Sequence-Based Analysis of the Intestinal Microbiota of Sows and Their Offspring Fed Genetically Modified Maize Expressing a Truncated Form of Bacillus thuringiensis Cry1Ab Protein (Bt Maize)

The aim was to investigate transgenerational effects of feeding genetically modified (GM) maize expressing a truncated form of Bacillus thuringiensis Cry1Ab protein (Bt maize) to sows and their offspring on maternal and offspring intestinal microbiota. Sows were assigned to either non-GM or GM maize dietary treatments during gestation and lactation. At weaning, offspring were assigned within sow treatment to non-GM or GM maize diets for 115 days, as follows: (i) non-GM maize-fed sow/non-GM maize-fed offspring (non-GM/non-GM), (ii) non-GM maize-fed sow/GM maize-fed offspring (non-GM/GM), (iii) GM maize-fed sow/non-GM maize-fed offspring (GM/non-GM), and (iv) GM maize-fed sow/GM maize-fed offspring (GM/GM). Offspring of GM maize-fed sows had higher counts of fecal total anaerobes and Enterobacteriaceae at days 70 and 100 postweaning, respectively. At day 115 postweaning, GM/non-GM offspring had lower ileal Enterobacteriaceae counts than non-GM/non-GM or GM/GM offspring and lower ileal total anaerobes than pigs on the other treatments. GM maize-fed offspring also had higher ileal total anaerobe counts than non-GM maize-fed offspring, and cecal total anaerobes were lower in non-GM/GM and GM/non-GM offspring than in those from the non-GM/non-GM treatment. The only differences observed for major bacterial phyla using 16S rRNA gene sequencing were that fecal Proteobacteria were less abundant in GM maize-fed sows prior to farrowing and in offspring at weaning, with fecal Firmicutes more abundant in offspring. While other differences occurred, they were not observed consistently in offspring, were mostly encountered for low-abundance, low-frequency bacterial taxa, and were not associated with pathology. Therefore, their biological relevance is questionable. This confirms the lack of adverse effects of GM maize on the intestinal microbiota of pigs, even following transgenerational consumption.     

Diverse Tetracycline Resistance Genotypes of Megasphaera elsdenii Strains Selectively Cultured from Swine Feces

A total of 30 Megasphaera elsdenii strains, selectively isolated from the feces of organically raised swine by using Me109 M medium, and one bovine strain were analyzed for tetracycline resistance genotypic and phenotypic traits. Tetracycline-resistant strains carried tet(O), tet(W), or a tet gene mosaic of tet(O) and tet(W). M. elsdenii strains carrying tet(OWO) genes exhibited the highest tetracycline MICs (128 to >256 μg/ml), suggesting that tet(O)-tet(W) mosaic genes provide the selective advantage of greater tetracycline resistance for this species. Seven tet genotypes are now known for M. elsdenii, an archetype commensal anaerobe and model for tet gene evolution in the mammalian intestinal tract.     

Perinatal flavour learning and adaptation to being weaned: all the pig needs is smell

Perinatal flavour learning through the maternal diet is known to enhance flavour preference and acceptance of flavoured food in many species, yet still little is known about the mechanism underlying perinatal flavour learning. Previously we found positive effects of perinatal flavour learning on food intake, growth and behaviour of piglets postweaning, but no increased preference for the flavour. This suggests that flavour learning in pigs works through a reduction of weaning stress by the presence of the familiar flavour instead. The aim of this study was to investigate whether perinatal flavour learning reduces stress at weaning, and whether the effect is stronger when the familiar flavour is present in the food. Sows were offered an anethol-flavoured diet (Flavour treatment) or control diet (Control treatment) during late gestation and lactation. Flavour and Control piglets were provided with anethol either in their food (Food treatment) or in the air (Air treatment) after weaning. Preweaning and postweaning treatments did not affect food intake, preference or growth in the first two weeks postweaning but flavour treatment reduced the latency to eat (24 versus 35 hours, P = 0.02) and within-pen variation in growth (SD within-pen: 0.7 versus 1.2 kg, P<0.001). Salivary cortisol levels tended to be lower four and seven hours postweaning for Flavour piglets compared to Control piglets (4 hours: 2.5 versus 3.0 ng/ml, P = 0.05, 7 hours: 3.1 versus 3.4 ng/ml, P = 0.08). Flavour piglets played more and showed less damaging behaviours than Control piglets, indicating that the familiar flavour reduced stress around weaning. Few interaction effects were found between preweaning and postweaning treatment, and no effects of postweaning treatment. We conclude that in the newly weaned pig, perinatal flavour learning results in a reduction of stress when the familiar flavour is present, regardless of providing the flavour in the food or in the air.     

