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1.
Viruses ; 15(4)2023 03 30.
Article in English | MEDLINE | ID: mdl-37112867

ABSTRACT

Staphylococcus aureus causes intramammary infections (IMIs), which are refractory to antibiotic treatment and frequently result in chronic mastitis. IMIs are the leading cause of conventional antibiotic use in dairy farms. Phage therapy represents an alternative to antibiotics to help better manage mastitis in cows, reducing the global spread of resistance. A mouse mastitis model of S. aureus IMI was used to study the efficacy of a new cocktail of five lytic S. aureus-specific phages (StaphLyse™), administered either via the intramammary (IMAM) route or intravenously (IV). The StaphLyse™ phage cocktail was stable in milk for up to one day at 37 °C and up to one week at 4 °C. The phage cocktail was bactericidal in vitro against S. aureus in a dose-dependent manner. A single IMAM injection of this cocktail given 8 h after infection reduced the bacterial load in the mammary glands of lactating mice infected with S. aureus, and as expected, a two-dose regimen was more effective. Prophylactic use (4 h pre-challenge) of the phage cocktail was also effective, reducing S. aureus levels by 4 log10 CFU per gram of mammary gland. These results suggest that phage therapy may be a viable alternative to traditional antibiotics for the control of S. aureus IMIs.


Subject(s)
Bacteriophages , Mastitis, Bovine , Staphylococcal Infections , Female , Animals , Mice , Cattle , Staphylococcus aureus , Lactation , Mastitis, Bovine/drug therapy , Mastitis, Bovine/microbiology , Disease Models, Animal , Anti-Bacterial Agents/therapeutic use , Anti-Bacterial Agents/pharmacology , Milk/microbiology , Staphylococcal Infections/microbiology
2.
Porcine Health Manag ; 9(1): 7, 2023 Feb 14.
Article in English | MEDLINE | ID: mdl-36782292

ABSTRACT

BACKGROUND: The 2017 Veterinary Feed Directive eliminated the use of medically important antibiotics for growth promotion of food animals; thus, alternative growth promoters are highly desirable by food animal producers to enhance animal health and reduce pathogen colonization, including the human foodborne pathogen Salmonella. ß(1-3)(1-6)-D-glucan (ß-glucan) is a soluble fiber with prebiotic characteristics; it has been shown to modulate immune and intestinal functions that strengthen swine resistance to health challenges such as bacterial infections when supplemented in the diets of growing pigs. The current study evaluated the effects of a ß-glucan product on gut microbial community structure as well as Salmonella shedding and intestinal colonization. RESULTS: Five-week-old pigs were fed a ß-glucan amended diet at 500 g/ton (n = 13) or a non-amended control diet (n = 14) for three weeks, followed by inoculation of the 27 pigs with 1 × 109 colony forming units of Salmonella enterica serovar Typhimurium strain UK1. While remaining on the respective diets, fecal samples collected at 2, 4, 7, and 16 days post-inoculation (dpi) were similar for Salmonella shedding counts between the two diets. At 16 dpi, Salmonella counts were significantly lower in the cecal contents of the ß-glucan-fed pigs (P = 0.0339) and a trend towards a reduction was observed in the Peyer's patches region of the ileum (P = 0.0790) compared to the control pigs. Pigs fed ß-glucan for three weeks exhibited an increase in members of the Clostridia class in their fecal microbial communities, and after inoculation with Salmonella, several potentially beneficial microorganisms were enriched in the microbiota of ß-glucan-fed pigs (Lactobacillus, Ruminococcaceae, Prevotellaceae, Veillonellaceae, Bifidobacterium and Olsenella). CONCLUSION: Administration of ß-glucan altered the swine gut microbiome and reduced Salmonella colonization in the cecal contents.

