.0 mL·kg−1·min−1) completed a 10-day controlled-hyperthermia HA protocol, and a heat stress test both before (HST1) and after (HST2) HA (33°C, 65% relative humidity). Resting saliva samples were collected at HST1, day 3 and 7 of the HA protocol, HST2, and at 5 ± 1 days post-HA. Upper-respiratory tract symptom data were collected weekly from one week prior to HA until three weeks post HA, and wellness ratings were reported daily throughout HA. HA successfully induced physiological adaptations, with a lower end-exercise rectal temperature and heart rate and higher whole-body sweat rate at HST2 compared to HST1. In contrast, resting saliva flow rate, s-IgA concentration, s-cortisol concentration, and s-cortisol secretion rate remained unchanged (n = 11–14, P = 0.10–0.48). Resting s-IgA secretion rate increased by 39% from HST1 to HST2 (n = 14, P = 0.03). No changes were observed in self-reported upper respiratory tract symptoms and wellness ratings. In conclusion, controlled-hyperthermia HA did not negatively affect resting s-IgA and s-cortisol, self-reported upper-respiratory tract symptoms, and self-reported wellness parameters in recreational athletes.

Evaluation of Intestinal Microbial Metabolites in Preterm Infants with Different Initial Feeding Methods by In Vitro Fermentation Modeling System.

We aim to explore the intestinal microbial metabolites in preterm infants with noninvasive methods and analyze the effects of initial feeding methods. Preterm infants with gestational weeks lower than 34 were recruited for fecal sample collection every 7 days. Fecal pH, ammonia, bile acid, and secretory IgA (sIgA) were tested. A 1:10 fecal slurry was inoculated into different culture media containing different carbohydrates as the only carbon source: lactose (LAT), fructooligosaccharide (FOS), galactooligosaccharide (GOS), and 2′-fucosyllactose (FL2). After 24 h of anaerobic culture through an in vitro fermentation system, air pressure difference, carbohydrate degradation rate, and short-chain fatty acids (SCFAs) content in fermentation pots were measured. Preterm infants were assigned into two groups: group A, preterm infants fed by human milk, including mother’s own milk and donor human milk (DHM); group B, preterm infants fed by preterm formula at first 3 days and fed by human milk (including mother’s own milk and DHM) from day 4 to discharge. Group A included 90 samples and group B included 70 samples. Group A had lower fecal pH (p = 0.023), ammonia (p = 0.001), and bile acids (p = 0.025). Group B also had higher fecal sIgA levels, both in OD (p = 0.046) and concentration (p < 0.0001) methods. Carbohydrates degradation rates in group A were higher than group B, especially in LAT medium (p = 0.017) and GOS medium (p = 0.005). Gas production amount had no significant difference in all four media. Several different SCFAs in four kinds of different culture media in group A were higher than in group B, but valeric acid was lower in group A. The initial feeding methods may affect the preterm infants’ intestinal microecology and microbial metabolites for at least several weeks.

Effects of Housing System on Anxiety, Chronic Stress, Fear, and Immune Function in Bovan Brown Laying Hens.