Transfer of the Pheromone-Inducible Plasmid pCF10 among Enterococcus faecalis Microorganisms Colonizing the Intestine of Mini-Pigs

A new animal model, the streptomycin-treated mini-pig, was developed in order to allow colonization of defined strains of Enterococcus faecalis in numbers sufficient to study plasmid transfer. Transfer of the pheromone-inducible pCF10 plasmid between streptomycin-resistant strains of E. faecalis OG1 was investigated in the model. The plasmid encodes resistance to tetracycline. Numbers of recipient, donor, and transconjugant bacteria were monitored by selective plating of fecal samples, and transconjugants were subsequently verified by PCR. After being ingested by the mini-pigs, the recipient strain persisted in the intestine at levels between 10⁶ and 10⁷ CFU per g of feces throughout the experiment. The donor strain, which carried different resistance markers but was otherwise chromosomally isogenic to the recipient strain, was given to the pigs 3 weeks after the recipient strain. The donor cells were initially present in high numbers (10⁶ CFU per g) in feces, but they did not persist in the intestine at detectable levels. Immediately after introduction of the donor bacteria, transconjugant cells appeared and persisted in fecal samples at levels between 10³ and 10⁴ CFU per g until the end of the experiment. These observations showed that even in the absence of selective tetracycline pressure, plasmid pCF10 was transferred from ingested E. faecalis cells to other E. faecalis organisms already present in the intestinal environment and that the plasmid subsequently persisted in the intestine.     

Interaction of CP levels in maternal and nursery diets,and its effect on performance,protein digestibility,and serum urea levels in piglets

Reduced protein levels in nursery diets have been associated with a lower risk of postweaning diarrhea, but the interaction with CP levels in maternal diet on the performance of the offspring remains unclear. The objective of this study was to determine the effect of protein content in sow gestation and piglet nursery diets on the performance of the piglets until slaughter. This was studied in a 2 × 2 factorial trial (35 sows, 209 piglets), with higher or lower (H or L) dietary CP in sow diets (168 vs 122 g CP/kg) during late gestation. A standard lactation feed was provided for all sows (160 g CP/kg). For both sow treatments, half of the litters received a higher or lower CP in the piglet nursery diet (210 vs 166 g CP/kg). This resulted in four possible treatment combinations: HH, HL, LH and LL, with sow treatment as first and piglet treatment as second letter. For each phase, all diets were iso-energetic and had a similar level of essential amino acids. P_s*p is the p-value for the interaction effect between sow and piglet treatment. In the nursery phase (3.5–9 weeks of age), a tendency toward interaction between piglet and sow treatments with feed efficiency (P_s*p = 0.08) was observed with HH having the highest gain:feed ratio (G:F) (0.74 ± 0.01), LH the lowest (0.70 ± 0.01) and the other two groups intermediate. In the growing-finishing phase, an interaction was observed between the piglet and sow diets with decreased G:F for LH (P_s*p = 0.04) and a tendency toward interaction with increased daily feed intake for LH (P_s*p = 0.07). The sow diet showed a tendency toward a long-lasting effect on the dressing percentage and meat thickness of the offspring, which was higher for the progeny of H sows (P_s < 0.01 and P_s = 0.02, respectively). At 23 weeks, serum urea concentrations tended to be lower for the HH and LL groups (P_s*p = 0.07). Fecal consistency scores were higher at day 10–day 14 after weaning for piglets from L sows (P_s = 0.03 and P_s < 0.01, respectively). At day 7 after weaning, fecal consistency score was higher for piglets fed the higher protein diet (P_p < 0.01). At 8 weeks of age, the apparent total tract digestibility of CP (ATTD_CP) interacted between piglet and sow diet (P_s*p = 0.02), with HH showing the highest digestibility values. In conclusion, the protein levels in sow late-gestation and piglet nursery diets interacted with feed efficiency, ATTD_CP and serum urea concentrations in the nursery phase.     

