Effect of freeze-dried powder of lactic acid bacteria treatment of pediatric bacterial peritonitis on immune function and nutritional status

Abstract

Objectives

This study was to investigate the effects of freeze-dried powder of lactic acid bacteria treatment of pediatric bacterial peritonitis on immune function and nutritional status.

Methods

100 cases of bacterial peritonitis children from September 2020 to September 2021 were included and divided into a control group and an observation group using a randomized numerical table, with each group containing 50 cases. The control group was given routine treatment such as antimicrobial drugs, and the observation group was given adjuvant treatment of freeze-dried powder of lactic acid bacteria. The clinical efficacy, immune function, nutritional status, inflammatory response, intestinal dysbiosis, and endotoxin level were compared between the two groups.

Results

The total clinical effective rate of the observation group was higher than that of the control group. The ratio of CD3⁺, CD4⁺, and CD4⁺/CD8⁺ and the levels of ALB, TP, and Hb after treatment of the observation group were higher than those of the control group. CRP, PCT, and MCP-1 of the observation group were lower than those of the control group after treatment. Intestinal flora dysbiosis in the observation group was attenuated, and endotoxin level was reduced compared to the control group. No drug-related adverse reactions were observed in both groups during treatment.

Conclusions

Freeze-dried powder of lactic acid bacteria can improve immune function and nutritional status, as well as reduce inflammatory response and intestinal flora dysbiosis in pediatric bacterial peritonitis.

Introduction

Pediatric bacterial peritonitis is caused by surgical infection, gastrointestinal perforation, and trauma, with clinical manifestations such as fever, nausea, vomiting, and abdominal pain, and in severe cases, toxic shock and other symptoms may occur [1]. It is currently believed that the development of pediatric bacterial peritonitis is affected by the type and nature of the cause, the location of the primary lesion, the type and number of bacteria, and the host's defense ability, so the immune function is essential for the recovery of the organism [2]. In medical environments, antibacterial drugs are frequently used to treat this condition. Although they can ease patient symptoms, the extended course of the disease and other constraints require the use of additional medications [3]. Live bacteria that are physiologically active are vital in treating various abdominal conditions, including intestinal tract dysbiosis [4]. Powder of Lactic Acid Bacteria is a probiotic preparation with a variety of probiotic components, such as Lactobacillus plantarum JYLP-375 and bifidobacterium lactis Bi-07, which can directly activate the immune response of the intestinal mucosa and then have an effect on the immune function of the mucosa in other parts of the body [5]. Therefore, this study was to investigate the effect of Powder of Lactic Acid Bacteria adjuvant treatment of pediatric bacterial peritonitis on immune function and nutritional status and to provide a theoretical basis for the treatment of this disease in the clinic.

Materials and methods

Clinical data

100 cases of children with bacterial peritonitis from September 2020 to September 2021 were studied, and they were allocated into a control group and an observation group using a randomized numerical table, with each group comprising 50 cases.

Diagnostic criteria

In these children, paracentesis was the standard method of collecting ascites [6]. An abdominal puncture yielded 10 mL of ascites, which was then placed in a blood culture bottle. Ascites samples were placed in Blood Agar and MacConckey Agar, and preliminary results were obtained after 48 h. The diagnostic criteria for bacterial peritonitis [7] were: (1) positive gram staining of ascites bacterial cultures within 48 h of admission (no intra-abdominal source of infection); (2) absolute count of polymorphonuclear leukocytes (PMN) ≥ 250 cells/mm³ in the ascites; and (3) abdominal pain, fever, vomiting, or anorexia.

Inclusion criteria

① Compliance with the diagnostic criteria for pediatric bacterial peritonitis; ② Informed consent obtained from the families of the children; ③ Children aged 3 ~ 14 years.

Exclusion criteria

① Children with severe cardiac, hepatic, and renal function abnormalities; ② Children with combined immune function disorders; ③ Children with combined other infectious diseases; ④ Children with combined congenital diseases; ⑤ Children who were allergic to the drugs used in this study; ⑥ Children with combined chronic inflammatory diseases; ⑦ Children with combined other intestinal diseases.

Methods

The control group was given antibacterial drugs and other conventional treatments, and the observation group was additionally given Powder of Lactic Acid Bacteria adjuvant treatment using MONPELLERIN8 freeze-dried powder of lactic acid bacteria (202,100,005, Yantai Maijixide Bioengineering Co., Ltd., specifications: ≥ 1.5 × 10¹¹/bag, 3 g/bag, live bacteria per bag > 150 billion CF [5 × 10¹⁰ CFU/g]). The freeze-dried powder was dissolved in warm water at 37 ℃ and administrated orally (3 g/times, 1 time/day, a total of 7 day treatment).

