Bacitracin agar vs. oleandomycin disk supplemented chocolate agar for the recovery of Haemophilus influenzae in diagnostic samples: A prospective comparison.

Haemophilus influenzae is an important pathogen able to cause various forms of respiratory and invasive disease. To provide high sensitivity for detection, culture media must inhibit growth of residential flora from the respiratory tract. This study aimed to identify and compare the diagnostic and economic advantages of using bacitracin containing selective agar (SEL) or oleandomycin disk supplemented chocolate agar (CHOC). Growth and semi-quantitative abundance of H. influenzae and growth suppression of residential flora was prospectively assessed in a 28-week period. H. influenzae was identified in 164 (5 %) of all included samples: CHOC and SEL, CHOC only, and SEL only were positive in 95, 24, and 45 cases. Diagnostic superiority of SEL was primarily attributable to the results of throat swabs. However, on average, € 200 had to be spent for the detection of each additional isolate that was recovered only because of additional incubation on SEL.

Comparative effects of benzoic acid and sodium benzoate in diets for nursery pigs on growth performance and acidification of digesta and urine.

The objective of this study was to evaluate the comparative effects of benzoic acid and sodium benzoate in feeds on digesta pH, urinary pH, and growth performance for nursery pigs. A total of 432 pigs (6.9 ±â€…0.9 kg BW) were assigned to eight treatments (6 pigs per pen, replication = 9) in a randomized complete block design with initial body weight (BW) as a block and fed for 41 d in three phases (7/17/17 d, respectively). Treatments were 1) a basal diet (NC), 2) NC + 0.25% bacitracin methylene disalicylate (antibiotic; bacitracin: 250 g/t feed; PC), 3) NC + 0.25% benzoic acid, 4) NC + 0.35% benzoic acid, 5) NC + 0.50% benzoic acid, 6) NC + 0.30% sodium benzoate, 7) NC + 0.40% sodium benzoate, and 8) NC + 0.60% sodium benzoate. Growth performance and fecal scores were measured for each phase. One gilt representing the median BW of each pen was euthanized to collect digesta from the stomach, proximal jejunum, distal jejunum, and cecum, and urine. The PC tended to improve average daily gain (ADG) in phase 1 (P = 0.052) and phase 2 (P = 0.093) as well as average daily feed intake (ADFI) in phase 2 (P = 0.052). Overall, increasing supplemental benzoic acid tended to have a quadratic effect on ADG (P = 0.094), but no difference in ADFI was observed. Increasing supplemental sodium benzoate showed a quadratic effect (P < 0.05) on ADG and linearly increased (P < 0.05) ADFI. Urinary pH linearly decreased (P < 0.05) with increasing supplemental benzoic acid, but was not affected by supplemental sodium benzoate. Increasing supplemental benzoic acid or sodium benzoate linearly increased (P < 0.05) benzoic acid content in digesta of the stomach. Increasing supplemental benzoic acid or sodium benzoate also linearly increased (P < 0.05) urinary hippuric acid. However, the PC did not decrease urinary pH or increase urinary benzoic acid and hippuric acid. With slope-ratio assay using ADG and urinary hippuric acid as dependent variables and benzoic acid intake as an independent variable, the relative bioavailability of benzoic acid compared to sodium benzoate was not different. In conclusion, supplementation of benzoic acid and sodium benzoate could improve the growth performance of nursery pigs. The relative bioavailability of sodium benzoate to benzoic acid of nursery pigs did not differ based on BW gain and urinary hippuric acid.

Newly weaned pigs are exposed to various challenges during the postweaning period, resulting in retarded growth performance. Dietary antibiotics have been used to reduce the negative impacts of weaning stress. However, use of antibiotics in feeds has been phased out in response to concerns associated with microbial resistance. In this study, dietary benzoic acid was supplemented to promote growth performance and increase urinary hippuric acid of nursery pigs. The sodium benzoate may show similar effects with benzoic acid on growth performance and urinary hippuric acid, as sodium benzoate can be highly converted to benzoic acid via the action of gastric acid in stomach. Thus, this study aimed to investigate the effects of increasing benzoic acid and sodium benzoate supplementation on growth performance and acidification of digesta and urine, and to investigate the comparative effects of benzoic acid and sodium benzoate supplemented in diets for nursery pigs. Dietary benzoic acid and sodium benzoate improved body weight gain and increased urinary hippuric acid of nursery pigs. Both sodium benzoate and benzoic acid had similar effects when fed to nursery pigs for their body weight gain and metabolism. Benzoic acid, however, had a stronger effect acidifying urine compared with sodium benzoate.

Efficacy of disinfection procedures performed prior to regenerative endodontic therapy: An integrative review.