Nurse sow strategies in the domestic pig: I. Consequences for selected measures of sow welfare

Management strategies are needed to optimise the number of piglets weaned from hyper-prolific sows. Nurse sow strategies involve transferring supernumerary new-born piglets onto a sow whose own piglets are either weaned or fostered onto another sow. Such ‘nurse sows’ have extended lactations spent in farrowing crates, which could have negative implications for their welfare. This study used 47 sows, 20 of which farrowed large litters and had their biggest piglets fostered onto nurse sows which were either 1 week (2STEP7, n=9) or 3 weeks into lactation (1STEP21, n=10). Sows from which piglets were removed (R) were either left with the remainder of the litter intact (I) (remain intact (RI) sows, n=10), or had their litters equalised (E) for birth weight using piglets of the same age from non-experimental sows (remain equalised (RE) sows, n=9). Piglets from 2STEP7 were fostered onto another nurse sow which was 3 weeks into lactation (2STEP21, n=9). Back-fat thickness was measured at entry to the farrowing house, at fostering (nurse sows only) and weaning. Sows were scored for ease of locomotion and skin and claw lesions at entry to the farrowing house and weaning. Salivary cortisol samples were collected and tear staining was scored at 0900 h weekly from entry until weaning. Saliva samples were also taken at fostering. Data were analysed using GLMs with appropriate random and repeated factors, or non-parametric tests were applied where appropriate. Back-fat thickness decreased between entry and weaning for all sows (F_1,42=26.59, P<0.001) and tended to differ between treatments (F_4,16=2.91; P=0.06). At weaning RI sows had lower limb lesion scores than 2STEP7 and RE sows (χ²₄=10.8, P<0.05). No treatment effects were detected on salivary cortisol concentrations (P>0.05) and all nurse sows had a higher salivary cortisol concentration at fostering, compared with the other days (F_10,426=3.47; P<0.05). Acute effects of fostering differed between nurse sow treatments (F_2,113=3.45, P<0.05); 2STEP7 sows had a higher salivary cortisol concentration than 1STEP21 and 2STEP21 sows on the day of fostering. 2STEP7 sows had a higher salivary cortisol concentration at fostering, compared with 1STEP21 and 2STEP21 sows. Tear staining scores were not influenced by treatment (P>0.05). In conclusion, no difference was detected between nurse sows and non-nurse sows in body condition or severity of lesions. Although some nurse sows experienced stress at fostering, no long-term effect of the nurse sow strategies was detected on stress levels compared with sows that raised their own litter.     

Body composition and organ development of intra-uterine growth restricted pigs at weaning

Sow litter sizes have increased, subjecting more small piglets to intra-uterine growth restriction (IUGR). Research on the development and growth of IUGR pigs is limited. The objective of this study was to compare the body composition and organ development of IUGR pigs at weaning, and to estimate their growth performance from birth to 30 kg. A total of 142 IUGR and 142 normal piglets were classified at birth based on their head morphology. At weaning, 20 IUGR and 20 normal piglets were collected, a whole-body dual-energy x-ray absorption scan was performed, and the piglets were euthanized for organ measurements. Body weight (BW) was measured weekly from birth to 30 kg, rectal temperature and whole-blood glucose levels were measured weekly from birth to weaning, and blood samples were collected at days 7, 14 and 21 for IGF-1 analysis. Results showed that IUGR pigs have a similar percentage of adipose tissue (P > 0.05) compared to normal pigs at 24 days of age. Organs were smaller (P < 0.001) in IUGR pigs than in normal pigs, whereas brain, liver, lungs and adrenal glands were relatively larger (P < 0.05) in relation to the BW of IUGR pigs. Average birth weight (BiW) of normal pigs was greater (P < 0.001) compared with IUGR pigs (1.38 v. 0.75 kg), and the average daily gain (ADG) of IUGR pigs was reduced from day 0 to 14, day 0 to 28 (weaning) and from weaning to 30 kg compared to normal pigs. From birth to weaning at day 28, IUGR piglets had a 72.9 g/day greater fractional ADG (FADG) in relation to their BiW (P < 0.05), but FADG did not differ (P > 0.05) from weaning to 30 kg. Rectal temperature of IUGR piglets was greater (P < 0.05) on day 7 compared with normal piglets, and, even though blood glucose levels were decreased (P < 0.001) in IUGR piglets at day 0, neither glucose nor IGF-1 concentrations differed (P > 0.05) between IUGR and normal piglets. In conclusion, IUGR piglets exhibited some relatively larger organs at weaning compared to normal pigs, but body composition was similar between IUGR and normal pigs. In addition, IUGR pigs had a reduced ADG from birth to 30 kg, and, although they exhibited a greater FADG during nursing, IUGR pigs still require six additional days to reach a BW of 30 kg in comparison to normal pigs.     