Observation parameters

Tests for rapid response indicators, such as inflammatory response parameters and endotoxin, were synchronized before drug administration and 24–48 h post-treatment. For indicators such as immune function parameters, intestinal dysbiosis grading, and nutritional status, where there might be a time to onset of action, tests were conducted before the drug was given and again 7 days after the treatment concluded. To determine clinical efficacy, symptoms must be tested daily and recorded, followed by a summary evaluation after a week. (1) Clinical efficacy was evaluated as follows [8]: obviously effective: clinical symptoms such as abdominal pain, pressure, muscle tension, and rebound pain disappeared, and laboratory parameters such as inflammatory parameters returned to normal; effective: clinical symptoms and laboratory parameters were significantly improved; ineffective: no improvement or aggravation of clinical symptoms and laboratory parameters was detected. (2) Immune function parameters: T-lymphocyte subpopulation ratio was detected by flow cytometry, and the reagents were provided by BD Company (USA). (3) Nutritional status: the 7600 automatic biochemical analyzer (Hitachi, Japan) and the xs800i and xe2100D blood cell analyzers (Sysmex, Japan) were utilized. Serum ALB level was determined by the bromocresol green method, serum TP level was determined by the Biuret method, and Hb in the whole blood was determined by the sodium dodecyl sulfate method. (4) Inflammatory response parameters: using the AD-VIA2400 Siemens automatic biochemical analyzer (SIMENS, Germany), serum monocyte chemoattractant protein-1 (MCP-1) level was detected by ELISA kit (R&D Company, USA), serum C-reactive protein (CRP) level was detected by immunoturbidimetric method using the kit (Fujian Sandi Biotechnology Co., Ltd., Fujian, China), and serum procalcitonin (PCT) level was measured by chemiluminescence assay using the kit (Tarcine BioMed Inc., Beijing, China). (5) Intestinal flora dysbiosis [9]: fresh stool that was naturally discharged and uncontaminated was collected and used to create fecal smears for gram staining. Total bacterial count, morphological characteristics, and coccus/bacillus ratio were detected microscopically by the same experienced examiner in half an hour to classify intestinal flora dysbiosis into mild, moderate, and severe (I, II, and III grades, respectively). The grading standard is shown in Table 1. (6) Endotoxin level: serum endotoxin was detected by immunoturbidimetric method using the kit (Beijing Gold Mountainriver Tech Development Co., Ltd., Beijing, China).

Table 1.

Degree of intestinal dysbiosis

Degree of intestinal dysbiosis	total number of bacteria	G +/G bacillus	G + cocci	Cocci/Bacilli ratio	yeast-like fungi and clostridium butyricum
Normal	500 ~ 5000/oil len field	G + bacilli were more than G bacilli	Few G + cocci	About 25/75	No
I (mild)	Low normal value or slightly reduced, 101 ~ 500/oil len field	Both were in normal low value or mild reduction	High normal values or mildly–moderately increased	Mildly altered bacterial ratios, with an increase in cocci and a decrease in bacilli	Slightly increased
II (moderate)	Significantly reduced, 11 ~ 100/oil len field	Both significantly reduced	Markedly increased	Significantly altered bacterial ratios with inverted cocci/bacteria ratios	Markedly increased
III (severe)	Markedly reduced, < 10/oil len field	Most of the original flora is suppressed. Only one bacterium or fungus is absolutely dominant. A certain minority of the original bacteria becomes absolutely dominant

Statistical analysis

The data obtained from the study were processed by SPSS 22.0 software. Enumeration data were expressed as percentages and comparatively analyzed by the χ² test. Measurement data were expressed as $\overline{x}\pm \text{s}$ after the normal test and assessed by t test. Rank data were compared by rank-sum test. Effect sizes for both within and between groups were calculated using paired samples t tests and independent samples t tests, respectively, with Cohen's d values chosen for reporting. The differences were statistically significant at P < 0.05.

Results

Clinical data

There was no significant difference in the comparison of clinical data between the two groups (P > 0.05, Table 2).

Table 2.

Comparison of clinical data between the two groups

Data	Observation group	Control group	Cohen’s d	P value
Gender (cases)				0.84
Male	29	27
Female	21	23
Age (years)	7.14 ± 1.58	7.20 ± 1.71	− 0.04	0.86
Disease course (days)	2.96 ± 0.59	3.01 ± 0.54	− 0.09	0.66
Primary diseases (cases)				0.80
Surgical infectious diseases	21	18
Digestive tract perforation	19	20
Trauma	10	12

Surgical infectious diseases includes acute appendicitis with perforation, peritoneal abscess, and necrotizing enterocolitis; Digestive tract perforation includes perforation secondary to intestinal obstruction, inflammatory bowel disease perforation, and peptic ulcer perforation; and trauma includes rupture of bowel tubes caused by external forces, penetrating trauma and medically induced injury

Clinical efficacy

The total clinical efficacy rate of the observation group was higher than that of the control group (P < 0.05, Table 3).