3.
Vet Microbiol ; 278: 109648, 2023 Mar.
Article in English | MEDLINE | ID: mdl-36608625

ABSTRACT

Alternatives to antibiotics to improve animal performance, limit the negative impact of infectious disease, and/or reduce colonization with foodborne pathogens is a major focus of animal agricultural research. ß-glucans, a generally-recognized-as-safe (GRAS) product derived from various sources, are used in swine and can serve as both a prebiotic and/or stimulant of the immune system given the expression of ß-glucan receptors on immune cells. When supplied in the diet of nursery pigs, it is unclear how dietary additives, particularly those known to modulate immune status, impact immunogenicity and efficacy of mucosal-delivered vaccines. Salmonellosis is one of the most common bacterial foodborne infections in the United States, and consumption of contaminated pork is a major source of human infection. Reduction of foodborne Salmonella in pigs via vaccination is one strategy to reduce contamination risk and subsequently reduce human disease. We examined the ability of dietary ß-glucan to modulate fecal microbial diversity, and immunogenicity and efficacy of a mucosally-delivered, live-attenuated Salmonella vaccine during the nursery period. While dietaryß-glucan did modulate fecal alpha diversity, it did not alter the induction of peripheral Salmonella-specific IFN-γ secreting Tcells or Salmonella-specific IgA in oral fluids. In addition, vaccination reduced Salmonella enterica serovar Typhimurium fecal shedding and tissue colonization. Overall, addition of ß-glucan to the nursery diet of pigs impacted the microbiota but did not alter mucosal vaccine immunogenicity and efficacy.


Subject(s)
Salmonella Infections, Animal , Salmonella Vaccines , Swine Diseases , beta-Glucans , Swine , Humans , Animals , Immunogenicity, Vaccine , Salmonella Infections, Animal/microbiology , Diet , Salmonella typhimurium , Vaccines, Attenuated , Swine Diseases/microbiology
4.
Transl Anim Sci ; 5(2): txab072, 2021 Apr.
Article in English | MEDLINE | ID: mdl-34189415

ABSTRACT

Study objectives were to determine the effects of dietary live yeast (Saccharomyces cerevisiae strain CNCM I-4407; ActisafHR+; 0.25g/kg of feed; Phileo by Lesaffre, Milwaukee, WI) on growth performance and biomarkers of metabolism and inflammation in heat-stressed and nutrient-restricted pigs. Crossbred barrows (n = 96; 79 ± 1 kg body weight [BW]) were blocked by initial BW and randomly assigned to one of six dietary-environmental treatments: 1) thermoneutral (TN) and fed ad libitum the control diet (TNCon), 2) TN and fed ad libitum a yeast containing diet (TNYeast), 3) TN and pair-fed (PF) the control diet (PFCon), 4) TN and PF the yeast containing diet (PFYeast), 5) heat stress (HS) and fed ad libitum the control diet (HSCon), or 6) HS and fed ad libitum the yeast diet (HSYeast). Following 5 d of acclimation to individual pens, pigs were enrolled in two experimental periods (P). During P1 (7 d), pigs were housed in TN conditions (20 °C) and fed their respective dietary treatments ad libitum. During P2 (28 d), HSCon and HSYeast pigs were fed ad libitum and exposed to progressive cyclical HS (28-33 °C) while TN and PF pigs remained in TN conditions and were fed ad libitum or PF to their HSCon and HSYeast counterparts. Pigs exposed to HS had an overall increase in rectal temperature, skin temperature, and respiration rate compared to TN pigs (0.3 °C, 5.5 °C, and 23 breaths per minute, respectively; P < 0.01). During P2, average daily feed intake (ADFI) decreased in HS compared to TN pigs (30%; P < 0.01). Average daily gain and final BW decreased in HS relative to TN pigs (P < 0.01); however, no differences in feed efficiency (G:F) were observed between HS and TN treatments (P > 0.16). A tendency for decreased ADFI and increased G:F was observed in TNYeast relative to TNCon pigs (P < 0.10). Circulating insulin was similar between HS and TN pigs (P > 0.42). Triiodothyronine and thyroxine levels decreased in HS compared to TN treatments (~19% and 20%, respectively; P < 0.05). Plasma tumor necrosis factor-alpha (TNF-α) did not differ across treatments (P > 0.57) but tended to decrease in HSYeast relative to HSCon pigs (P = 0.09). In summary, dietary live yeast did not affect body temperature indices or growth performance and had minimal effects on biomarkers of metabolism; however, it tended to improve G:F under TN conditions and tended to reduce the proinflammatory mediator TNF-α during HS. Further research on the potential role of dietary live yeast in pigs during HS or nutrient restriction scenarios is warranted.

