Corrigendum to "Genetic diversity and iron metabolism of Staphylococcus hominis isolates originating from bovine quarter milk, rectal feces, and teat apices" (J. Dairy Sci. 105:9995-10006).

Staphylococcus chromogenes TA showed significantly lower growth under iron-deprived conditions, and adding an iron supplement (lactoferrin or ferritin) resulted in no improvement in growth; in contrast, growth of S. chromogenes IM was significantly recovered with ferritin iron supplementation. OnlyStaphylococcus hominis strains originating from quarter milk were able to significantly utilize ferritin as an iron source to reverse the growth inhibition caused by chelating agent 2,2'-bipyridyl in varying degrees. Both S. chromogenes strains (IM and TA) and all S. hominis strains were unable to significantly use lactoferrin as an iron source for growth recovery.

Genetic diversity and iron metabolism of Staphylococcus hominis isolates originating from bovine quarter milk, rectal feces, and teat apices.

Staphylococcus hominis, a member of the non-aureus staphylococci (NAS) group, is part of the human and animal microbiota. Although it has been isolated from multiple bovine-associated habitats, its relevance as a cause of bovine mastitis is currently not well described. To successfully colonize and proliferate in the bovine mammary gland, a bacterial species must be able to acquire iron from host iron-binding proteins. The aims of this study were (1) to assess the genetic diversity of S. hominis isolated from bovine quarter milk, rectal feces, and teat apices, and (2) to investigate the capacity of bovine S. hominis isolates belonging to these different habitats to utilize ferritin and lactoferrin as iron sources. To expand on an available collection of bovine S. hominis isolates (2 from quarter milk, 8 from rectal feces, and 19 from teat apices) from one commercial dairy herd, a subsequent single cross-sectional quarter milk sampling (n = 360) was performed on all lactating cows (n = 90) of the same herd. In total, 514 NAS isolates were recovered and identified by MALDI-TOF mass spectrometry; the 6 most prevalent NAS species were S. cohnii (33.9%), S. sciuri (16.7%), S. haemolyticus (16.3%), S. xylosus (9.6%), S. equorum (9.4%), and S. hominis (3.5%). A random amplified polymorphic DNA (RAPD) analysis was performed on 46 S. hominis isolates (19 from quarter milk, 8 from rectal feces, and 19 from teat apices). Eighteen distinct RAPD fingerprint groups were distinguished although we were unable to detect the presence of the same RAPD type in all 3 habitats. One S. hominis isolate of a distinct RAPD type unique to a specific habitat (8 from quarter milk, 3 from rectal feces, and 4 from teat apices) along with the quality control strain Staphylococcus aureus ATCC 25923 and 2 well-studied Staphylococcus chromogenes isolates ("IM" and "TA") were included in the phenotypical iron test. All isolates were grown in 4 types of media: iron-rich tryptic soy broth, iron-rich tryptic soy broth deferrated by 2,2'-bipyridyl, and deferrated tryptic soy broth supplemented with human recombinant lactoferrin or equine spleen-derived ferritin. The growth of the different strains was modified by the medium in which they were grown. Staphylococcus chromogenes TA showed significantly lower growth under iron-deprived conditions, and adding an iron supplement (lactoferrin or ferritin) resulted in no improvement in growth; in contrast, growth of S. chromogenes IM was significantly recovered with iron supplementation. Staphylococcus hominis strains from all 3 habitats were able to significantly utilize ferritin but not lactoferrin as an iron source to reverse the growth inhibition, in varying degrees, caused by the chelating agent 2,2'-bipyridyl.

Short communication: Effect of freezer storage time and thawing method on the recovery of Mycoplasma bovis from bovine colostrum.