The aim of this integrative review was to identify whether the disinfection procedures performed prior to regenerative endodontic treatment were effective on biofilm removal from the root canals. The research was based on PubMed, Latin American and Caribbean Health Sciences Literature (Lilacs) and Scientific Electronic Library Online (SciELO) databases. Four articles were selected; one of the studies was in vivo and the others ex vivo. Different disinfection procedures were studied, characterised mainly by the use of intracanal medication, highlighting the double antibiotic paste, triple antibiotic paste and calcium hydroxide paste. Disinfection ability was evaluated against Enterococcus faecalis and multispecies biofilms by using the fluorescence technique and colony forming unit counting, for 7 to 21 days. Double antibiotic paste and triple antibiotic paste demonstrated excellent antibiofilm activity, unlike CH paste that showed limited disinfection, even when associated with different antimicrobial agents. Triple antibiotic paste was the most effective medication against biofilm.

The replacement of bacitracin methylene disalicylate with Bacillus subtilis PB6 in the diet of male Cherry Valley Ducks reduces the feed conversion ratio by improving intestinal health and modulating gut microbiota.

In this study, we compared the impacts of Bacillus subtilis PB6 (BS) and bacitracin methylene disalicylate (BMD) on the growth performance, intestinal morphology, expression of tight connection protein, and cecal microbiota community of male ducks through a 42-d trial. Three-hundred and sixty male Cherry Valley meat-type ducklings (1-day-old) were distributed into 3 groups of 6 replicates: CON group (control, basal diet), BMD group (basal diet + 45 mg/kg BMD, active ingredient dose in the feed), and BS group (basal diet + 2 × 107 CFU/kg BS in the feed). Results showed that supplementing the diet with BS reduced the average daily feed intake (ADFI) during d 15 to 42 and d 1 to 42 compared with the CON group (P = 0.032). It also reduced feed conversion ratio (FCR) during d 15 to 42 and d 1 to 42 (P < 0.05) relative to the other groups. The ileal villus height (VH) and villus height /crypt depth ratio (V/C) were increased (P < 0.05) in both the BS and BMD groups, and the jejunal VH and V/C ratio were increased in the BS group (P < 0.05). Relative to the CON, BS supplementation was associated with numerical augmentation of goblet cells in the jejunal mucosa and upregulation of jejunal zonula occludens (ZO-1) and ileal mucin2 (P < 0.05) mRNA levels. Analysis showed a negative correlation between FCR (d 0-42) and VH, V/C, and the number of goblet cells in the jejunum (P < 0.05). Additionally, BMD or BS supplementation altered the alpha diversity of colonic microbiota (P < 0.05). Correlation analysis revealed that Butyricimonas, Enterobacteriaceae, Clostridiaceae, and Tannerellaceae were positively associated with the acetic acid and butyrate concentrations (P < 0.05). Taken together, the supplementation of BS in the diet of male ducks was conducive to reducing FCR by meliorating intestinal morphology, upregulating ZO-1 and mucin2 mRNA levels, regulating the abundance of microbiota, and metabolites, and having a greater effect than BMD supplementation.

A mixture of organic acids and thymol protects primary chicken intestinal epithelial cells from Clostridium perfringens infection in vitro.

Necrotic enteritis causes economic losses estimated to be up to 6 billion US dollars per year. Clinical and subclinical infections in poultry are also both correlated with decreased growth and feed efficiency. Moreover, in a context of increased antibiotic resistance, feed additives with enhanced antimicrobial properties are a useful and increasingly needed strategy. In this study, the protective effects of a blend of thymol and organic acids against the effects of Clostridium perfringens type A (CP) on chicken intestinal epithelial cells were investigated and compared to bacitracin, a widely used antibiotic in poultry production. Primary chicken intestinal epithelial cells were challenged with CP for a total time of 3 h to assess the beneficial effect of 2 doses of citric acid, dodecanoic acid, and thymol-containing blend, and compare them with bacitracin. During the challenge, different parameters were recorded, such as transepithelial electrical resistance, cell viability, mRNA expression, and reactive oxygen species production. CP induced inflammation with cytokine production and loss of epithelial barrier integrity. It was also able to induce reactive oxygen species production and increase the caspase expression leading to cellular death. The high dose of the blend acted similarly to bacitracin, preventing the disruptive effects of CP and inducing also an increase in zonula occludens-1 mRNA expression. The low dose only partially prevented the disruptive effects of CP but successfully reduced the associated inflammation. This study shows that the usage of thymol combined with 2 organic acids can protect primary chicken intestinal epithelial cells from CP-induced damages creating a valid candidate to substitute or adjuvate the antibiotic treatment against necrotic enteritis.

Postbiotic effects of Lactobacillus fermentate on intestinal health, mucosa-associated microbiota, and growth efficiency of nursery pigs challenged with F18+Escherichia coli.