5.
Animals (Basel) ; 11(6)2021 May 31.
Article in English | MEDLINE | ID: mdl-34072877

ABSTRACT

This study aimed to investigate the effects of dietary LY or LY combined with ZnO supplementation on performance and gut health in nursery pigs. 192 Duroc × Landrace × Yorkshire piglets (weaned on d 32 of the age with 9.2 ± 1.7 kg BW) were allocated into four treatments with eight replicate pens, six piglets per pen. The treatments included a basal diet as control (CTR), an antibiotic plus ZnO diet (CTC-ZnO, basal diet + 75 mg/kg of chlortetracycline + ZnO (2000 mg/kg from d 1 to 14, 160 mg/kg from d 15 to 28)), a LY diet (LY, basal diet + 2 g/kg LY), and a LY plus ZnO diet (LY-ZnO, basal diet + 1 g/kg LY + ZnO). The results showed that pigs fed LY or LY-ZnO had increased (p < 0.05) average daily gain, serum IgA, IgG, superoxide dismutase, fecal butyric acid, and total volatile fatty acid concentrations, as well as decreased (p < 0.05) feed conversion ratio and diarrhea rate compared with CTR. In conclusion, pigs fed diets with LY or LY combined with ZnO had similar improvement to the use of antibiotics and ZnO in performance, antioxidant status, immunoglobulins, and gut health in nursery pigs.

6.
PLoS One ; 14(7): e0219557, 2019.
Article in English | MEDLINE | ID: mdl-31329605

ABSTRACT

One mechanism through which S. cerevisiae may improve the performance of pigs is by altering the composition of the gut microbiota, a response that may be enhanced by early postnatal supplementation of probiotics. To test this hypothesis, newborn piglets (16 piglets/group) were treated with either S. cerevisiae yeast (5 x 109 cfu/pig: Low) or (2.5 x 1010 cfu/piglet: High) or equivalent volume of sterile water (Control) by oral gavage every other day starting from day 1 of age until weaning (28±1 days of age). Piglet body weight was recorded on days 1, 3, 7, 10, 17, 24 and 28 and average daily gain (ADG) calculated for the total period. At weaning, piglets were euthanized to collect cecum content for microbial profiling by sequencing of the 16S rRNA gene. ADG was higher in both Low and High yeast groups than in Control group (P<0.05). Alpha diversity analyses indicated a more diverse microbiota in the Control group compared with Low yeast group; the High yeast being intermediate (P < 0.01). Similarly, Beta diversity analyses indicated differences among treatments (P = 0.03), mainly between Low yeast and Control groups (P = 0.02). The sparse Partial Least Squares Discriminant Analysis (sPLS-DA) indicated that Control group was discriminated by a higher abundance of Veillonella, Dorea, Oscillospira and Clostridium; Low yeast treated pigs by higher Blautia, Collinsella and Eubacterium; and High yeast treated pigs by higher Eubacterium, Anaerostipes, Parabacteroides, Mogibacterium and Phascolarctobacterium. Partial Least Squares (PLS) analysis showed that piglet ADG was positively correlated with genus Prevotella in High yeast group. Yeast supplementation significantly affected microbial diversity in cecal contents of suckling piglets associated with an improvement of short chain fatty acid producing bacteria in a dose-dependent manner. In conclusion, yeast treatment improved piglet performance and shaped the piglet cecum microbiota composition in a dose dependent way.


Subject(s)
Cecum/microbiology , Saccharomyces cerevisiae/physiology , Animals , Animals, Suckling , Biodiversity , Discriminant Analysis , Gastrointestinal Microbiome , Least-Squares Analysis , Phylogeny , Species Specificity , Swine/growth & development , Weight Gain
7.
J Anim Sci ; 97(4): 1806-1818, 2019 Apr 03.
Article in English | MEDLINE | ID: mdl-30796802