Simple SummaryThe objectives of this study were to determine if housing Bovan brown laying hens in conventional cages or enriched floor pens impacted novel physiological and behavioral markers for animal welfare and whether we can use these markers to assess animal welfare. We found that birds that were housed in conventional cages showed increased tonic immobility durations (indication of fearfulness), decreased fecal Immunoglobulin A (indicator of immune function), and increased feather corticosterone concentrations (indicator of chronic stress) compared to hens that were housed in enriched pens. These results indicate that caged birds are more stressed, have reduced immune function, and are more fearful than birds that are housed in pens. In contrast to expectations, we found that caged hens showed a shorter latency to feed during attention bias testing, indicating reduced anxiety compared to birds from pens. Overall, we found that conventional cages generally impacted animal welfare negatively, with the exception of anxiety. In addition, the results suggest that the chosen novel markers for animal welfare show appropriate contrast between long-term housing systems for laying hens. Yet, additional work needs to be done before these measures can be used more broadly.The scientific community needs objective measures to appropriately assess animal welfare. The study objective was to assess the impact of housing system on novel physiological and behavioral measurements of animal welfare for laying hens, including secretory and plasma Immunoglobulin (IgA; immune function), feather corticosterone (chronic stress), and attention bias testing (ABT; anxiety), in addition to the well-validated tonic immobility test (TI; fearfulness). To test this, 184 Bovan brown hens were housed in 28 conventional cages (3 birds/cage) and 4 enriched pens (25 birds/pen). Feces, blood, and feathers were collected 4 times between week 22 and 43 to quantify secretory and plasma IgA and feather corticosterone concentrations. TI tests and ABT were performed once. Hens that were from cages tended to show longer TI, had increased feather corticosterone, and decreased secretory IgA at 22 weeks of age. The caged hens fed quicker, and more hens fed during the ABT compared to the penned hens. Hens that were in conventional cages showed somewhat poorer welfare outcomes than the hens in enriched pens, as indicated by increased chronic stress, decreased immune function at 22 weeks of age but no other ages, somewhat increased fear, but reduced anxiety. Overall, these novel markers show some appropriate contrast between housing treatments and may be useful in an animal welfare assessment context for laying hens. More research is needed to confirm these findings.

Effects of the thawing rate and heating temperature on immunoglobulin A and lysozyme activity in human milk

BackgroundThe percentage of infants receiving frozen human milk (HM) is increasing. The effects of thawing and warming on the secretory immunoglobulin A (SIgA) level and lysozyme activity in frozen HM should be investigated to identify optimal methods for preserving immune factors in frozen HM.MethodsMilk samples were collected from 40 mothers with healthy full-term infants who had been lactating for one to six months. The baseline samples were analyzed within 24 h after collection, and the other samples were frozen at -18 °C before analyses. We compared two methods: placing the container overnight in a refrigerator at 4 °C before warming (slow thawing) and immediately thawing in warm water after removing the sample from the freezer (rapid thawing). Additionally, we investigated the effects of the warming temperature by comparing room temperature (25 °C) and physiological temperature (37 °C). The SIgA concentrations and lysozyme activities in the milk samples were determined using ELISA kits and fluorometric lysozyme activity assay kits, respectively.ResultsThe SIgA concentrations and lysozyme activity in frozen HM were 16.5–52.1% and 16.8–39.3% lower than those in fresh HM, respectively. The SIgA concentrations in frozen HM were stable during slow thawing at 37 °C (p = 0.072) compared with those in fresh HM. The SIgA concentrations and lysozyme activity were maintained at significantly higher levels during slow thawing than during rapid thawing at 25 °C (p = 0.002 and p < 0.001, respectively). Slow thawing preserved higher SIgA concentrations and lysozyme activity than rapid thawing at 37 °C, but the difference was not significant.ConclusionsThe SIgA level in HM frozen at -18 °C for two months was stable after overnight thawing in the refrigerator (4 °C for 12 h) before warming to 37 °C compared with that in fresh milk. The thawing of HM in the refrigerator overnight (and then warming to 25 °C or 37 °C for 30 min) has the potential to preserve the SIgA concentration and lysozyme activity to a greater extent than heating immediately after removal from the freezer. Broader temperature ranges should be analyzed to determine the temperature that minimizes the losses in SIgA concentration and lysozyme activity in HM.