Mycoplasma bovis is an important cause of mastitis in dairy cattle, and pneumonia, arthritis, and otitis in calves. Milk and colostrum are considered important sources of infection for calves. knowledge on the effect of on-farm freezing (-18°C) and thawing methods on the recovery of M. bovis from colostrum samples is missing. In this study, 2 separate experiments were performed. The first experiment consisted of a longitudinal study examining the survival [as measured by log(10) reduction] of 2 M. bovis strains in frozen colostrum over 14 wk. The second experiment examined the effect of different thawing temperatures (45 and 20°C), thawing frequencies (once or twice), and initial colostrum titer (104 or 106 cfu/mL) on M. bovis survival. A single freeze-thaw cycle led to an approximate 1 log reduction of M. bovis titer, independent of the thawing temperature. Freezing for 14 wk did not significantly further reduce the titer of bacteria compared with freezing for 2 wk. A second freeze-thaw cycle further reduced the M. bovis count by approximately 0.5 log compared with a single freeze-thaw cycle. Thawing temperature and initial bacterial concentration did not significantly affect M. bovis reduction. In conclusion, storage of colostrum samples in the freezer at -18°C during epidemiological studies, herd monitoring, or test and cull programs will probably have little influence on qualitative bacteriological test results for M. bovis. The epidemiological or clinical relevance of an approximate 1 log reduction of M. bovis in colostrum is currently unclear.

Successful treatment of Batrachochytrium salamandrivorans infections in salamanders requires synergy between voriconazole, polymyxin E and temperature.

Chytridiomycosis caused by the chytrid fungus Batrachochytrium salamandrivorans (Bsal) poses a serious threat to urodelan diversity worldwide. Antimycotic treatment of this disease using protocols developed for the related fungus Batrachochytrium dendrobatidis (Bd), results in therapeutic failure. Here, we reveal that this therapeutic failure is partly due to different minimum inhibitory concentrations (MICs) of antimycotics against Bsal and Bd. In vitro growth inhibition of Bsal occurs after exposure to voriconazole, polymyxin E, itraconazole and terbinafine but not to florfenicol. Synergistic effects between polymyxin E and voriconazole or itraconazole significantly decreased the combined MICs necessary to inhibit Bsal growth. Topical treatment of infected fire salamanders (Salamandra salamandra), with voriconazole or itraconazole alone (12.5 µg/ml and 0.6 µg/ml respectively) or in combination with polymyxin E (2000 IU/ml) at an ambient temperature of 15 °C during 10 days decreased fungal loads but did not clear Bsal infections. However, topical treatment of Bsal infected animals with a combination of polymyxin E (2000 IU/ml) and voriconazole (12.5 µg/ml) at an ambient temperature of 20 °C resulted in clearance of Bsal infections. This treatment protocol was validated in 12 fire salamanders infected with Bsal during a field outbreak and resulted in clearance of infection in all animals.

Chronic exposure to the mycotoxin T-2 promotes oral absorption of chlortetracycline in pigs.

The aim of this study was to investigate whether T-2 toxin, a potent Fusarium mycotoxin, affects the oral absorption of the antibiotic chlortetracycline in pigs. Animals were allocated to blank feed without T-2 toxin (controls), feed containing 111 µg T-2/kg feed, T-2-contaminated feed supplemented with a yeast-derived feed additive, or blank feed supplemented solely with the feed additive, respectively. After 21 days, an intragastric bolus of chlortetracycline was given to assess potential alterations in the pharmacokinetics of this commonly used antibiotic. A significantly higher area under the plasma concentration-time curve and maximal plasma concentration of chlortetracycline was observed after intake of T-2-contaminated feed compared with control. Thus, exposure to T-2-contaminated feed can influence the oral bioavailability of chlortetracycline. This effect could have consequences for the withdrawal time of the drug and the occurrence of undesirable residues in edible tissues.

Evaluation of three intervention strategies to reduce the transmission of Salmonella Typhimurium in pigs.