ABSTRACT

Two studies were conducted to investigate the effect of live yeast (LY) on the in vitro fermentation characteristics of wheat, barley, corn, soybean meal (SBM), canola meal, and distillers dried grains with solubles (DDGS). In Study 1, LY yeast was added directly to in vitro fermentations inoculated with feces from lactating sows, whereas as in study 2, feces collected from lactating sows fed LY as a daily supplement was used. Selected feedstuffs were digested and the residue added to separate replicated (n = 3) fermentation reactions. Study 1 was conducted in two blocks, whereas study 2 was conducted using feces collected after a period of 3 (Exp. 1) or 4 wk (Exp. 2) of LY supplementation. Accumulated gas produced over 72 h was modeled for each substrate and the kinetics parameters compared between LY and control groups. The molar ratio of the volatile fatty acids (VFAs) produced in vitro were also compared at 12 and 72 h of incubation. In study 1, in vitro addition of yeast increased (P < 0.001) the rate of gas production (Rmax). However, a yeast × substrate effect (P < 0.05) observed for total gas accumulated (A), time to half asymptote (B), and time required to reach maximum rate of fermentation (Tmax) suggested that yeast-mediated increases in extent and rate of fermentation varied by substrate. Greater total gas production was observed only for corn and SBM, associated with greater B and Tmax. Supplementation with LY appeared to increase A and Rmax although with variation between experiments and substrates. In Exp. 1, LY decreased (P < 0.05) B and Tmax. However, a yeast × substrate effect (P < 0.05) was observed for only A (for wheat, barley, corn, and corn DDGS) and Rmax (wheat, barley, corn, and wheat DDGS). In Exp. 2, LY increased (P < 0.0001) A and decreased B. However, an interaction (P < 0.05) with substrates was observed for Rmax (except SBM) and Tmax. With exception of the DDGS samples, LY supplementation increased (P < 0.05) VFA production at 12 and 72 h of incubation. Yeast increased (P < 0.05) the molar ratios of acetic acid and branch-chain fatty acids at 12 h of incubation; however, this response was more variable by substrate at 72 h. In conclusion, LY supplementation increased the rate and extent of in vitro fermentation of a variety of substrates prepared from common feedstuffs. Greater effects were observed when LY was fed to sows than added directly in vitro, suggesting effects on fermentation were not mediated directly.


Subject(s)
Animal Feed/analysis , Dietary Supplements , Gastrointestinal Microbiome/drug effects , Swine/physiology , Yeast, Dried/pharmacology , Animals , Diet/veterinary , Digestion/drug effects , Fatty Acids, Volatile/metabolism , Feces/microbiology , Female , Fermentation/drug effects , Hordeum , Lactation/drug effects , Probiotics , Glycine max , Swine/microbiology , Triticum , Zea mays
8.
Br J Nutr ; 118(11): 949-958, 2017 Dec.
Article in English | MEDLINE | ID: mdl-29166952

ABSTRACT

This study aimed to investigate the effects of dietary live yeast (LY) supplementation on growth, intestinal permeability and immunological parameters of piglets challenged with enterotoxigenic Escherichia coli K88 (ETEC). Piglets weaned at 21 d were allocated into three treatments with six pens and six piglets per pen, receiving the control diet (CON), diets supplemented with antibiotics plus zinc oxide (ANT-ZnO) and LY (Saccharomyces cerevisiae strain CNCM I-4407), respectively, for a period of 2 weeks. On day 8, thirty-six piglets were selected as control without ETEC (CON), CON-ETEC, ANT-ZnO-ETEC and LY-ETEC groups challenged with ETEC until day 10 for sample collections. Piglets fed ANT-ZnO diet had the highest average daily gain and average daily feed intake (P<0·05) during the 1st week, but ADG of piglets fed the ANT-ZnO diet was similar as piglets fed LY diet during the second week. Piglets with LY-ETEC or ANT-ZnO-ETEC had markedly lower diarrhoea score (P<0·05) than piglets with CON-ETEC during the 24 h after ETEC challenge. Relative to piglets with CON, the counts of E. coli, urinary ratio of lactulose to mannitol, plasma IL-6 concentration, mRNA abundances of innate immunity-related genes in ileum and mesenteric lymph node tissues were increased (P<0·05), whereas the villous height of jejunum and relative protein expression of ileum claudin-1 were decreased (P<0·05) in piglets with CON-ETEC; however, these parameters did not markedly change in piglets with LY-ETEC or ANT-ZnO-ETEC. In summary, dietary LY supplementation could alleviate the severity of diarrhoea in piglets with ETEC, which may be associated with the improved permeability, innate immunity and bacterial profile.