This study determined the supplemental effects of Lactobacillus fermentate (LBF, Adare Biome, France) on intestinal health and prevention of postweaning diarrhea caused by F18+Escherichia coli in nursery pigs. Sixty-four weaned pigs (6.6 ± 0.7 kg body weight) were allotted in a randomized complete block design to four treatments: NC: no challenge/no supplement; PC: E. coli challenge/no supplement; AGP: E. coli challenge/bacitracin (30 g/t feed); and PBT: E. coli challenge/LBF (2 kg/t feed). Bacitracin methylene disalicylate (BMD) was used as a source of bacitracin. On day 7, challenged groups were orally inoculated with F18+E. coli (2.4 × 1010 CFU), whereas NC received sterile saline solution. Growth performance was analyzed weekly, and pigs were euthanized at the end of 28 d feeding to analyze intestinal health. Data were analyzed using the Mixed procedure of SAS 9.4. During the post-challenge period, PC tended to decrease (P = 0.067) average daily gain (ADG) when compared with NC, whereas AGP increased (P < 0.05) when compared with PC; PBT tended to increase (P = 0.081) ADG when compared with PC. The PC increased fecal score (P < 0.05) during day 7 to 14 when compared with NC, whereas AGP decreased it (P < 0.05) during day 14 to 21 when compared with PC. The PC increased (P < 0.05) protein carbonyl, crypt cell proliferation, and the relative abundance of Helicobacter rodentium when compared with NC. However, AGP decreased (P < 0.05) crypt cell proliferation and H. rodentium and increased (P < 0.05) villus height, Bifidobacterium boum, Pelomonas spp., and Microbacterium ginsengisoli when compared with PC. The PBT reduced (P < 0.05) crypt cell proliferation and H. rodentium and increased (P < 0.05) Lactobacillus salivarius and Propionibacterium acnes when compared with PC. At the genus level, AGP and PBT increased (P < 0.05) the alpha diversity of jejunal mucosa-associated microbiota in pigs estimated with Chao1 richness estimator when compared with PC. Collectively, F18+E. coli reduced growth performance by adversely affecting microbiota and intestinal health. The LBF and BMD improved growth performance, and it was related to the enhanced intestinal health and increased diversity and abundance of beneficial microbiota in pigs challenged with F18+E. coli.

Newly weaned pigs are susceptible to multiple stressors that may lead to postweaning diarrhea, thereby causing significant economic losses in the swine industry. Enterotoxigenic Escherichia coli strains are the major agents causing diarrhea in newly weaned pigs. Subtherapeutic antibiotics have been employed by producers around the world to mitigate this issue. However, the use of antibiotics as growth promoters has become a public health concern because of microbial resistance. This study used Lactobacillus fermentate (LBF) as a postbiotic to help maintain healthy microbiota on the intestinal mucosa and to prevent postweaning diarrhea caused by E. coli F18+. Therefore, the aim of this study was to evaluate the effects of dietary supplementation of LBF on intestinal microbiota, intestinal health, and prevention of postweaning diarrhea caused by a challenge with E. coli F18+ in newly weaned pigs. Our model confirmed that the E. coli F18+ reduced growth performance by causing diarrhea, disruption of the microbiota composition, and increased immune response and oxidative stress in the small intestine of newly weaned pigs. Lactobacillus fermentate improved growth performance, and it was related to enhanced intestinal health and increased microbiota diversity in E. coli F18+-challenged pigs.

Effects of dietary bacitracin or Bacillus subtilis on the woody breast myopathy-associated gut microbiome of Eimeria spp. challenged and unchallenged broilers.

Study suggested that dysbiosis of the gut microbiota may affect the etiology of woody breast (WB). In the current study, the cecal microbiota and WB in chickens fed three different diets were investigated. A total of 504 male chicks were used in a randomized complete block design with a 3 (Diet) × 2 (Challenge) factorial arrangement of treatments with 6 replicates per treatment, 6 treatments per block, and 14 birds per treatment. The experimental diets were a control diet (corn-soybean meal basal diet), an antibiotic diet (basal diet + 6.075 mg bacitracin/kg feed), and a probiotic diet (basal diet + 2.2 × 108 CFU Bacillus subtilis PB6/kg feed). On d 14, birds that were assigned to the challenge treatment received a 20 × live cocci vaccine. On d 41, breast muscle hardness in live birds was palpated and grouped into normal (NB) and WB phenotypes. Cecal contents were collected and their bacterial compositions were analyzed and compared. The genomic DNA of the cecal contents was extracted and the V3 and V4 regions of 16S rRNA gene were amplified and sequenced via an Illumina MiSeq platform. There were no differences (P > 0.05) in Shannon and Chao 1 indexes between the challenges, diets, and phenotypes (NB vs. WB). However, there was a difference (P = 0.001) in the beta diversity of the samples between the challenged and nonchallenged groups. Relative bacterial abundance differed (false discovery rate, FDR < 0.05) between the challenge treatments, but there were no significant differences (FDR > 0.05) among the three diets or two phenotypes. Predicted energy metabolism, nucleotide metabolism, and amino acid and coenzyme biosynthesis activities only differed (q-value < 0.05) between challenged and nonchallenged groups. The cocci challenge altered the gut microbial composition on Butyricicoccus pullicaecorum, Sporobacter termitidis, and Subdoligranulum variabile, but the dietary antibiotic and probiotic treatments did not impact gut microbial composition. No strong association was found between WB myopathy and gut microbial composition in this study.