The selection of trimethoprim resistant Escherichia coli in the faecal flora of piglets sucking a sow treated parenterally with trimethoprim

The parenteral administration of a therapeutic preparation containing trimethoprim to a lactating sow was associated with the appearance of trimethoprim resistant Escherichia coli in the faeces of the sow and her piglets. The E. coli isolates were differentiated into strains on the basis of their O-serogroup, biotype and resistance pattern. The majority (75%) of trimethoprim resistant strains were isolated only from the faeces of the piglets during the 14 d survey period. This suggested that the agent must have been excreted at a sufficiently high concentration in the milk to select for resistant strains present in the minority flora of the sucking pigs.     

Effect of a disinfection strategy on the methicillin‐resistant Staphylococcus aureus CC398 prevalence of sows,their piglets and the barn environment

Aims

To assess, in a cleaned and disinfected barn environment, the efficacy of an animal disinfection strategy to reduce the livestock‐associated methicillin‐resistant Staphylococcus aureus (LA‐MRSA) prevalence in sows, their offspring and the barn environment.

Methods and Results

On each farm, six sow rounds were sampled; sows were divided into either a test or control group. Per round, 20 sows and 40 of their piglets were sampled at different time points together with the barn environment. The disinfection strategy of the test groups consisted of washing the sows with a shampoo followed by disinfection of the skin with a solution containing chlorhexidine digluconate and isopropanol. On the first day of disinfection and 6 days after stopping the disinfection, a significant decrease (P < 0·01) of on average 68 and 66% in sow MRSA prevalence was observed on both farms, whereas no decrease was seen in the control groups. Just before weaning, 21–28 days after the end of the disinfection strategy, the difference in MRSA prevalence between both groups was reduced to 4% and no longer significant (P = 0·20). The MRSA prevalence of the piglets in the test groups was significantly lower (26%; P < 0·01) 6 days after the end of disinfection. Just before weaning, this difference was reduced to 5% but still significant (P < 0·01). In the swine nursery unit, no significant difference (P = 0·99) was seen between both groups. Based on semi‐quantitative counts, a relationship (r² > 0·6; P < 0·01) was seen between MRSA contamination in the barn environment and the MRSA prevalence in pigs.

Conclusion

Results show that the tested disinfection strategy reduces temporarily the sow and piglet MRSA status, but does not result in a final reduction in MRSA at weaning or in the nursery unit.

Significance and Impact of the Study

First report on the efficacy of an animal disinfection strategy to reduce LA‐MRSA prevalence in sows, their offspring and the barn environment.     

ExPEC-typical virulence-associated genes correlate with successful colonization by intestinal E. coli in a small piglet group

Upon studying the transmission of Escherichia coli from a sow to five of her piglets, we observed domination of the coliform flora in piglets by a single E. coli clone, especially after weaning. This haemolytic cloneH1 did not harbour any virulence determinants typical for intestinal pathogenic E. coli isolates from swine but had a virulence gene profile very similar to extraintestinal E. coli (ExPEC), including genes coding for P fimbriae and several iron acquisition systems, besides having an affiliation to the phylogenetic B2 group. Overall, we show that the presence of higher numbers of ExPEC-typical virulence-associated genes (VAGs) in clones correlate with their successful colonization ability in piglets. We conclude that VAGs typical for ExPEC also support intestinal colonization in healthy pigs. Faeces of healthy domestic pigs can harbour high numbers of ExPEC-similar E. coli and are suggested to be a potential risk for the transmission of such bacteria to other hosts.     

Effect of oral polyamine supplementation pre-weaning on piglet growth and intestinal characteristics

A high proportion of piglets fail to adapt to the changing composition of their diet at weaning, resulting in weight loss and increased susceptibility to pathogens. Polyamines are present in sow milk and promote neonatal maturation of the gut. We hypothesised that oral spermine and spermidine supplementation before weaning would increase piglet growth and promote gastrointestinal development at weaning. In Experiment One, one pair of liveweight (LW)-matched piglets per litter from first and third lactation sows received 2 ml of a 0 (Control) or 463 nmol/ml spermine solution at 14, 16, 18, 20 and 22 days of age (n=6 piglets/treatment per parity). Villus height and crypt depth in the duodenum and jejunum were measured at weaning (day 23 postpartum). In Experiment Two, piglets suckling 18 first and 18 third lactation sows were used. Within each litter, piglets received 2 ml of either water (Control), 463 nmol/ml spermine solution or 2013 nmol/ml spermidine solution at 14, 16, 18, 22 and 24 days of age (n=54 piglets/treatment per sow parity). Piglets were weighed individually at 14, 18, 24 (weaning) and 61 days of age. In Experiment One, oral spermine supplementation resulted in a 41% increase in villus height, a 21% decrease in crypt depth and 79% decrease in the villus height : crypt depth ratio compared with control piglets (P<0.01). In Experiment Two, spermine and spermidine-supplemented piglets suckling first lactation sows grew faster (P<0.05) between days 14 and 18 postpartum than control piglets: 0.230±0.011 and 0.227±0.012 v. 0.183±0.012 kg/day, respectively. Spermine supplementation tended (P<0.1) to increase piglet LW gain from weaning to day 37 post-weaning compared with control piglets (0.373±0.009 v. 0.341±0.010 kg/day). In conclusion, spermine supplementation increased villus height at weaning, and appears to have the potential to improve the pre- and post-weaning growth of conventionally weaned piglets.     