Table 3.

Comparison of clinical efficacy between the two groups (cases, %)

Groups	n	Obviously effective	Effective	Ineffective	Total effective rate
Observation group	50	21	27	2	96
Control group	50	16	25	9	82
χ2 value					5.005
P value					0.025

Immune function parameters

The immune function metrics of the two groups were comparable before treatment, with no significant differences (P > 0.05). The observation group showed higher ratios of CD3⁺ (Cohen’s d = 0.85, P < 0.001), CD4⁺ (Cohen’s d = 0.95, P < 0.001), and CD4⁺/CD8⁺ (Cohen’s d = 1.04, P < 0.001) compared to the control group after treatment (Fig. 1).

Fig. 1.

Comparison of immune function parameters between the two groups. (#P < 0.05 compared with before treatment; *P < 0.05 compared with control group after treatment)

Nutritional status

The nutritional status metrics of the two groups did not differ significantly before treatment (P > 0.05). ALB (Cohen’s d = 0.85, P < 0.001), TP (Cohen’s d = 0.78, P = 0.00), and Hb (Cohen’s d = 0.82, P < 0.001) of the observation group were higher than those of the control group after treatment (Fig. 2).

Fig. 2.

Comparison of the nutritional status of the two groups. (#P < 0.05 compared with before treatment; *P < 0.05 compared with control group after treatment)

Inflammatory response parameters

Inflammatory response parameters before and after treatment did not differ significantly between the two groups (P > 0.05). CRP (Cohen’s d = − 0.81, P = 0.00), PCT (Cohen’s d = − 1.70, P < 0.001), and MCP-1 (Cohen's d = − 1.11, P < 0.001) were lower in the observation group than in the control group after treatment (Fig. 3).

Fig. 3.

Comparison of inflammatory response parameters between the two groups. (#P < 0.05 compared with before treatment; *P < 0.05 compared with control group after treatment)

Dysbiosis

In the observation group, the severity of intestinal dysbiosis (Cohen's d = − 2, P < 0.001) was lower than in the control group (Table 4).

Table 4.

Comparison of the degree of dysbiosis between the two groups before and after treatment (n = 50)

Groups		I	II	III	Z value	P value
Observation group	Before treatment	9	28	13	10.678	0.001
	After treatment	21	26	3
Control group	Before treatment	10	26	14	4.871	0.027
	After treatment	16	29	5

Endotoxin levels

Endotoxin levels before treatment were not significantly different between the two groups (P > 0.05). Endotoxin levels in the observation group were lower than those in the control group after treatment (P < 0.05, Fig. 4).

Fig. 4.

Comparison of endotoxin levels between the two groups before and after treatment. (#P < 0.05 compared with before treatment; *P < 0.05 compared with control group after treatment)

Adverse reactions

No drug-related adverse reactions were observed in both groups during treatment.

Discussion

Pediatric peritonitis pathologically originates from the peritoneal wall layer or visceral layer, often triggered or damaged by acute inflammatory responses, primarily due to bacterial infections, chemical stimuli, or physical injury. In children, peritonitis is primarily indicated by symptoms, such as peritonitis triad-abdominal pressure pain, muscle tension, and rebound pain, along with nausea, vomiting, fever, and elevated white blood cells. In extreme cases, it can result in decreased blood pressure and widespread toxic effects [10]. Antimicrobial drugs are commonly used to treat this illness, as they are effective in eradicating pathogens and reducing the impact of harmful bacteria on the intestines and other abdominal areas [11]. However, it has been reported that the administration of probiotics in combination with antimicrobial drugs can promote the recovery of intestinal function in patients [12]. This study found that the total clinical effective rate of the observation group was higher than that of the control group, which further confirmed that the adjuvant treatment with Powder of Lactic Acid Bacteria could effectively improve the outcome of patients. This study also found that the severity of intestinal flora dysbiosis in the observation group was lower than that in the control group, indicating that the adjuvant treatment of pediatric bacterial peritonitis with Powder of Lactic Acid Bacteria can alleviate intestinal flora dysbiosis, which is mainly due to the active ingredients of Powder of Lactic Acid Bacteria in the human intestinal tract. By invigorating the lymphatic system, Lactic Acid Bacteria powder helps create mucosal barriers, enhances the body's ability to resist pathogens, and slows the growth and spread of harmful bacteria and different metabolites, thereby directly eliminating bacteria and potentially easing intestinal flora ailments [13].