Therapeutic effect of heat-killed Lactobacillus plantarum L-137 on the gut health and growth of broilers.

Although some studies on the effects of para-probiotics on the immune system and intestinal health have been conducted independently of research on antibiotics ass growth promoters. This study investigated the effects of heat-killed Lactobacillus plantarumL-137 (L-137) and antibiotics as preventive and/or therapeutic substances for broilers against subclinical necrotic enteritis caused by Clostridium perfringens (CP). In total, 300 1-day-old broilers (46.13 ± 1.38 g) were randomly stocked at 10 birds pen-1 in five replicates and divided into six groups, namely T1 and T2, positive and negative control of CP challenge; T3 and T4, prevention with basal diet plus 10 and 50 mg/kg L-137; T5 and T6, prevention and treatment with basal diet plus 50 mg/kg of L-137 and bacitracin at 50 ppm, respectively. Broilers administered L-137 in T4, T5 and bacitracin in T6 showed an improved (p < 0.05) villus height/crypt depth ratio than control groups, suggesting that it might significantly boost growth performance. In contrast to bacitracin, a high dosage of L-137 significantly increased (p < 0.05) the spleen index value and the cytokine levels, as well as the expression of intestinal ß-defensin genes on day 28. During the 42-day production period, broilers in T4 and T5 showed a significantly enhanced (p < 0.05) expression of cytokines, AvBD-1 and AvBD-7 on day 42 compared to the control and bacitracin groups. In particular, broilers given the L-137 diets demonstrated no cumulative mortality following CP exposure, compared to a 2% mortality in T6. Our findings provide insight into eco-friendly alternatives to antibiotics for maximizing growth performance, feed efficiency and long-term disease protection in chickens; however, this has to be proven in larger-scale commercial experiments.

Dietary inulin supplementation modulates the composition and activities of carbohydrate-metabolizing organisms in the cecal microbiota of broiler chickens.

Inulin is a highly effective prebiotic and an attractive alternative to antibiotic growth promoters for increasing production and maintaining health in chickens. However, how inulin elicits its effects on members of the intestinal microbiota is unknown, even though their importance for energy metabolism and the health of chickens is well documented. A combination of 16S rRNA Illumina sequencing and transcriptomic analysis was used to investigate the effects of supplementing a corn-based basal diet with 1, 2, or 4% inulin or 400 ppm bacitracin on the composition, diversity and activities of carbohydrate-metabolizing organisms (CMOs) in the cecal microbiota of broiler chickens. We found that members of Bacteroides were the most abundant non-starch degrading CMOs, contributing 43.6-52.1% of total glycoside hydrolase genes and 34.6-47.1% activity to the meta-transcriptomes of chickens in the different dietary groups, although members of Parabacteroides, Prevotella, Alistipes, Clostridium, Barnesiella, Blastocystis, Faecalibacterium and others were also actively involved. Inulin and bacitracin inclusion in the basal diet did not change significantly the composition or diversity of these CMOs. Inulin supplementation at three levels promoted the activities of Bacteroides, Prevotella and Bifidobacterium, and 2% level appears to be the most optimal dosage for bifidobacterial activity.

Effects of Vaccination Against Coccidiosis on Gut Microbiota and Immunity in Broiler Fed Bacitracin and Berry Pomace.

Feeding practices have been found to influence gut microbiota which play a major role in immunity of poultry. In the present study, changes in cecal microbiota and humoral responses resulting in the 55 ppm bacitracin (BACI), 1% each of cranberry (CP1) and wild blueberry (BP1) pomace alone or in combination (CP+BP) feeding in broiler Cobb 500 vaccinated or not against coccidiosis were investigated. In the non-vaccinated group, no significant treatment effects were observed on performance parameters. Vaccination significantly affected bird's performance parameters particularly during the growing phase from 10 to 20 days of age. In general, the prevalence of coccidiosis and necrotic enteritis (NE) was reduced by vaccination (P < 0.05). BACI-treated birds showed low intestinal lesion scores, and both CP1 and BP1 feed supplementations reduced Eimeria acervulina and Clostridium perfringens incidences similar to BACI. Vaccination induced change in serum enzymes, minerals, and lipid levels in 21-day old birds while, levels of triglyceride (TRIG) and non-esterified fatty acids (NEFA) were higher (P < 0.05) in CP1 treated non-vaccinated group than in the control. The levels of NEFA were lower in BACI- and CP1-fed birds than in the control in non-vaccinated day 28 old birds. The highest levels of all estimated three immunoglobulins (IgY, IgM, and IgA) were found in the vaccinated birds. Metagenomics analysis of the cecal bacterial community in 21-day old birds showed the presence of Firmicutes (90%), Proteobacteria (5%), Actinobacteria (2%), and Bacteroidetes (2%). In the vaccinated group, an effect of BACI was noted on Proteobacteria (P = 0.03). Vaccination and/or dietary treatments influenced the population of Lactobacillaceae, Enterobacteriaceae, Clostridiaceae, and Streptococcaceae which were among the most abundant families. Overall, this study revealed that besides their beneficial effects on performance, alike bacitracin, berry pomaces in poultry feed have profound impacts on the chicken cecal microbiota and blood metabolites that could be influenced by vaccination against coccidiosis.