Effect of an Escherichia coli F4/F18 bivalent oral live vaccine on gut health and performance of healthy weaned pigs

Oral live vaccines stimulate host immunity, but they could also affect intestinal mucosa development and gut microbiota of piglets during the postweaning. The aim of this study was to determine the effect of an oral vaccine against Escherichia coli F4 and F18 (Coliprotec F4/F18®), on gut functionality and integrity, growth performance and health status of postweaning piglets. A total of 96 weaned piglets (23.30 ± 1.85 days of age; 7334 ± 1039 g BW) were divided into two groups (16 replicates/group; three piglets/replicate) as follows: (1) Control (CO), fed a standard diet (prestarter up to 14 days, then starter feed); (2) Treated (TRT): as CO but vaccinated with Coliprotec F4/F18® at weaning (day 0). Piglets were weighed at day 0 and weekly until day 35. Individual faecal score was recorded daily. Piglets were sacrificed at days 10 (1/3 of total) and 35 (2/3). Samples of jejunum mucosa and of cecum content were collected for morphometric, immunohistochemistry analysis and for microbiota profile analysis, respectively. Data were fitted using a linear model including treatment, class of starting BW as fixed factors and litter as random factor. From days 0 to 7, piglets from the TRT group tended to have a higher average daily gain (+22.6%, P = 0.08) and average daily feed intake compared to the CO group (+13.2%, P = 0.022). Gain to feed ratio was lower in the TRT group from days 14 to 35 (?6.6%, P = 0.011). From days 7 to 14, the TRT group had a higher diarrhoea index (?199%, P < 0.001). Crypt depth was higher in the CO group (+10.9%, P = 0.04) at day 10, but not at day 35. Jejunal expression of Claudin-4 (probability of having a score = 3) was higher in the TRT group at day 10 (CO = 1.50% vs TRT = 2.69%, P < 0.0001) and day 35 (CO = 1.29% vs TRT = 1.92%, P = 0.012). Oral vaccine affected beta diversity at day 10 (P = 0.040; R² = 0.05) and increased the abundance of specific taxa and genera in the cecum at day 10, including Prevotella (lg2FC = 23.2, FDR < 0.001). The results showed how an Escherichia coli-based vaccine supplied to weaned pigs can promote gut health by controlling symptoms of the postweaning perturbation in the first 2 weeks postweaning. In addition, the vaccine strains showed a probiotic-like effect by modulating gut microbiota favouring the establishment of beneficial bacteria, and by promoting gut barrier integrity.     

Effect of milk deprivation during the lactation period on performance and digestive enzyme activities of the piglets following weaning

Post-weaning performance of piglets from systems where lactation is disrupted (e.g. from multisuckling systems) is superior to conventionally reared piglets. The objective of this study was to establish whether restricted growth prior to weaning caused by disruption of suckling was an important factor in post-weaning performance and also whether there were related changes in gastro-intestinal development. Ten litters of eight piglets were used in a split-plot design. Half of each litter (limited suckling, LS) had suckling disrupted by separation from their dam for 7 h/day from day 14 to 28 after farrowing. The remainder of each litter was allowed to suck normally (normal suckling, NS). The same amount of creep feed was offered to LS piglets as consumed by NS littermates on the previous day. There were no differences in weight between LS and NS piglets at 14 days of age, but restricting access to the sow reduced weaning weight at 28 days of age (7.96 v. 9.00 kg; LS v. NS; P < 0.01; s.e.d. 0.23). Feed intakes were greater for LS than NS piglets over the first 28 days post weaning, particularly in the 1st week after weaning when feed efficiency was also improved (0.91 v. 0.62 kg gain per kg feed; P < 0.01; s.e.d. 0.08). As a result, LS piglets grew more rapidly in the first 28 days post weaning, particularly in the first 7 days after weaning. Subsequent performance to 8 weeks was similar for both groups. Digestive organ weights were not different at 2 and 9 days after weaning; nor were small intestine specific enzyme activities significantly different ( P>0.05). Pancreatic trypsin activity was, however, greater ( P < 0.01) for LS pigs on both days 2 and 9 post weaning. In conclusion the restriction of growth as a result of limited suckling itself is an important factor in determining post-weaning performance and may be related to development of pancreatic trypsin activity.     