An impact of lactoferrin, serum albumin and secretory immunoglobulin A in actimicrobial activity of breast milk whey

The contribution of the antimicrobial activity of sIgA, lactoferrin, -lactalbumin, serum albumin, and lysozyme to the total antimicrobial activity of whey was evaluated. 66 breast milk samples from healthy women aged from 23 to 45 years collected at different periods of lactation were studied. It was found that along with proceeding the lactation period, not only the total antimicrobial activity (r = 0.944) decreased, but also the activity of the low molecular weight fraction containing the sum of antimicrobial peptides (r = 0.950). The median lactoferrin level varied from 3.46 mg/ml in colostrum down to 0.94 mg/ml one year after the onset of lactation that correlated with whey activity (r = 0.616). The concentration of sIgA was peaked in the whey of colostrum (5.01 mg/ml) and significantly reduced in mature milk, remaining approximately at the same level in subsequent periods of lactation (1.0 mg/ml). Serum albumin concentration significantly decreased while proceeding through lactation period (from 5.52 to 4.68 mg/ml) and correlated with whey activity (r = 0.589). Evaluating effects of the purified -lactalbumin on C. albicans cells within the range of 520 mg/ml by spectrophotometry and microscopy showed the absence of fungicidal activity. At the same time, purified lactoferrin, IgA, lysozyme, and serum albumin demonstrated a direct fungicidal effect. The activity of the IgA preparation at a concentration relevant to the sIgA content in the whey of the first day colostrum was 50.0%, and in the whey after 12 months 31.0%; lactoferrin 26.7% and 3.4%; serum albumin 15.0% and 17.7%; lysozyme 0.1% and 1.8%. Thus, the sum of the activities of these polypeptides at concentrations typical for the first-day colostrum was 91.7%, which is comparable to the total activity of this whey 82.4%. The sum of the activities of these polypeptides, typical for 12 months of lactation, was 53.9%, and the total whey activity relevant to this period was 64.5%. Therefore, IgA and lactoferrin contribute the most significant antimicrobial activity in the whey of colostrum, whereas after 12 month-breastfeeding IgA and serum albumin become most significant.

A Randomized Controlled Trial of Dietary Rice Bran Intake on Microbiota Diversity, Enteric Dysfunction, and Fecal Secretory IgA in Malian and Nicaraguan Infants

ABSTRACTBackgroundMalnutrition and diarrhea are leading causes of death in children aged <5 y. Rice bran is a nutrient-dense prebiotic available globally.ObjectivesThe objective of this secondary analysis was to evaluate the effects of daily rice bran supplementation on environmental enteric dysfunction (EED) markers, total fecal secretory IgA (sIgA), and microbiota in infants at high risk of malnutrition.MethodsSix-month-old Malian and Nicaraguan infants were randomly assigned to control or daily rice bran supplementation cohorts (1 to 5 g/d). Feces were collected monthly for 6 mo to evaluate fecal sIgA, markers of EED, and microbiota diversity. Statistical methods included linear mixed models, generalized mixed models, Spearman correlation, and Wilcoxon rank-sum tests.ResultsSix-month-old Malian infants had significantly elevated sIgA (4.0× higher, P < 0.001), fecal myeloperoxidase (31.6× higher, P < 0.001), fecal α1-antitrypsin (1.8× higher, P = 0.006), and lower fecal neopterin (0.13× higher, P < 0.001) than the age-matched Nicaraguan infants. In the Nicaraguan rice bran cohort from 6 to 12 mo of age, there was a significant decrease in sIgA concentrations (0.4×, P < 0.05) and a correlation between sIgA and the EED marker α1-antitrypsin (0.523, P < 0.0001) at 12 mo of age. In Malian infants, daily rice bran ingestion resulted in decreased EED scores (0.71×, P = 0.02) and a stable sIgA concentration over time. The rice bran group of Malian infants also had correlation between sIgA and the EED marker neopterin (0.544, P < 0.001) at 12 mo of age and a significant (P < 0.05) increase in microbiota α-diversity at a younger age (9 mo with rice bran compared with 10 mo in control group), which supports earlier microbiota maturation.ConclusionsThese results support rice bran as a functional food ingredient targeting gut mucosa in children at high-risk of malnutrition.