Subject(s)
Diarrhea/therapy , Dietary Supplements , Escherichia coli Infections/veterinary , Gastrointestinal Microbiome , Intestines/microbiology , Yeast, Dried/administration & dosage , Animals , Anti-Bacterial Agents/administration & dosage , Claudin-1/genetics , Claudin-1/metabolism , Colony Count, Microbial , Diarrhea/microbiology , Diarrhea/veterinary , Diet/veterinary , Enterotoxigenic Escherichia coli , Escherichia coli Infections/therapy , Gene Expression Regulation , Interleukin-1beta/genetics , Interleukin-1beta/metabolism , Interleukin-6/genetics , Interleukin-6/metabolism , Male , Myeloid Differentiation Factor 88/genetics , Myeloid Differentiation Factor 88/metabolism , NF-kappa B/genetics , NF-kappa B/metabolism , Permeability , Swine , Weaning , Zinc Oxide/administration & dosage
9.
Vet Immunol Immunopathol ; 144(3-4): 448-54, 2011 Dec 15.
Article in English | MEDLINE | ID: mdl-21955445

ABSTRACT

Interleukin-17 (IL-17) producing cells, referred to as Th17, have recently emerged as a third subset of the T helper (Th) cell family. Studies in mice have demonstrated that Th17 cells and their associated cytokines are involved in several autoimmune diseases and host defense against infection. Murine Th17 cells differentiate from naïve CD4(+) T-cells in the presence of TGFß and IL-6, however, there are contradicting reports as to the role of TGFß in the differentiation of human Th17 cells and very little is known about these cells in other animals. We report here the presence of IL-17 secreting lymphocytes in the lung and peripheral blood of pigs. The cDNA of porcine IL-17 gene was cloned and sequenced from activated lung lymphocytes and PBMC from piglets. A 17kDa recombinant protein was expressed and purified both under denaturing and native conditions from E. coli BL21 cells. Furthermore, we demonstrate that TGFß in the presence of IL-6 and/or IL-1ß induces in vitro differentiation of Th17 cells from naïve porcine CD4(+) thymocytes.


Subject(s)
Interleukin-17/metabolism , Leukocytes, Mononuclear/cytology , Lung/cytology , Swine/immunology , Th17 Cells/metabolism , Thymus Gland/cytology , Amino Acid Sequence , Animals , Base Sequence , Cloning, Molecular , Interleukin-17/genetics , Interleukin-17/physiology , Interleukin-1beta/physiology , Interleukin-6/physiology , Leukocytes, Mononuclear/immunology , Leukocytes, Mononuclear/physiology , Lung/immunology , Lymphocyte Activation , Molecular Sequence Data , Real-Time Polymerase Chain Reaction/veterinary , Th17 Cells/immunology , Th17 Cells/physiology , Thymus Gland/immunology , Transforming Growth Factor beta/physiology
10.
Vaccine ; 29(38): 6540-8, 2011 Sep 02.
Article in English | MEDLINE | ID: mdl-21771625

ABSTRACT

We investigated the efficacy of a novel microparticle (MP) based vaccine formulation consisting of pertussis toxoid (PTd), polyphosphazene (PCEP), CpG ODN 10101 and synthetic cationic innate defense regulator peptide 1002 (IDR) against Bordetella pertussis in mice. We studied whether encapsulation of these IDR-CpG ODN complexes into polyphosphazene-based microparticles further enhanced their immunomodulatory activity compared to soluble formulations containing PCEP (SOL), or without PCEP (AQ). In vitro stimulation of murine macrophages showed MP induced significantly higher levels of pro-inflammatory cytokines. When assessed in a B. pertussis infection challenge model, a single immunization with MP formulation led to significantly lower bacterial loads compared to other formulations and non-vaccinated animals. ELISPOT of splenocytes showed that MP group mice had significantly higher number of antigen-specific IL-17 secreting cells. The cytokine profile in lung homogenates of MP group mice after challenge showed significantly higher amounts of MCP-1, TNF-α, IFN-γ, IL-12 and IL-17 and significantly lowered IL-10 levels suggesting a strong Th1 shift. Protection was observed against challenge infection with B. pertussis. On the other hand protective immune responses elicited in Quadracel(®) immunized mice were Th2 skewed. Hence, we conclude that formulation of PTd, PCEP, CpG ODN and IDR into MP generates a protective immune response in mice against pertussis emphasizing the potential of MP as a delivery vehicle for the potential development of single-shot vaccines.


