Variability in lag-duration of Campylobacter spp. during enrichment after cold and oxidative stress and its impact on growth kinetics and reliable detection.

Campylobacter jejuni and Campylobacter coli continue to be the leading cause of zoonotic gastroenteritis in the European Union, making reliable detection in food important. Low storage temperatures and atmospheric oxygen concentrations during food production can cause sub-lethal damage or transient non-culturability which is why ISO 10272-1:2017 includes an enrichment step to repair cell damage and increase cell concentrations, thereby supporting detection of campylobacters from foods. The aim of this study was to assess the variability in lag-duration of C. jejuni and C. coli during enrichment after different food-relevant stress treatments and evaluate its impact on growth kinetics and reliability of detection outcomes. Therefore, 13 C. jejuni and 10 C. coli strains were subjected to cold stress during refrigerated and frozen storage. Refrigerated storage did not significantly reduce culturability, but frozen storage reduced cell concentrations by 1.6 ± 0.1 log10cfu/ml for both species. Subsequently, cells were enriched following ISO 10272-1:2017-A and cell concentrations were determined over time and lag-duration and growth rate were determined by fitting the Baranyi-model. Without prior stress treatment, mean lag-duration for C. jejuni and C. coli was 2.5 ± 0.2 h and 2.2 ± 0.3 h, respectively. Refrigerated storage increased lag-duration for C. jejuni to 4.6 ± 0.4 h and for C. coli to 5.0 ± 0.4 h and frozen storage increased lag-duration to 5.0 ± 0.3 h and 6.1 ± 0.4 h for C. jejuni and C. coli, respectively. Comparison of strain- and biological variability showed that differences in recovery after cold stress can be attributed mainly to strain variability since strain variability after refrigeration and freeze stress increased respectively 3-fold and 4-fold while biological variability remained constant. A subset of strains was also subjected to oxidative stress that reduced cell concentrations by 0.7 ± 0.2 log10 cfu/ml and comparison of recovery patterns after oxidative and freeze stress indicated that recovery behaviour was also dependent on the stress applied. A scenario analysis was conducted to evaluate the impact of heterogeneity in outgrowth kinetics of single cells on the reliability of detection outcomes following ISO protocol 10272-1:2017. This revealed that a 'worst-case'-scenario for successful detection by a combination of the longest lag-duration of 7.6 h and lowest growth rate of 0.47 h-1 still resulted in positive detection outcomes since the detection limit was reached within 32.5 h. This suggests that other factors such as competitive microbiota can act as a causative factor in false-negative outcomes of tested food samples.

Reducing viral contamination from finger pads: handwashing is more effective than alcohol-based hand disinfectants.

BACKGROUND: Hand hygiene is important for interrupting transmission of viruses through hands. Effectiveness of alcohol-based hand disinfectant has been shown for bacteria but their effectiveness in reducing transmission of viruses is ambiguous. AIM: To test efficacy of alcohol hand disinfectant against human enteric and respiratory viruses and to compare efficacy of an alcohol-based hand disinfectant and handwashing with soap and water against norovirus. METHODS: Efficacies of a propanol and an ethanol-based hand disinfectant against human enteric and respiratory viruses were tested in carrier tests. Efficacy of an alcohol-based hand disinfectant and handwashing with soap and water against noroviruses GI.4, GII.4, and MNV1 were tested using finger pad tests. FINDINGS: The alcohol-based hand disinfectant reduced the infectivity of rotavirus and influenza A virus completely within 30s whereas poliovirus Sabin 1, adenovirus type 5, parechovirus 1, and MNV1 infectivity were reduced <3 log10 within 3 min. MNV1 infectivity reduction by washing hands with soap and water for 30s (>3.0 ± 0.4 log10) was significantly higher than treating hands with alcohol (2.8 ± 1.5 log10). Washing with soap and water for 30s removed genomic copies of MNV1 (>5 log10), noroviruses GI.4 (>6 log10), and GII.4 (4 log10) completely from all finger pads. Treating hands with propanol-based hand disinfectant showed little or no reduction to complete reduction with mean genomic copy reduction of noroviruses GI.4, GII.4, and MNV1 being >2.6, >3.3, and >1.2 log10 polymerase chain reaction units respectively. CONCLUSIONS: Washing hands with soap and water is better than using alcohol-based hand disinfectants in removing noroviruses from hands.

Quantifying strain variability in modeling growth of Listeria monocytogenes.