DYNAMICS OF SIgA CONTENT IN BREAST MILK WHEN RISK FACTORS AND LACTATIONAL CHARACTERISTICS ARE CONSIDERED

The leading role associated with an anti-infective action of breast milk belongs to secretory IgA (SIgA). Therefore, the determination of the level of SIgA in colostrum and milk of mothers with different lactation levels and mothers at risk at different levels and duration of lactation has a practical interest.&#x0D; Aim. The research aims at studying the dynamics of SIgA content in breast milk, taking into account risk factors and features of lactation.&#x0D; Material and Methods. The content of SIgA in colostrum and breast milk of 372 mothers with full lactation and 208 with hypogalactia, of which 72 with early and 146 with late hypogalactia at different times of lactation (1-8 days and 1-3 months) was determined. The SIgA content was also studied in mothers; 65 with preeclampsia, including 12 with early hypogalactia, 24 with late hypogalactia, 23 with full lactation, and 44 with anemia of pregnancy, including 12 with early hypogalactia, 19 with late hypogalactia and 13 with full lactation. The determination of SIgA content in colostrum and milk was performed using the method of simple radial immunodiffusion in a gel by G. Mancini et al.&#x0D; Results and Discussion. The SIgA concentration in colostrum and breast milk decreased during the secretion process. In early hypogalactia, the content of SIgA did not differ significantly from the level of SIgA in colostrum and breast milk with full lactation in the early neonatal period and at 1-3 months of secretion. In the late hypogalactia, SIgA content did not differ significantly from its level in the control group. Due to the lower amount of milk received by the mother's child with hypogalactia, the supply of children with SIgA is insufficient, and the deficit is higher the sooner the hypogalactia develops. Similar changes in the dynamics of SIgA content have been found in mothers with preeclampsia and at different levels of lactation. The exception was the content of SIgA in colostrum and milk of mothers with anemia of pregnancy and early hypogalactia, which in the first five days of lactation was lower than in mothers with full lactation.&#x0D; Conclusion. The SIgA concentration in colostrum and breast milk decreases during the secretion process. The level of lactation does not affect the dynamics of SIgA content, except for the SIgA content in colostrum and breast milk of mothers with anemia of pregnancy and hypogalactia in the early stages of secretion. Despite the absence of a significant difference in SIgA levels in mothers with different levels of lactation due to less colostrum and milk received by children with early and late hypogalactia, the supply of SIgA is insufficient, and the cumulative deficit is higher the faster the hypogalactia develops.

Research note: effects of different anticoccidial regimens on the growth performance, hematological parameters, immune response, and intestinal coccidial lesion scores of yellow-feathered broilers

Yellow-feathered broiler chickens generally have a longer growth cycle than white broilers and therefore face different coccidiosis challenges. General single vaccine and drug regimens struggle to prevent coccidiosis for yellow broilers. In this study, a single vaccine, and a combination of coccidiostat regimens was employed to explore the preventative and control effects of different pilot regimens on coccidiosis in yellow-feathered broilers. A total of 2,000 one day old Chinese Huang Youma female broilers were allocated into 4 experimental groups, each with 5 replicates. All birds were fed the same starter feed from Days 1 to 25, and all groups were inoculated with a vaccine on Day 4. After Day 26, the groups were then fed as follows: (1) Negative control group: basal diet + vaccine (NC); (2) NC + maduramycin (NCMD); (3) NC + narasin (NCNR); and (4) NC + salinomycin (NCSL). From Days 26 to 75, the NCNR group had a lower FCR than the other groups. The 75-d BW was higher in the NCNR group than in the NCSL group but was not significantly higher than that in the NC and NCMD groups. The growth performance followed the same trend during the whole experiment (Days 1–75). Compared to the NC group, the NCNR and NCSL groups had higher intestinal mucosa SIgA concentrations at Day 40 and Day 60 (P < 0.001); however the NCMD group had lower IgG levels at Day 40 and Day 60 (P = 0.036, P = 0.006 respectively). The combination groups had significantly reduced AKP levels and urine acid concentrations at Day 60 in comparison to those of the NC group (P = 0.004). The NCNR and NCMD groups had less severe intestinal coccidiosis lesion scores than the NC and NCSL groups in older birds. Thus, a single vaccine and/or combinations with different coccidiostats had different effects on broilers. The NCNR group showed comparatively better growth performance, blood biochemical indices, immune response, and coccidiosis lesion scores.