Probiotic and synbiotic in broiler diet: performance and Enterobacteriaceae / [Probiótico e simbiótico em ração para frangos: desempenho e Enterobacteriaceae]

The objective of this study was to evaluate the effects of probiotics and synbiotics on the performance and Enterobacteriaceae count of broiler chickens. A total of 640 one-day-old male broiler chicks were distributed in a completely randomized design with four treatments and eight replicates with 20 birds each. The treatments were: ration with performance enhancer (zinc bacitracin; positive control); ration without performance enhancer and probiotic/synbiotic (negative control); ration with probiotics; and ration with synbiotics. At 35 days, five birds from each treatment were euthanized and intestinal contents were harvested for determining the Enterobacteriaceae count. The performance data and average colony-forming units (CFUs) transformed as log CFU/g were subjected to analysis of variance and Tukey's test. The effects of probiotics and synbiotics were observed in the initial phase, with supplemented birds exhibiting comparable weight gain to those supplemented with bacitracin. No effect of the treatment on broiler performance was observed after 42 days. The enterobacterial count was comparable among all experimental treatments. Supplementation with probiotics and synbiotics did not compromise the performance of broilers and did not alter the Enterobacteriaceae count.(AU)

Objetivou-se avaliar o efeito do probiótico e do simbiótico sobre o desempenho e a contagem de Enterobacteriaceae em frangos. Foram utilizados 640 pintos de corte, machos, de um dia de idade, distribuídos em delineamento inteiramente ao acaso, com quatro tratamentos, oito repetições com 20 aves cada. Os tratamentos foram: ração com melhorador de desempenho (bacitracina de zinco) (controle positivo); ração sem melhorador de desempenho e sem probiótico/simbiótico (controle negativo); ração com probiótico e ração com simbiótico. Aos 35 dias, cinco aves por tratamento foram eutanasiadas para retirada de conteúdo intestinal e determinação de Enterobacteriaceae. As médias das unidades formadoras de colônias, transformadas em log/UFC/g, e de desempenho foram submetidas à análise de variância e comparadas pelo teste Tukey. Foi observado efeito do probiótico e do simbiótico na fase inicial, sendo que aves apresentaram os mesmos resultados de ganho de peso e de peso corporal que o grupo de aves alimentado com bacitracina. Aos 42 dias, não houve efeito dos tratamentos sobre o desempenho. Aves que não receberam nenhum aditivo não apresentaram maior contagem de enterobactérias, sendo semelhantes aos demais tratamentos. A adição do probiótico e do simbiótico não compromete o desempenho dos frangos e não altera a contagem de Enterobacteriaceae.(AU)

Effects of cLFchimera peptide on intestinal morphology, integrity, microbiota, and immune cells in broiler chickens challenged with necrotic enteritis.

Three hundred and sixty 1-day-old male broiler chicks were randomly allocated to 4 treatments of 6 replicates to evaluate the effects of cLFchimera, a recombinant antimicrobial peptide (AMP), on gut health attributes of broiler chickens under necrotic enteritis (NE) challenge. Treatments were as follows: (T1) unchallenged group fed with corn-soybean meal (CSM) without NE challenge and additives (NC); (T2) group fed with CSM and challenged with NE without any additives (PC); (T3) PC group supplemented with 20 mg cLFchimera/kg diet (AMP); (T4) PC group supplemented with 45 mg antibiotic (bacitracin methylene disalicylate)/kg diet (antibiotic). Birds were sampled for villi morphology, ileal microbiota, and jejunal gene expression of cytokines, tight junctions proteins, and mucin. Results showed that AMP ameliorated NE-related intestinal lesions, reduced mortality, and rehabilitated jejunal villi morphology in NE challenged birds. While the antibiotic non-selectively reduced the count of bacteria, AMP restored microflora balance in the ileum of challenged birds. cLFchimera regulated the expression of cytokines, junctional proteins, and mucin transcripts in the jejunum of NE challenged birds. In conclusion, cLFchimera can be a reliable candidate to substitute growth promoter antibiotics, while more research is required to unveil the exact mode of action of this synthetic peptide.

Dietary Mannan-oligosaccharides potentiate the beneficial effects of Bifidobacterium bifidum in broiler chicken.