It is currently believed that children are immunocompromised and prone to prolonged bacterial peritonitis. The intestinal tract is one of the immune organs of the human body, and intestinal flora is an important part of the intestinal immune system [14]. This study determined that Powder of Lactic Acid Bacteria treatment can effectively improve intestinal flora disorders, suggesting that it may improve the immune function of patients to a certain extent. The results showed that after treatment, the observation group had higher ratios of CD3 +, CD4 +, and CD4 +/CD8 + compared to the control group, indicating that Lactic Acid Bacteria Powder can enhance patients’ immune function. This is due to Powder of Lactic Acid Bacteria being beneficial microorganisms for human health, influencing the equilibrium of gut flora. This regulation affects both the mucosal and gastrointestinal immune systems, thereby impacting the overall immune response [15, 16]. Powder of Lactic Acid Bacteria contains various probiotic components, such as Lactobacillus plantarum JYLP-375, Bifidobacterium lactis Bi-07, etc., which can directly activate the immune response of the intestinal mucosa.

During the early phase of peritonitis, there is a rise, subsequent weakening, and eventual onset of intestinal paralysis, characterized by heightened intestinal secretion, reduced absorption, and substantial gas and fluid accumulation in the intestinal lumen. The intestinal wall, peritoneum, and mesentery become edematous, with a large amount of inflammatory exudate entering the abdominal cavity, resulting in water, electrolyte, and protein loss, as well as malnutrition [17]. Under normal conditions, intestinal flora can block the invasion of pathogenic microorganisms, but under pathological conditions, they disrupt the gastrointestinal system and influence iron absorption [18]. This study determined that ALB, TP, and Hb levels of the observation group were higher than those of the control group after treatment, indicating that Powder of Lactic Acid Bacteria can improve nutritional status of patients. The reason for this is that Powder of Lactic Acid Bacteria offers anti-infection, immune-enhancing, and anti-aging benefits, and can secrete digestive enzymes to convert intolerable glucose, fats, and proteins into soluble, easily absorbed substances, thereby improving nutritional status [18].

The intestinal tract can continuously release endotoxin, and the normal mechanical barrier, biological barrier, and immune barrier of the intestinal mucosa prevent endotoxin absorption. Pathogenic bacteria invading the pediatric intestinal tract can adhere to the intestinal epithelium and multiply extracellularly to produce enterotoxins, causing diarrhea. Endotoxin is the strongest stimulus to induce TNF-a production, due to the stimulation of bacteria, endotoxin, inflammation, etc., activating the body macrophages and T cells, etc. and releasing a variety of cytokines, such as inflammatory mediators [19]. The peritoneum exhibits an instant inflammatory reaction, including congestion, edema, and exudate, once the abdominal cavity becomes infected. Fibrin found in the exudate supports the adhesion of intestinal collaterals and the greater momentum to the inflamed peritoneal region, thus limiting the extension of the inflammation [20]. However, failure to remove the infected lesion may induce diffuse peritonitis due to excessive virulence and bacteria. This study revealed that CRP, PCT, MCP-1, and endotoxin in the observation group were lower than those in the control group after treatment, indicating that the adjuvant treatment with Powder of Lactic Acid Bacteria could reduce inflammatory response, which is mainly related to its ability to reduce the multiplication rate of harmful bacteria and invasion rate, as well as to reduce the disturbance of the intestinal flora and to improve the immune function of the organism [21]. In addition, no drug-related adverse reactions occurred during treatment in both groups, indicating that the safety of both conventional treatment and Powder of Lactic Acid Bacteria treatment is high.

In conclusion, the adjuvant treatment of pediatric bacterial peritonitis with Powder of Lactic Acid Bacteria can improve immune function and nutritional status, as well as reduce inflammatory response and intestinal flora dysbiosis.

Author contributions

Conceptualization, Yang Xue; methodology, Yang Xue and YuTing Zhang; formal analysis, Yang Xue and YuTing Zhang; investigation, Yang Xue and YuTing Zhang; data curation, Yang Xue and YuTing Zhang; writing—original draft preparation, Yang Xue; writing—review and editing, YuTing Zhang; project administration, YuTing Zhang. All authors have read and agreed to the published version of the manuscript.

Funding

Not applicable.

Availability of data and materials

The data that support the findings of this study are available from the corresponding author, upon reasonable request.

Declarations

Ethics approval and consent to participate

The present study was approved by the Ethics Committee of The First Hospital of China Medical University (No.201909LN-5) and written informed consent was provided by all patients prior to the study start. All procedures were performed in accordance with the ethical standards of the Institutional Review Board and The Declaration of Helsinki, and its later amendments or comparable ethical standards.

Competing interests

The authors declare no competing interests.