Early Weaning Stress Induces Intestinal Microbiota Disturbance, Mucosal Barrier Dysfunction and Inflammation Response Activation in Pigeon Squabs.

Early weaning stress has been reported to impair intestinal health in mammals. Like mammals, weaning of the pigeon squab, an altricial bird, is associated with social, environmental and dietary stress. However, understanding of weaning stress on intestinal functions is very limited in altricial birds, especially in squabs. This study was aimed to evaluate the effects of early weaning stress on intestinal microbiota diversity, architecture, permeability, the first line defense mechanisms, mucosal barrier functions, and immune cell responses. A total of 192 newly hatched squabs were randomly allocated into two groups, one weaned on day 7 and the other remained with the parent pigeons. Mucosal tissue and digesta in ileum, as well as blood samples, were collected from squabs (n = 8) on days 1, 4, 7, 10, and 14 postweaning. Our results showed that weaning stress induced immediate and long-term deleterious effects on both growth performance and intestinal barrier functions of squabs. Early weaning significantly increased ileal bacterial diversity and alters the relative abundance of several bacteria taxa. Weaning stress can also cause morphological and functional changes in ileum, including an atrophy in villi, an increase in permeability, and a variation in the mRNA expression of genes encoding mucins, immunoglobulins, tight junction proteins, toll-like receptors, and cytokines, as well as the concentration of secretory IgA. We concluded that the impaired intestinal barrier functions accompanied with early weaning stress seems to be the main reason for the poor growth rate after weaning in squabs. In addition, the disturbance of intestinal microbiota of early weaning stress in squabs coincided with dysfunction of intestinal mucosal barrier and activation of inflammation cell responses that were possibly mediated via the activation of toll-like receptors.

Особенности иммунитета и возможности коррекции дезадаптации иммунного ответа у больных острыми респираторными вирусными инфекциями из организованных коллективов

The purpose of the study was to determine the severity of maladaptation disorders of the immune response and the possibility of their correction in patients with acute respiratory viral infections from organized groups. Material and methods. 90 patients with uncomplicated forms of ARVI and a group of conditionally healthy (n = 30) from organized groups were examined. In addition to symptomatic therapy, patients received a combination of rectal and intranasal forms of recombinant IFN-α-2b. Identification of the ARVI pathogen was determined by PCR. ELISA was used to determine serum concentrations of IFN-α and IFN-γ and the ability of blood cells to produce these cytokines ex vivo spontaneously and when stimulated by Newcastle disease virus or phytohemagglutinin. Flow cytometry was used to analysis the subpopulation composition of peripheral blood lymphocytes. The study of the levels of secretory IL-8, IgA, serum IL-8 was carried out by enzyme immunoassay. Results. The etiology of ARVI was deciphered in 62.1% of patients, the main share in the morbidity structure was due to influenza (39.3%) and adenoviruses (35.6%). In the first 3 months of the formation of organized teams in apparently healthy people, a decrease in the level of T-helpers, natural killers and inhibition of the interferon system was revealed, which indicates a violation of adaptive mechanisms and contributes to a decrease in body resistance. An analysis of the cytokine system showed an increase in the concentration of secretory and serum IL-8 in patients with acute respiratory viral infections compared with a group of practically healthy people. In the immune status 1 month after ARVI, there was a significant increase in the number of T-lymphocytes, T-helpers with a tendency to normalize T-lymphocytes carrying IFN-α and -γ receptors, while maintaining a significantly reduced number of B-lymphocytes. The immunomodulatory efficacy of combination therapy with recombinant IFN-α-2b is characterized by an increase in the amount of induced IFN-α during its initial depression and a decrease in initial hyperproduction. Also, at the initial level of induced IFN-α less than 200 pg/ml, there was an increase in the concentration of secretory IL-8 and IgA. Conclusion. In conditionally healthy individuals, in the first 3 months of the formation of organized groups, maladjustment disorders in the immune response were detected in 34% of cases. The interferon-modulating effect of combination therapy with recombinant IFN-α-2b was shown.