Estimation of genetic trends from 1977 to 1998 of body composition and physiological state of Large White pigs at birth

Genetic trends for body composition and blood plasma parameters of newborn piglets were estimated through the comparison of two groups of pigs (G77 and G98, respectively) produced by inseminating Large White (LW) sows with semen from LW boars born either in 1977 or in 1998. Random samples of 18 G77 and 19 G98 newborn piglets were used for whole carcass and tissue sampling. Plasma concentrations of glucose, albumin and IGF-1 were determined on 75 G77 and 90 G98 piglets from 18 litters. The G98 piglets had less carcass dry matter, protein and energy (P < 0.01) than their G77 counterparts. When expressed in g/kg birth weight, livers were lighter (P < 0.001) and contained less glycogen (P < 0.01) in G98 piglets, with no difference in the activity of the hepatic glucose-6-phosphatase between G98 and G77 piglets. Concentrations of protein, DNA, RNA in longissimus dorsi muscle were unaffected by selection. Plasma concentrations of glucose (P < 0.05) and IGF-1 (P < 0.01) were lower in G98 than in G77 piglets. On the whole, the results suggest that the improvement in lean growth rate and in sow prolificacy from 1977 to 1998 has resulted in a lower maturity of piglets at birth.     

Group lactation from 7 or 14 days of age reduces piglet aggression at weaning compared to farrowing crate housing

Early life experiences can affect social behaviour in later life, but opportunities for socio-behavioural development are often overlooked in current husbandry practices. This experiment investigated the effects of rearing piglets in two-stage group lactation (GL) system from 7 or 14 days of age on piglet aggression at weaning. Three lactation housing treatments were applied to a total of 198 piglets from 30 litters of multiparous sows. All dams farrowed in standard farrowing crates (FCs). Group lactation litters were transferred with their dam at 7 (GL₇) or 14 days (GL₁₄) postpartum to GL pens (one pen of five sows at 8.4 m²/sow and one pen of seven sows at 8.1 m²/sow, per GL treatment). Farrowing crate litters remained with their dam in a single litter until weaning. At weaning, 10 to 14 piglets from two unfamiliar litters from the same housing treatment were mixed into pens (n=5 pens/treatment) and their behaviour was continuously recorded for 3.5 h. For each pen, the frequency of aggressive bouts (reciprocal and non-reciprocal aggression lasting <5 s), the frequency and duration of fights (reciprocal aggression lasting ⩾5 s) and bullying events (non-reciprocal aggression lasting ⩾5 s) were recorded, along with whether interactions involved familiar or unfamiliar piglets. Aggressive bouts delivered by FC piglets were approximately 1.5 and 3.0 times more frequent than that delivered by GL₇ and GL₁₄ piglets, respectively (40.5, 16.7 and 9.9 bouts/pig, respectively; P<0.05). Fighting was more frequent (1.6, 0.3 and 0.4 fights/pig, respectively; P<0.001) and fights were longer (83, 15 and 32 s fight/pig, respectively; P<0.001) between FC piglets than between GL₇ or GL₁₄ piglets. Bullying did not differ between housing treatments (P>0.05). GL₇ and GL₁₄ piglets engaged in a similar number of fights with unfamiliar as familiar piglets, but FC piglets had almost three times as many fights with unfamiliar than with familiar piglets (P<0.05). This experiment confirms the benefits of GL housing for pig social development. Further investigation is required to determine whether mixing before 14 days postpartum has implications for other indicators of animal welfare and productivity in a two-stage GL housing system.     

Response of performance characteristics and fecal consistency to long-lasting dietary supplementation with the probiotic strain Bacillus cereus var. toyoi to sows and piglets