Despite current control measures, Salmonella in pigs remains a major public health concern. In this in vivo study, the effect of three intervention strategies on Salmonella Typhimurium transmission in pigs was evaluated. The first intervention was feed supplemented with coated calcium-butyrate (group A); the second comprised oral vaccination with a double-attenuated Salmonella Typhimurium strain (group B), and the third was acidification of drinking water with a mixture of organic acids (group C). After challenge at 8 weeks of age, animals were individually sampled for 6 weeks (blood once per week; faeces twice per week) and then were euthanased at 14 weeks of age. Post-mortem ileum, caecum, ileocaecal lymph nodes, and tonsils were sampled, along with ileal, caecal and rectal contents, and tested for the presence of Salmonella spp. Transmission was quantified by calculating an 'adjusted' reproduction ratio 'Ra' and its 95% confidence interval (CI). The proportion of pigs that excreted Salmonella spp. via the faeces was significantly higher in group C (58%, P<0.0001) and the positive control group (41%, P=0.03), compared to group B (15%), and the proportion in group C was also significantly higher than in group A (23%, P=0.01). Group A had the lowest proportion of positive post-mortem samples (18%), followed by group B (31%), the positive control group (41%) and group C (64%) (P<0.03). The highest transmission was seen in the positive control group and group C (Ra=+∞ with 95% CI [1.88; +∞]), followed by group B (Ra=2.61 [1.21; 9.45]) and A (Ra=1.76 [1.02; 9.01]). The results of this study suggest that vaccination and supplementation of the feed with coated calcium-butyrate limited Salmonella transmission in pigs and might be useful control measures.

In vitro sensitivity of poultry Brachyspira intermedia isolates to essential oil components and in vivo reduction of Brachyspira intermedia in rearing pullets with cinnamaldehyde feed supplementation.

Cecal enteritis due to Brachyspira infections tends to be chronic in laying hens. Limited availability of antimicrobial drugs for use in laying hens emphasizes the need for alternative control measures. A broth microdilution method was used to determine the antimicrobial susceptibility of 20 Brachyspira intermedia field isolates from laying hen flocks to components of essential oils (EO). Minimal inhibitory concentration (MIC) distributions, obtained for 8 EO components, were all monomodal. Cinnamaldehyde had the lowest MIC values (40 to 80 mg/L), followed by nerolidol, capsaicin, carvacrol, and thymol (80 to 320 mg/L), eugenol (160 to 640 mg/L), and linalool (320 to 1,280 mg/L). The MIC ranges of piperine were mostly above the test range of 1,280 mg/L. In an in vivo experiment, coated trans-cinnamaldehyde was supplemented to the feed of rearing pullets. A completely randomized experimental design with 4 treatments and 3 replicates each (replicate = group of seven 1-d-old laying hen chickens) was applied. The negative and positive controls received a conventional feed during the whole trial. The positive controls were orally inoculated on 3 consecutive days (d 22, 23, and 24) with 1 mL of 1.0 × 10(8) cfu/mL of a B. intermedia field isolate. Two treatment groups (preventive and curative), identically inoculated, received the coated trans-cinnamaldehyde-supplemented feed (500 mg/kg of trans-cinnamaldehyde), the preventive group from d 1, the curative from d 25. On d 32, ceca were collected for bacteriologic Brachyspira enumeration. The mean enumeration of Brachyspira cells was decreased (P < 0.05) in the curative treated group versus the positive control group. The in vitro results of the present study demonstrate the potential of EO components as antimicrobials against poultry Brachyspira isolates, including isolates with acquired resistance for classic antimicrobial drugs. Reduction of Brachyspira colonization in young pullets was obtained, in a curative way, in an in vivo study using feed supplemented with coated trans-cinnamaldehyde. Further studies are necessary to investigate the mode of action of the coated trans-cinnamaldehyde in reducing Brachyspira colonization of the ceca.

Developing a safe antifungal treatment protocol to eliminate Batrachochytrium dendrobatidis from amphibians.