This study investigated the effects of dietary Bifidobacterium bifidum (BFD) and mannan-oligosaccharide (MOS), as a synbiotic, on the production performance, gut microbiology, serum biochemistry, antioxidant profile and health indices of broiler chicken. Six dietary treatments were T1 (negative control), T2 (positive control-20 mg antibiotic BMD kg-1 diet; BMD: bacitracin methylene disalicylate), T3 (0·1% MOS + 106  CFU BFD per g feed), T4 (0·1% MOS + 107  CFU BFD per g feed), T5 (0·2% MOS + 106  CFU BFD per g feed) and T6 (0·2% MOS + 107  CFU BFD per g feed). Significantly (P < 0·01) better growth performance and efficiency was observed in birds supplemented with 0·2% MOS along with 106  CFU BFD per g of feed compared to BMD and control birds. Supplementation with 0·2% MOS along with either 106 or 107  CFU BFD per g feed reduced (P < 0·01) the gut coliform, Escherichia coli, total plate count, and Clostridium perfringens count and increased the Lactobacillus and Bifidobacterium count. Significantly (P < 0·01) higher serum and liver antioxidant enzyme pool, serum HDL cholesterol and lower serum glucose, triglyceride, total cholesterol, cardiac risk ratio, atherogenic coefficient and atherogenic index of plasma were observed in birds supplemented with 0·2% MOS along with 106  CFU BFD per g of feed compared to control or BMD supplemented birds. Better production performance, gut microbial composition, serum biochemistry, antioxidant profile and health indices were depicted by broiler chicken supplemented with 0·2% MOS and 106  CFU BFD per g of feed.

Thermal burns of the spectacle associated with supplementary heating in native New Zealand geckos.

Case history: Gradual onset of ocular opacity was observed in three gold-striped geckos (Woodworthia chrysosiretica), and five Pacific geckos (Dactylocnemis pacificus) held in two adjacent terrariums in a zoological institution located in the North Island of New Zealand. Ultraviolet light and heat had been provided for the previous 3-4 years by a fluorescent bulb, but in the last 4 weeks of winter a ceramic heat bulb had been added, situated 10 cm above the upper mesh of the cageClinical findings: All eight geckos presented with mostly bilateral lesions of varying severity confined to the central or upper quadrant of the spectacles. These lesions ranged from variable areas of opacity within the stroma of the spectacle to similarly distributed ulcers of the surface epithelium of both spectacles. The spectacle lesions in the Pacific geckos responded well to treatment with topical combined antimicrobial therapy, within 18-29 days. The gold-striped geckos suffered complications including dysecdysis, severe spectacle ulceration and perforation, mycotic spectaculitis, and widespread mycotic dermatitis resulting in death or leading to euthanasia.Pathological findings: In the three gold-striped geckos, there were extensive areas of deep ulceration and replacement of the spectacle with a thick serocellular crust containing large numbers of fungal elements. The affected areas of the stroma were expanded by large deposits of proteinaceous and mucinous material, pyknotic cellular debris and moderate numbers of heterophils and macrophages as well as infiltrating fungal hyphae.Diagnosis: Mycotic spectaculitis with ulceration and perforation, and disseminated mycotic dermatitis likely secondary to thermal burns.Clinical relevance: This is the first report of thermal burns of the spectacle in any reptile. There was species variation in the burn severity with gold-striped geckos showing more severe lesions, possibly due to a mix of behavioural and anatomical factors. The thermal burns to the spectacles in three cases were complicated by delayed healing, perforation, dysecdysis and severe mycotic infection.

Changes in cecum microbial community in response to total sulfur amino acid (TSAA: DL-methionine) in antibiotic-free and supplemented poultry birds.

The effect of essential total sulfur amino acids (TSAA) like methionine and cysteine on the cecal microbiome of broilers was investigated at 2 different time points (days 21 and 42) of broiler rearing. A total of 360-day-old Cobb male broiler chicks were randomly distributed to 6 dietary treatments in a 2 × 3 factorial arrangement, with 2 levels of antibiotic growth promoters (AGP: 0 and 0.05%) and 3 levels of TSAA (DL-methionine) either for starter (0.7, 0.8, and 0.9%) or finisher chicks (0.52, 0.62, and 0.72%), labeled as diets 1 to 6. Cecal digesta from each replicate (n = 10) were sampled on days 21 and 42. DNA was extracted for the amplification of the V4 region of bacterial 16S rRNA genes and subjected to Illumina sequencing. Bioinformatic analyses were performed using QIIME, Mothur, and ad hoc tools and functional profiles of the inferred metagenome were analyzed using PICRUST. Statistical difference was determined by 2-way ANOVA and PERMANOVA. Clustering of cecal communities using PCoA showed clear separation of microbial communities based on age (P < 0.05) of birds and between low and medium/ high levels of TSAA (DL-methionine). At day 21, bacterial richness and diversity were higher than at day 42 where Clostridium cluster XI and Lactobacillus were found most abundant. No variability in taxonomic richness at the genus level was observed with AGP and DL-methionine supplementation. Interbird variation for richness was greater at day 42 compared to day 21. The mean fold difference of richness was greater (1.5 mean fold) with diets 1 and 6, suggesting interactive effects of AGP and TSAA (DL-methionine) in the diet. KEGG function profiles calculated by PICRUST suggest that the cecal microbiome increased glycolysis and energy generation correlated with increased dietary TSAA (DL-methionine) supplementation levels during the late broiler growth period (day 42). This study increases our knowledge of microbial dynamics and functions that are relevant to host nutrition and performance that may help us tailoring alternative strategies for raising poultry birds under antibiotic-free conditions.