Abstract

As part of an interdisciplinary research project, we studied the performance response of sows and their litters to the probiotic strain Bacillus cereus var. toyoi as well as feces consistency of piglets. Gestating sows (n = 26) were randomly allotted into two groups. The probiotic B. cereus var. toyoi was administered by dietary supplementation to one group of sows and their respective litters (probiotic group) whereas the second group (control group) received no probiotic supplementation. The duration of the application was nearly 17 weeks for sows (day 90 ante partum until day 28 post partum) and six weeks for piglets (day 15–56). Piglets were weaned after 28 days. Body weight and feed consumption were recorded weekly and fecal consistency of weaned piglets was studied daily. B. cereus var. toyoi was recovered from feces of sows and piglets as well as from digesta of piglets in the probiotic group, while being absent from all samples of control animals. In addition, the probiotic was detected in piglet feces and digesta before pre-starter feed was offered, indicating a second route of uptake besides diet. Sows of the probiotic group nursed numerically more piglets and supported a higher sum of total nursing days of all piglets within each litter than control sows (p = 0.04). In turn, body weight (BW) up to day 35 was greater for control piglets (p < 0.01), while average daily gain and gain to feed ratio (G:F) in weeks six and eight postweaning was higher in the probiotic group (p < 0.05). The overall G:F of the total postweaning period was 680 g/kg and 628 g/kg in the probiotic group and control group, respectively (p = 0.009). During the trial a high prevalence of liquid feces with its maximum in the second week after weaning was observed. Probiotic supplementation led to a reduction in the incidence of liquid feces and postweaning diarrhea by 38% and 59%, respectively (p < 0.001).     

Comparative Host Specificity of Human- and Pig- Associated Staphylococcus aureus Clonal Lineages

Bacterial adhesion is a crucial step in colonization of the skin. In this study, we investigated the differential adherence to human and pig corneocytes of six Staphylococcus aureus strains belonging to three human-associated [ST8 (CC8), ST22 (CC22) and ST36(CC30)] and two pig-associated [ST398 (CC398) and ST433(CC30)] clonal lineages, and their colonization potential in the pig host was assessed by in vivo competition experiments. Corneocytes were collected from 11 humans and 21 pigs using D-squame® adhesive discs, and bacterial adherence to corneocytes was quantified by a standardized light microscopy assay. A previously described porcine colonization model was used to assess the potential of the six strains to colonize the pig host. Three pregnant, S. aureus-free sows were inoculated intravaginally shortly before farrowing with different strain mixes [mix 1) human and porcine ST398; mix 2) human ST36 and porcine ST433; and mix 3) human ST8, ST22, ST36 and porcine ST398] and the ability of individual strains to colonize the nasal cavity of newborn piglets was evaluated for 28 days after birth by strain-specific antibiotic selective culture. In the corneocyte assay, the pig-associated ST433 strain and the human-associated ST22 and ST36 strains showed significantly greater adhesion to porcine and human corneocytes, respectively (p<0.0001). In contrast, ST8 and ST398 did not display preferential host binding patterns. In the in vivo competition experiment, ST8 was a better colonizer compared to ST22, ST36, and ST433 prevailed over ST36 in colonizing the newborn piglets. These results are partly in agreement with previous genetic and epidemiological studies indicating the host specificity of ST22, ST36 and ST433 and the broad-host range of ST398. However, our in vitro and in vivo experiments revealed an unexpected ability of ST8 to adhere to porcine corneocytes and persist in the nasal cavity of pigs.     

DNA Microarray-Based Identification of Serogroups and Virulence Gene Patterns of Escherichia coli Isolates Associated with Porcine Postweaning Diarrhea and Edema Disease

Escherichia coli strains causing postweaning diarrhea (PWD) and edema disease (ED) in pigs are limited to a number of serogroups, with O8, O45, O138, O139, O141, O147, O149, and O157 being the most commonly reported worldwide. In this study, a DNA microarray based on the O-antigen-specific genes of all 8 E. coli serogroups, as well as 11 genes encoding adhesion factors and exotoxins associated with PWD and ED, was developed for the identification of related serogroups and virulence gene patterns. The microarray method was tested against 186 E. coli and Shigella O-serogroup reference strains, 13 E. coli reference strains for virulence markers, 43 E. coli clinical isolates, and 12 strains of other bacterial species and shown to be highly specific with reproducible results. The detection sensitivity was 0.1 ng of genomic DNA or 10³ CFU per 0.3 g of porcine feces in mock samples. Seventeen porcine feces samples from local hoggeries were examined using the microarray, and the result for one sample was verified by the conventional serotyping methods. This microarray can be readily used to screen for the presence of PWD- and ED-associated E. coli in porcine feces samples.     