Subject(s)
Antimicrobial Cationic Peptides/administration & dosage , Drug Carriers/administration & dosage , Oligodeoxyribonucleotides/administration & dosage , Pertussis Vaccine/immunology , Phenylpropionates/administration & dosage , Polymers/administration & dosage , Toxoids/administration & dosage , Whooping Cough/prevention & control , Adjuvants, Immunologic/administration & dosage , Animals , Bacterial Load , Cytokines/metabolism , Disease Models, Animal , Female , Lung/microbiology , Lymphocytes/immunology , Macrophages/immunology , Mice , Mice, Inbred BALB C , Microspheres , Pertussis Vaccine/administration & dosage , Rodent Diseases/prevention & control , Spleen/immunology
11.
Immunotherapy ; 2(1): 25-35, 2010 Jan.
Article in English | MEDLINE | ID: mdl-20635889

ABSTRACT

Neonatal bacille Calmette-Guerin (BCG) vaccination is widely employed to protect against tuberculosis. Predominant Th1 but not mixed Th1/Th2 responses are thought to be protective. If so, effective vaccination must cause Th1 imprints. The immune system of infants differs from that of adults and such differences could critically affect neonatal vaccination. We demonstrate that BCG infection of infant and adult mice produces similar responses. Infection with low and high numbers of BCG, respectively, leads to sustained Th1 and mixed Th1/Th2 responses. Low-dose but not high-dose infection also results in Th1 imprints, guaranteeing a Th1 response upon high-dose challenge, and resulting in optimal bacterial clearance. Our observations on low-dose Th1 imprinting are intriguing in the context of the well-known madras trial. In this trial, the highest dose of BCG, which had insignificant side effects, was administered to over 250,000 human subjects. This high-dose vaccination resulted in insignificant protection against tuberculosis.


Subject(s)
BCG Vaccine/immunology , Mycobacterium bovis/immunology , Th1 Cells/immunology , Tuberculosis/immunology , Animals , Animals, Newborn , BCG Vaccine/administration & dosage , Dose-Response Relationship, Drug , Enzyme-Linked Immunosorbent Assay , Female , Immunization , Interferon-gamma/metabolism , Interleukin-4/metabolism , Mice , Mice, Inbred BALB C , Spleen/immunology , Spleen/metabolism , Th1 Cells/metabolism , Time Factors , Tuberculosis/prevention & control , Tuberculosis/veterinary
12.
Vet Immunol Immunopathol ; 130(3-4): 251-5, 2009 Aug 15.
Article in English | MEDLINE | ID: mdl-19178951

ABSTRACT

Changes in the proportion of peripheral blood T cell subsets after subcutaneous inoculation of cattle with Mycobacterium bovis Bacille Calmette-Guerin (BCG) were studied. Calves were injected with approximately 8 x 10(6) BCG bacillus and blood samples collected at weekly intervals for flow-cytometric analyses to determine the proportion of CD4+, CD8+ and gammadelta T cells. In addition, whole blood samples were stimulated in vitro with M. bovis purified protein derivative (PPD) and the secreted IFN-gamma quantified by ELISA. Results showed cellular and cytokine changes which could be categorized into three phases. The first phase occurred within the first 2 weeks after vaccination involving an increase in proportion of WC1+ gammadelta T cells and a concomitant increase in the secretion of IFN-gamma. These two responses peaked at 2 weeks and waned thereafter. The second phase involved an increase in the CD4/CD8 ratio as a result of an increase in the proportion of CD4+ T cells between 4 and 6 weeks. The third phase involved a decrease in the CD4/CD8 ratio due to an increase in the proportion of CD8+ T cells between 8 and 10 weeks. Surprisingly, the IFN-gamma response was associated with changes in the gammadelta rather than the CD4+ or CD8+ T cells, suggesting that this cytokine was secreted by gammadelta-T cells. These results are consistent with the reported ability of gammadelta T cells to act rapidly and bridging the innate and classically adaptive immune responses.


Subject(s)
BCG Vaccine/administration & dosage , Cattle/immunology , Receptors, Antigen, T-Cell, gamma-delta/metabolism , T-Lymphocyte Subsets/immunology , Animals , BCG Vaccine/immunology , CD4-CD8 Ratio , Immunity, Innate , In Vitro Techniques , Interferon-gamma/biosynthesis , Time Factors , Tuberculin/administration & dosage
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