Batrachochytrium dendrobatidis is one of the most pathogenic microorganisms affecting amphibians in both captivity and in nature. The establishment of B. dendrobatidis free, stable, amphibian captive breeding colonies is one of the emergency measures that is being taken to save threatened amphibian species from extinction. For this purpose, in vitro antifungal susceptibility testing and the development of efficient and safe treatment protocols are required. In this study, we evaluated the use of amphotericin B and voriconazole to treat chytridiomycosis in amphibians. The concentration at which the growth of five tested B. dendrobatidis strains was inhibited was 0.8 µg/ml for amphotericin B and 0.0125 µg/ml for voriconazole. To completely eliminate a mixture of sporangia and zoospores of strain IA042 required 48 h of exposure to 8 µg/ml of amphotericin B or 10 days to 1.25 µg/ml of voriconazole. Zoospores were killed within 0.5 h by 0.8 µg/ml of amphotericin B, but even after 24 h exposure to 1.25 µg/ml of voriconazole they remained viable. Amphotericin B was acutely toxic for Alytes muletensis tadpoles at 8 µg/ml, whereas toxic side effects were not noticed during a seven-day exposure to voriconazole at concentrations as high as 12.5 µg/ml. The voriconazole concentrations remained stable in water during this exposure period. On the basis of this data, experimentally inoculated postmetamorphic Alytes cisternasii were sprayed once daily for 7 days with a 1.25 µg/ml solution of voriconazole in water which eliminated the B. dendrobatidis infection from all treated animals. Finally, treatment of a naturally infected colony of poison dart frogs (Dendrobatidae) using this protocol, combined with environmental disinfection, cleared the infection from the colony.

Intestinal mucus protects Campylobacter jejuni in the ceca of colonized broiler chickens against the bactericidal effects of medium-chain fatty acids.

Campylobacter jejuni is the most common cause of bacterial-mediated diarrheal disease worldwide. Because poultry and poultry products are a major source of C. jejuni infections in humans, efforts should be taken to develop strategies to decrease Campylobacter shedding during primary production. For this purpose, the efficacy of medium-chain fatty acids (MCFA) as feed additives to control C. jejuni colonization in broiler chickens was analyzed. First, the antimicrobial activity of the MCFA caproic, caprylic, and capric acid on C. jejuni was evaluated in vitro. Minimal inhibitory concentrations were 0.25 mM for caproic and 0.5 mM for caprylic and capric acids at pH 6.0 and 4 mM for all 3 compounds at pH 7.5. Time-kill curves revealed strong bactericidal properties of the tested compounds toward C. jejuni at pH 6.0. Concentrations as low as 4 mM caprylic and capric acids and 16 mM caproic acid killed all bacteria within 24 h. Capric acid had the highest activity, with concentrations of 4 mM killing all bacteria within the hour. Together these data show a profound bactericidal, dose-dependent activity of the tested MCFA toward C. jejuni in vitro. For this reason, the effect of these 3 MCFA on C. jejuni was evaluated in vivo. The addition of any of the acids to the feed, from 3 d before euthanization, was not capable of reducing cecal Campylobacter colonization in 27-d-old broilers experimentally infected with C. jejuni at 15 d of age. Using a cecal loop model, sodium caprate was not able to reduce cecal Campylobacter counts. When time-kill curves were conducted in the presence of chick intestinal mucus, capric acid was less active against C. jejuni. At 4 mM, all bacteria were killed only after 24 h. Thus, despite the marked bactericidal effect of MCFA in vitro, supplementing these acids to the feed does not reduce cecal Campylobacter colonization in broiler chickens under the applied test conditions, probably due to the protective effect of the mucus layer.

The effect of commonly used anticoccidials and antibiotics in a subclinical necrotic enteritis model.