Effect of Bacillus subtilis DSM 32315 on the intestinal structural integrity and growth performance of broiler chickens under necrotic enteritis challenge.

The effect of dietary inclusion of Bacillus subtilis DSM 32315 on the intestinal health and growth performance of Cobb 500 male broilers subjected to a Clostridium perfringens-induced necrotic enteritis (NE) challenge was determined in 2 experiments. In experiment 1, chicks were randomly assigned to 4 treatments of 10 replicate/treatment. In experiment 2, chicks were randomly assigned to 4 treatments of 12 replicates/treatment. The experimental treatments were non-infected, non-supplemented control, infected, non-supplemented control (IC), infected + Bacillus subtilis DSM 32315 (B. subtilis DSM 32315), infected + bacitracin methylene disalicylate (BMD). In both experiments, NE was induced by oral inoculation of toxin producing C. perfringens on 3 consecutive days between 17 and 20 D of age, following exposure of birds to pre-disposing conditions. At day 28 (experiment 1), broilers fed diets with B. subtilis DSM 32315 exhibited a significantly higher body weight, lower mortality, and intestinal NE lesion score, compared to the IC treatment. At day 42 (experiment 2), B. subtilis DSM 32315 supplementation significantly improved BW, feed conversion ratio, production efficiency factor, NE lesion score, and mortality, compared to IC treatment. The effect of B. subtilis DSM 32315 on intestinal integrity of NE challenged chickens was evaluated with histomorphometry. A significantly shallower crypt depth and higher villus height to crypt depth ratio were observed in the mid-intestine of birds belonging to the B. subtilis DSM 32315 group, compared to the IC group. Furthermore, B. subtilis DSM 32315 supplementation significantly reduced the enteritis index associated with NE. In both experiments, the effect of B. subtilis DSM 32315 on the phenotypic measurements of NE and performance was comparable to the effect observed with BMD supplementation. In conclusion, supplementation of the direct fed microbial strain B. subtilis DSM 32315 can ameliorate the pathology and performance detriments associated with NE.

Metabolic Engineering of Main Transcription Factors in Carbon, Nitrogen, and Phosphorus Metabolisms for Enhanced Production of Bacitracin in Bacillus licheniformis.

Primary metabolism plays a key role in the synthesis of secondary metabolite. In this study, the main transcription factors in carbon, nitrogen, and phosphorus metabolisms (CcpA, CcpC, CcpN, CodY, TnrA, GlnR, and PhoP) were engineered to improve bacitracin yield in Bacillus licheniformis DW2, an industrial strain for bacitracin production. First, our results demonstrated that deletions of ccpC and ccpN improved ATP and NADPH supplies, and the bacitracin yields were respectively increased by 14.02% and 16.06% compared with that of DW2, while it was decreased significantly in ccpA deficient strain DW2ΔccpA. Second, excessive branched chain amino acids (BCAAs) were accumulated in codY, tnrA, and glnR deletion strains DW2ΔcodY, DW2ΔtnrA, and DW2ΔglnR, which resulted in the nitrogen catabolite repressions and reductions of bacitracin yields. Moreover, overexpression of these regulators improved intracellular BCAA supplies, and further enhanced bacitracin yields by 14.17%, 12.98%, and 16.20%, respectively. Furthermore, our results confirmed that phosphate addition reduced bacitracin synthesis capability, and bacitracin yield was improved by 15.71% in gene phop deletion strain. On the contrary, overexpression of PhoP led to a 19.40% decrease of bacitracin yield. Finally, a combinatorial engineering of these above metabolic manipulations was applied, and bacitracin yield produced by the final strain DW2-CNCTGP (Simultaneously deleting ccpC, ccpN, phop and overexpressing glnR, codY, and tnrA in DW2) reached 1014.38 U/mL, increased by 35.72% compared to DW2, and this yield was the highest bacitracin yield currently reported. Taken together, this study implied that metabolic engineering of carbon, nitrogen, and phosphorus metabolism regulators is an efficient strategy to enhance bacitracin production, and provided a promising B. licheniformis strain for industrial production of bacitracin.

Experience in patch testing: A 6-year retrospective review from a single academic allergy practice.