Kinetics of Transmission, Infectivity, and Genome Stability of Two Novel Mouse Norovirus Isolates in Breeding Mice

Murine noroviruses are a recently discovered group of viruses found within mouse research colonies in many animal facilities worldwide. In this study, we used 2 novel mouse norovirus (MNV) wildtype isolates to examine the kinetics of transmission and tissue distribution in breeding units of NOD.CB17-Prkdc^scid/J and backcrossed NOD.CB17-Prkdc^scid/J × NOD/ShiLtJ (N1) mice. Viral shedding in feces and dissemination to tissues of infected offspring mice were monitored by RT-PCR over a 6-wk period postpartum. Histologic sections of tissues from mice exposed to MNV were examined for lesions and their sera monitored for the presence of antibodies to MNV. Viruses shed in feces of parental and offspring mice were compared for sequence homology of the Orf2 gene. Studies showed that the wildtype viruses MNV5 and MNV6 behaved differently in terms of the kinetics of transmission and distribution to tissues of offspring mice. For MNV5, virus transmission from parents to offspring was not seen before 3 wk after birth, and neither isolate was transmitted between cages of infected and control mice. Susceptibility to infection was statistically different between the 2 mouse strains used in the study. Both immunodeficient NOD.CB17-Prkdc^scid/J mice and NOD.CB17-Prkdc^scid/J × NOD/ShiLtJ offspring capable of mounting an immune response shed virus in their feces throughout the 6-wk study period, but no gross or histologic lesions were present in infected tissues. Progeny viruses isolated from the feces of infected offspring showed numerous mutations in the Orf2 gene for MNV5 but not MNV6. These results confirm previous studies demonstrating that the biology of MNV in mice varies substantially with each virus isolate and mouse strain infected.Abbreviations: MNV, murine norovirus; MLN, mesenteric lymph nodes; NOD-scid, NOD.CB17-Prkdc^scid/J; VP1, viral protein 1The recent discovery of murine-specific noroviruses¹⁵ has stimulated concern in the laboratory animal health community regarding the potential for this group of viruses to cause disease in breeding colonies of mice or to negatively impact research with mice from norovirus infected colonies. Current knowledge of the biology of noroviruses in mice (MNV) is constrained by the limited number of virus isolates and mouse strains studied. One study¹⁵described the biologic and physicochemical properties of the original MNV1 isolated from mice deficient in a specific innate immune function. More recently, this innate immune deficiency has been mapped to STAT1 regulation of IFNαβ secretion.²¹Previous work¹⁵ demonstrated that inoculation of MNV1 into mouse strains deficient in the acquired immune response (129 RAG 2^−/−, B6 RAG1^−/−) resulted in the development of persistent infections with no evidence of disease, whereas inoculation of fully immunocompetent mice (129S6/SvEvTac) resulted in rapid elimination of MNV1, with viral RNA undetectable in the viscera by 3 d after inoculation. More recently, infections of outbred immunocompetent mouse strains with 3 wildtype isolates of MNV obtained from different geographic areas of the United States have been described.¹¹ Virus was detected in the feces and tissue of infected mice throughout the 8-wk study, suggesting that some isolates of MNV may persistently infect immunocompetent mice.The purpose of the present investigation was to extend the current knowledge of MNV by using 2 isolates of the virus in mouse strains that have not been previously used as infection models for MNV. We examined natural virus transmission from infected breeders to offspring, kinetics of infection within litters of infected breeding mice, and the pathogenesis of infection in breeding colonies of mice. In addition, we examined the effect of virus passage from parents to offspring on genomic stability of these 2 viral isolates. Exposure of offspring of immunodeficient mice and immunocompetent mice to the 2 different isolates of MNV resulted in different patterns of virus transmission, susceptibility to infection and kinetics of infection as shown by the progressive spread of virus within litters and in intestinal and extraintestinal tissues. MNV was shed persistently in the feces of all mice tested regardless of immune status, and viral progeny isolated from offspring mice contained genome sequence differences from the parent virus in the Orf2 gene, an area of the MNV genome known to be susceptible to mutations.     

Transfer of Lactic Acid Bacterial Strains from the Feed to the Sow,the Environment,and the Piglets

The spread of lactic acid bacterial strains to the environment and to newborn piglets was investigated after feeding of such strains to sows. Rifampicin resistant bacterial strains were fed to sows, 1010 c.f.u. per day, during the period from 1 week before expected farrowing until 1 week after farrowing. Fecal samples from the sows and samples of litter were collected for bacteriological examination together with swabs from the pens, the skin of the sows, and from the rectum of the piglets. The test strains were only excreted in relatively low amounts in the feces of the sows, approximately 103 −106 c.f.u. per gram. They were not able to displace the normal lactic acid bacterial flora in the sows nor were they transmitted to the intestinal tract of the piglets to any significant extent. After the last administration the test strains disappeared from both feces, skin, and environment, indicating that no permanent colonization had taken place, although considerable differences in duration of persistence were noticed between test strains.