Necrotic enteritis poses an important health risk to broilers. The ionophore anticoccidials lasalocid, salinomycin, maduramicin, narasin and a combination of narasin and nicarbazin were tested in feed for their prophylactic effect on the incidence of necrotic enteritis in a subclinical experimental infection model that uses coccidia as a predisposing factor. In addition, drinking water medication with the antibiotics amoxicillin, tylosin and lincomycin was evaluated as curative treatment in the same experimental model. The minimal inhibitory concentrations (MICs) of all antibiotics and anticoccidials were determined in vitro against 51 Clostridium perfringens strains isolated from broilers. The strains examined appeared uniformly susceptible to lasalocid, maduramicin, narasin, salinomycin, amoxicillin and tylosin, whereas an extended frequency distribution range of MICs for lincomycin was seen, indicating acquired resistance in 36 isolates in the higher range of MICs. Nicarbazin did not inhibit the in vitro growth of the C. perfringens strains even at a concentration of 128 microg/ml. Supplementation of the diet from day 1 onwards with lasalocid, salinomycin, narasin or maduramicin led to a reduction in birds with necrotic enteritis lesions as compared with the non-medicated infected control group. A combination product of narasin and nicarbazin had no significant protective effect. Treatment with amoxicillin, lincomycin and tylosin completely stopped the development of necrotic lesions.

Control of Clostridium perfringens-induced necrotic enteritis in broilers by target-released butyric acid, fatty acids and essential oils.

The efficacy of target-released butyric acid, medium-chain fatty acids (C(6) to C(12) but mainly lauric acid) and essential oils (thymol, cinnamaldehyde, essential oil of eucalyptus) micro-encapsulated in a poly-sugar matrix to control necrotic enteritis was investigated. The minimal inhibitory concentrations of the different additives were determined in vitro, showing that lauric acid, thymol, and cinnamaldehyde are very effective in inhibiting the growth of Clostridium perfringens. The in vivo effects were studied in two trials in an experimental necrotic enteritis model in broiler chickens. In the first trial, four groups of chickens were fed a diet supplemented with butyric acid, with essential oils, with butyric acid in combination with medium-chain fatty acids, or with butyric acid in combination with medium-chain fatty acids and essential oils. In all groups except for the group receiving only butyric acid, a significant decrease in the number of birds with necrotic lesions was found compared with the infected, untreated control group. In the second trial the same products were tested but at a higher concentration. An additional group was fed a diet supplemented with only medium-chain fatty acids. In all groups except for that receiving butyric acid in combination with medium-chain fatty acids and essential oils, a significant decrease in the number of birds with necrotic lesions was found compared with the infected, untreated control group. These results suggest that butyric acid, medium-chain fatty acids and/or essential oils may contribute to the prevention of necrotic enteritis in broilers.

Voriconazole, a safe alternative for treating infections caused by the Chrysosporium anamorph of Nannizziopsis vriesii in bearded dragons (Pogona vitticeps).

Dermal and systemic infections caused by the Chrysosporium anamorph of Nannizziopsis vriesii (CANV) are highly prevalent in reptiles and may result in severe disease and high mortality. Due to the high incidence of therapeutic failures, optimizing treatment is required. We first determined in this study the minimal inhibitory concentrations (MIC) of itraconazole, voriconazole, amphotericin B and terbinafine against 32 CANV isolates. For voriconazole, amphotericin B and terbinafine a monomodal MIC distribution was seen, whereas a bimodal MIC distribution was present for itraconazole, indicating acquired resistance in one isolate. Fourteen naturally-infected bearded dragons (Pogona vitticeps), from the same owner, were treated orally with either itraconazole (5 mg/kg q24h) or voriconazole (10 mg/kg q24h). The clinical condition, drug plasma concentrations and the presence of CANV in skin samples were followed. The animals were treated until complete clearance of the fungus. The plasma concentrations of voriconazole and itraconazole exceeded the minimal inhibitory concentrations of the CANV isolates. Elimination of CANV was achieved on average after 27 and 47 days of treatment with itraconazole and voriconazole, respectively. Whereas only 2 out of 7 survived after itraconazole treatment, only a single animal died in the voriconazole treated group. In conclusion, based on a limited number of animals, voriconazole applied at a regimen of 10 mg/kg bodyweight (BW) q24h seems to be a safe and effective antimycotic drug to eliminate CANV infections in bearded dragons.