Construction of the waaF Subunit and DNA Vaccine Against Escherichia coli in Cow Mastitis and Preliminary Study on Their Immunogenicity.

Escherichia coli (E. coli) is one of the major pathogenic bacteria in bovine mastitis, which usually triggers systemic symptoms by releasing lipopolysaccharide (LPS). waaF is the core in LPS pathogenicity. In this study, a new waaF vaccine candidate was identified, constructed with the pcDNA3.1 (+)HisB-waaF plasmid to create to a DNA vaccine (pcwaaF), and transfected into MCF-7 cells to produce recombinant waaF subunit vaccine (rwaaF). After that, the safety of the two vaccine candidates was evaluated in mouse model. Immunogenicity and mortality of challenged mice were compared in 20 and 40 μg per dose, respectively. The results showed that rwaaF and pcwaaF were successfully constructed and the complete blood count and serum biochemical indicated that both of the vaccine candidates were safe (p > 0.05). In addition, histopathological staining showed no obvious pathological changes. The immune response induced by rwaaF was significantly higher than that of pcwaaF (p < 0.01), indicated by levels of serum concentration of IgG IL-2, IL-4, and IFN-γ, and feces concentration of sIgA. Survival rates of mice in rwaaF groups (both 80%) were also higher than in the pcwaaF groups (40 and 50%, respectively). Comparing the safety, immunogenicity, and E. coli challenge of two vaccine candidates, rwaaF had the better effect and 20 μg rwaaF was more economical. In conclusion, this study demonstrates the utility of a new E. coli vaccine and provides a rationale for further investigation of bovine mastitis therapy and management.

Total Protein, Secretory Immunoglobulin A and Lactoferrin Concentrations in the Breastmilk of Lactating Women and their Correlation with Nutritional Intake

Background and objectives: Mothers’ breastmilk is the best source of nutrition for infants; hence, its nutritional adequacy is essential. The purpose of this study is to evaluate the total protein, secretory immunoglobulin A (sIgA) and lactoferrin (LF) concentrations of breastmilk from lactating mothers in Jakarta and analyse their correlation with average daily calory, macronutrient and vitamin C and E intakes. Methods and study design: A cross-sectional study was conducted in two primary health centres in Jakarta, Indonesia. The breastmilk’s total protein concentration was measured with the Warburg–Christian method, while sIgA and LF concentrations were measured by enzyme-linked immunosorbent assay (ELISA). Macronutrient and vitamin C and E intake was assessed through 24-hour food recall and a semi-quantitative food frequency questionnaire (FFQ), respectively. Results: Thirty-six lactating women were recruited. The mean plasma protein was 8.17 g/dl, breastmilk protein was 3.36 g/dL, sIgA was 4.37 g/L and LF was 14.36 g/L. The total energy, carbohydrate, protein, fat and vitamin C and E daily intakes were 1828.61 kcal/day, 234.76, 71.15, 72.31 gram/day, 109.7 mg/day and 5.60 mg/day, respectively. Neither plasma and breastmilk total protein nor sIgA and LF concentrations were correlated with total energy, macronutrients or vitamin C and E daily intake. Conclusions: The plasma total protein, breastmilk total protein, sIgA and LF concentrations of lactating women in Jakarta were within the normal range. These biochemical profiles were shown to be not significantly correlated with daily calory, macronutrient or vitamin C or E intake. Keywords: breastmilk protein; secretory immunoglobulin A; lactoferrin; macronutrients; vitamins