BACKGROUND: Patch testing is the "gold standard" to identify culprit allergen(s) causing allergic contact dermatitis (ACD), but there are limited studies of patch testing from allergy practice settings. OBJECTIVE: We sought to explore patch test findings in a large academic allergy practice, including patch testing results, history of atopy, location of dermatitis, and referral source. We also wanted to determine whether patch testing using an extended panel, such as the North American screening series, compared with a limited series, such as the Thin-Layer Rapid-Use Epicutaneous (T.R.U.E.) Test, increased the sensitivity. METHODS: A retrospective chart review was conducted of patients referred for patch testing over a 6-year period. RESULTS: A total of 585 patients (mean age 48.7 years, 71.6 % female) underwent patch testing over the 6-year period, of which 369 (63%) had a positive test. Of those who tested positive, 202 (55%) reported a history of atopy. The extremities were the most commonly involved site, followed by the head/neck and trunk. The 5 most common positive allergens were nickel sulfate, gold sodium thiosulfate, methylchloroisothiazolinone, thimerosal, and bacitracin. Three hundred fourteen (53.6%) patients were positive to at least 1 allergen on TRUE testing. Extended screening series identified an additional 10.8% of patients with positive tests who were negative to T.R.U.E. test allergens. CONCLUSION: Patch testing is a valuable diagnostic tool for the practicing allergist and provides early identification of culprit allergens in ACD. Performing an extended screening series such as the North American Contact Dermatitis Group (NACDG) or supplemental panel of allergens increased sensitivity when compared with a limited series.

Evaluation of the carryover effect of antibiotic, bee pollen and propolis on growth performance, carcass traits and splenic and hepatic histology of growing rabbits.

Sixty-four nulliparous female rabbits were distributed among eight groups (eight animals/group). Group one was the unsupplemented control group; the other seven groups were supplemented with zinc bacitracin (ZnB) at 100 mg, or bee pollen (BP) and/or propolis (Pro) at 150 and 300 mg in a capsulated form, three times a week, day after day, continuously all over the experimental period. The experiment was run for eight parties; at each parity, 28 kids of each doe group (a total of 224 rabbits) were divided into two subgroups weaned, respectively, at 24 and 30 days of age. Thus, for each parity, there were 16 groups (eight does treatments × two weaning age, 14 rabbits per group). The growing rabbits fed the standard diets without supplements. The growth performance, the carcass traits, the liver and the spleen histology of rabbits were checked up to 90 days of age to find possible carryover effects of the supplements. The supplements had no significant effect on most of the growth performance at 90 days of age, but BP150 and BP+Pro300 increased the growth rate in comparison with ZnB group. The liver weight in the control, BP300 and Pro300 groups was higher than the ZnB one. The spleen weight was higher in the groups ZnB, BP150, Pro300 and BP+Pro300, followed by the control, BP300 and BP+Pro150 and thus Pro150. The heart % in the BP150 and Pro300 groups was higher than ZnB and BP+Pro150 groups. A lymphoid hyperplasia of splenic white pulp was observed in the BP+Pro groups, while propolis alone showed a mild activation of lymphobiosis. The Pro and BP groups showed the same picture of the control group exhibiting a hydropic degeneration of mostly hepatic cells, while the ZnB group exhibited adverse effect on the bile ducts featuring portal periductal inflammatory cells infiltration with epithelial hyperplasia reflecting chronic cholangitis.

Bee pollen and propolis as dietary supplements for rabbit: Effect on reproductive performance of does and on immunological response of does and their offspring.

To evaluate the effect of bee pollen (BP) and/or propolis (Pro) supplementation on rabbit does, 64 nulliparous NZW rabbits does were distributed among eight groups (eight animals/group). One unsupplemented group was the control; the other seven groups were supplemented, respectively, with zinc bacitracin (ZnB) at 100 mg, BP at 150 and 300 mg, Pro at 150 and 300 mg, BP+Pro at 150 and 300 mg of each three times/week, day after day continuously along eight parities. The BP300, Pro300 and BP+Pro150 groups had higher body weight of litter at birth and number of kids born alive. The BP supplementation at 150 mg increased plasma total protein and albumin than the control group. The BP or Pro at 150 mg decreased plasma T3 than the other groups except for BP+Pro150. The ZnB group had significantly greater T3 /T4 ratio compared to BP, Pro and BP+Pro at 150 mg. The BP+Pro150 group had less ALT than the control; BP300 and Pro 300 mg resulted in lower plasma AST than the groups Pro150 with or without BP and the control group. The plasma alkaline phosphatase of BP at 150 or 300 mg and BP+Pro150 was significantly greater than that of the Pro150 group. The BP+Pro300 group had higher WBCs than the other groups. In contrast, the lymphocytes were greater in the Pro and BP+Pro300 groups than in BP, Pro and BP+Pro at 150 mg. The groups supplemented with BP and BP+Pro at 150 and 300 mg had significantly greater SRBCs of doe rabbits and their offspring compared to the control and the ZnB group. The BP at 300 mg increased the serum albumin and α1 -globulin than the control group. The Pro300 group had greater serum α2 -globulin and ß-globulin than the control group. The total globulin was significantly greater for the 300 mg propolis-supplemented groups than the control.