The use of tannins to control Salmonella typhimurium infections in pigs.

The aim of this study was to determine whether a hydrolysable tannin extract of sweet chestnut wood (Globatan(®)) has an inhibitory effect on Salmonella Typhimurium survival both in vitro and in vivo in pigs. In a first experiment, the minimal inhibitory concentration of Globatan(®) on 57 Salmonella Typhimurium isolates was determined. For all isolates, an MIC of 160-320 µg/ml was found. The second in vitro study revealed that Salmonella growth was strongly reduced using Globatan(®) concentrations of 25-50 µg/ml and nearly completely inhibited at a concentration of 100 µg/ml Globatan(®). In an in vivo trial, two groups of six piglets, each group receiving feed with or without the addition of Globatan(®) (3 g/kg), were orally inoculated with 10(7) colony forming units of a Salmonella Typhimurium strain. Globatan(®) had no effect on faecal excretion of Salmonella, and no differences in colonization of the intestines and internal organs were demonstrated in pigs euthanized at 4 days post-inoculation. In conclusion, the hydrolysable tannin extract used in this study showed strong action against Salmonella Typhimurium in vitro but not in vivo.

The effect of reduced treatment time and dosage of enrofloxacin on the course of respiratory disease caused by avian metapneumovirus and Ornithobacterium rhinotracheale.

A dose titration and reduced duration medication study were performed to evaluate the current enrofloxacin treatment schedule in growing turkeys experimentally infected with avian metapneumovirus and Ornithobacterium rhinotracheale. Experimental groups of 17 four-week-old turkeys were first infected with avian metapneumovirus and 3 d later with O. rhinotracheale. Enrofloxacin treatment in the drinking water was started 24 h after O. rhinotracheale inoculation. In the dose titration study, enrofloxacin doses of 5, 10, and 20 mg/kg of BW were administered for 5 successive days. In the reduced duration medication study, the following enrofloxacin regimens were compared: 25 mg/kg of BW per day on d 0 and 2; 15 mg/kg of BW per day on d 0, 2, and 4; and 10 mg/kg of BW for 5 successive days. In both studies, all enrofloxacin treatments were equally efficacious (i.e., equally capable of shortening the course of clinical disease), eliminating O. rhinotracheale from the respiratory tract and reducing gross lesions. Ornithobacterium rhinotracheale bacteria were not recovered from any of the birds on enrofloxacin-supplemented media, indicating that none of the used treatment regimens promoted the selection of bacterial clones with reduced susceptibility or resistance to this antimicrobial agent. In conclusion, none of the alternative enrofloxacin treatment regimens yielded better results than the current prescribed treatment (i.e., 10 mg/kg of BW for 5 successive days) of O. rhinotracheale infections in turkeys. However, the reduced duration of application would offer a less time-consuming and equally effective alternative.

The cereal type in feed influences Salmonella Enteritidis colonization in broilers.

A study was conducted to analyze the effect of the cereal type in the diet on the susceptibility for Salmonella Enteritidis in broilers. In the first experiment, 40 newly hatched broiler chicks were randomly assigned to 4 different experimental treatment groups. Two treatment groups were fed a maize-soybean-based diet, of which one contained 100 mg/kg of the antibiotic growth promoter zinc bacitracin. The 2 other treatment groups were fed a wheat/rye-soybean-based diet, of which one was supplemented with 100 mg/kg of zinc bacitracin. The broilers were inoculated with Salmonella Enteritidis at d 11 and killed at d 15. When the nonantibiotic- and the antibiotic-treated birds were pooled, the Salmonella colonization in the spleen and ceca of the birds fed a maize-based diet was significantly lower in comparison to colonization of spleen and ceca in the wheat/rye groups. Zinc bacitracin did not affect Salmonella colonization. In a second experiment, which was a modified repetition of the first experiment, 120 newly hatched broiler chicks were randomly assigned to 2 experimental treatments. They were fed a maize-soybean or a wheat/rye-soybean based diet, inoculated with Salmonella Enteritidis at d 11, and killed at d 15. A significantly lower Salmonella colonization was observed in the spleen, liver, and ceca of broilers given a maize-based diet in comparison to those given a wheat/rye-based diet. These data show that the cereal type in broiler feed can affect Salmonella colonization probably due to changes in intestinal health of the birds. Hence, altering the diet composition can be regarded as a simple tool to supplement other control measures against Salmonella in broilers.

Coated fatty acids alter virulence properties of Salmonella Typhimurium and decrease intestinal colonization of pigs.

Salmonella Typhimurium infections in pigs are a major source of human foodborne salmonellosis. To reduce the number of infected pigs, acidification of feed or drinking water is a common practice. The aim of the present study was to determine whether some frequently used short- (SCFA) and medium-chain fatty acids (MCFA) are able to alter virulence gene expression and to decrease Salmonella Typhimurium colonization and shedding in pigs using well established and controlled in vitro and in vivo assays. Minimal inhibitory concentrations (MIC) of 4 SCFA (formic acid, acetic acid, propionic acid and butyric acid) and 2 MCFA (caproic and caprylic acid) were determined using 54 porcine Salmonella Typhimurium field strains. MIC values increased at increasing pH-values and were two to eight times lower for MCFA than for SCFA. Expression of virulence gene fimA was significantly lower when bacteria were grown in LB-broth supplemented with sub-MIC concentrations of caproic or caprylic acid (2 mM). Expression of hilA and invasion in porcine intestinal epithelial cells was significantly lower when bacteria were grown in LB-broth containing sub-MIC concentrations of butyric acid or propionic acid (10 mM) and caproic or caprylic acid (2 mM). When given as feed supplement to pigs experimentally infected with Salmonella Typhimurium, coated butyric acid decreased the levels of faecal shedding and intestinal colonization, but had no influence on the colonization of tonsils, spleen and liver. Uncoated fatty acids, however, did not influence fecal shedding, intestinal or tonsillar colonization in pigs. In conclusion, supplementing feed with certain coated fatty acids, such as butyric acid, may help to reduce the Salmonella load in pigs.

Supplementation of coated butyric acid in the feed reduces colonization and shedding of Salmonella in poultry.

Short-chain fatty acids have been widely used as feed additives to control Salmonella in poultry. Data on the use of butyric acid in poultry are lacking. In this study, powder form and coated butyric acid were compared in their ability to reduce Salmonella colonization of ceca and internal organs shortly after infection of young chickens with Salmonella enteritidis. In the first trial, 4 groups of 25 specific pathogen free layer chickens were given feed either supplemented with powder form butyric acid, coated butyric acid, a combination of powder form and coated butyric acid (all groups received a total of 0.63 g of butyric acid/kg) or nonsupplemented feed. The specific pathogen free layer chickens were orally infected with 10(6) cfu of S. enteritidis. Coated butyric acid significantly decreased cecal colonization 3 d post-infection compared with control chickens, and powder form butyric acid had no effect. To study long-term shedding and colonization of Salmonella in broilers given coated butyric acid as feed additive (0.63 g of active product butyric acid/kg), 10 Ross broiler chickens were infected at d 5 with 10(5) cfu of S. enteritidis and housed together with 40 noninfected broilers. A control group received nonsupplemented feed. The group of broilers receiving coated butyric acid had a significantly lower number of broilers shedding Salmonella bacteria, but cecal colonization at slaughter age was equal for both groups. In conclusion, butyric acid decreases cecal colonization shortly after infection, decreases fecal shedding, and as a consequence, decreases environmental contamination by S. enteritidis-infected broilers. However, complete elimination can probably only be achieved with a combined approach using both hygienic measures and different protection measures, as the broilers still carried S. enteritidis bacteria in the ceca at slaughter age, although at enrichment level.

Microencapsulated short-chain fatty acids in feed modify colonization and invasion early after infection with Salmonella enteritidis in young chickens.

Short-chain fatty acids (SCFA) are widely used as feed additives in poultry for the control of pathogenic bacteria, such as Salmonella enteritidis. Recently, a new range of products was developed in which SCFA are encapsulated in mineral carriers, resulting in a slow release during the transport of these carriers through the intestinal tract. To test the efficacy of this type of products against early colonization after Salmonella infection in poultry, a challenge experiment with S. enteritidis was performed. Five groups of 20 chickens were given feed with no supplement or feed supplemented with acetic acid (0.24%), formic acid (0.22%), or propionic acid (0.27%) as film-coated microbeads or butyric acid (0.15%) as spray-cooled microcapsules. The 5 groups were challenged with 5 x 10(3) cfu S. enteritidis at d 5 and 6 posthatch, and samples of ceca, liver, and spleen were taken at d 8 and analyzed for the number of colony-forming units of Salmonella per gram of tissue. Feed supplementation with acetic acid, and to a lesser extent formic acid, resulted in an increase of colonization of ceca and internal organs. Birds receiving propionic acid-coated microbeads as feed supplement were colonized with Salmonella to the same extent as controls. Butyric acid-impregnated microbeads in the feed, however, resulted in a significant decrease of colonization by S. enteritidis in the ceca but not in liver and spleen.

Invasion of Salmonella enteritidis in avian intestinal epithelial cells in vitro is influenced by short-chain fatty acids.

Fermentation reactions in the caeca of chickens, the predominant place for Salmonella colonization, result in high concentrations of short-chain fatty acids (SCFA). Thus Salmonella bacteria are in close contact with SCFA during their life cycle. A study was carried out to analyse the effects of SCFA on invasion of Salmonella enteritidis in an avian intestinal epithelial cell line. Preincubation of S. enteritidis for 4 h in growth media supplemented with various concentrations of propionate or butyrate resulted in decreased invasion compared to bacteria, preincubated in nonsupplemented media, and to bacteria, preincubated in media supplemented with formate or acetate. Incubation of the S. enteritidis bacteria in media supplemented with mixtures of SCFA mimicking the in vivo caecal concentrations resulted in increased invasion compared with butyrate-exposed bacteria, but equal invasion compared with nonexposed bacteria. Increasing the butyrate concentration in these mixtures did not modify invasion compared with the original mixtures.

In vitro susceptibility of Enterococcus faecium isolated from food to growth-promoting and therapeutic antibiotics.

A total of 76 E. faecium strains, isolated at retail level from raw poultry meat, cheese, raw pork, and preparations of cheese and raw pork, were tested for their susceptibility and resistance to growth-promoting antibacterials used in animals and antibiotics used therapeutically in humans. All strains were uniformly susceptible to the growth promoters bambermycin and avilamycin. Resistance against bacitracin, virginiamycin and narasin was high among strains from poultry meat. With tylosin, a macrolide antibiotic used therapeutically and for growth promotion, resistance was mainly detected in strains originating from poultry meat, though also in some strains from pork and from pork and cheese preparations. The therapeutic antibiotic dalfopristin/quinupristin did not show full cross-resistance with the growth-promoting antibiotic virginiamycin. With dalfopristin/quinupristin two different levels of resistance were found. Only one E. faecium strain isolated from poultry was resistant to the glycopeptides avoparcin and vancomycin. Only one poultry meat strain was highly resistant to ampicillin. However, nearly all poultry meat strains showed decreased sensitivity. Only 3 out of 24 poultry strains were susceptible to minocycline, while all strains from other origins were susceptible to this tetracycline antibiotic. High-level streptomycin resistance was seen in strains of all origins, though infrequently. High-level gentamicin resistance was not found.