Prediction of microbial growth kinetics can differ from the actual behavior of the target microorganisms. In the present study, the impact of strain variability on maximum specific growth rate (µmax) (h(-1)) was quantified using twenty Listeria monocytogenes strains. The µmax was determined as function of four different variables, namely pH, water activity (aw)/NaCl concentration [NaCl], undissociated lactic acid concentration ([HA]), and temperature (T). The strain variability was compared to biological and experimental variabilities to determine their importance. The experiment was done in duplicate at the same time to quantify experimental variability and reproduced at least twice on different experimental days to quantify biological (reproduction) variability. For all variables, experimental variability was clearly lower than biological variability and strain variability; and remarkably, biological variability was similar to strain variability. Strain variability in cardinal growth parameters, namely pHmin, [NaCl]max, [HA]max, and Tmin was further investigated by fitting secondary growth models to the µmax data, including a modified secondary pH model. The fitting results showed that L. monocytogenes had an average pHmin of 4.5 (5-95% prediction interval (PI) 4.4-4.7), [NaCl]max of 2.0mM (PI 1.8-2.1), [HA]max of 5.1mM (PI 4.2-5.9), and Tmin of -2.2°C (PI (-3.3)-(-1.1)). The strain variability in cardinal growth parameters was benchmarked to available literature data, showing that the effect of strain variability explained around 1/3 or less of the variability found in literature. The cardinal growth parameters and their prediction intervals were used as input to illustrate the effect of strain variability on the growth of L. monocytogenes in food products with various characteristics, resulting in 2-4 logCFU/ml(g) difference in growth prediction between the most and least robust strains, depending on the type of food product. This underlined the importance to obtain quantitative knowledge on variability factors to realistically predict the microbial growth kinetics.

Associations of conformation and locomotive characteristics in growing gilts with osteochondrosis at slaughter.

Osteochondrosis (OC) and abnormalities in conformation and locomotive characteristics (CLC) have been associated with premature culling in sows. Several CLC have been suggested to be associated with OC and might help as an in vivo indicator for and increased risk of having OC. The aim of this study was to investigate associations of OC with CLC assessed at several ages in growing gilts from 2 separate experiments over the effects of dietary restriction (Exp. 1) and floor type (Exp. 2) on OC prevalence. In Exp. 1, gilts (n = 211) were subjectively assessed for CLC at, on average, 4, 9, 11, 16, and 24 wk of age. In Exp. 2, gilts (n = 212) were subjectively assessed for CLC at, on average, 4, 9, 11, 16, and 22 wk of age. Assessment was done on 10 conformation and 2 locomotive characteristics using a 9-point grading scale by 2 observers. At, on average, 27 wk of age in Exp. 1 and 24 wk of age in Exp. 2, gilts were slaughtered and the knee, elbow, and hock joints were macroscopically assessed for OC. The CLC most frequently associated with OC were O shape or X shape of the hind legs, straight or bowed hind legs, and straight or sickled hock. X-shaped hind legs were associated with OC at slaughter in the knee joint at 4, 9, and 24 wk of age and at the animal level (all joints taken together) at 4, 9, and 16 wk of age. Straight or bowed hind legs were associated with OC at slaughter in the knee joint at 4 and 11 wk of age; in the hock joint at 11 wk of age; and at the animal level at 4, 9, 11, and 22 wk of age. Straight or sickled hock was associated with OC at slaughter in the knee joint at 4 wk of age, in the hock joint at 9 and 22 wk of age, and at the animal level at 9 and 22 wk of age. Results show that several CLC assessed at several ages were associated with OC, but consistent associations of a type of CLC in every assessment could not be found. The associations of CLC with OC are, therefore, difficult to be used as an in vivo indicator of increased risk for OC.

Quantifying variability on thermal resistance of Listeria monocytogenes.

Knowledge of the impact of strain variability and growth history on thermal resistance is needed to provide a realistic prediction and an adequate design of thermal treatments. In the present study, apart from quantifying strain variability on thermal resistance of Listeria monocytogenes, also biological variability and experimental variability were determined to prioritize their importance. Experimental variability was defined as the repeatability of parallel experimental replicates and biological variability was defined as the reproducibility of biologically independent reproductions. Furthermore, the effect of growth history was quantified. The thermal inactivation curves of 20 L. monocytogenes strains were fitted using the modified Weibull model, resulting in total 360 D-value estimates. The D-value ranged from 9 to 30 min at 55 °C; from 0.6 to 4 min at 60 °C; and from 0.08 to 0.6 min at 65 °C. The estimated z-values of all strains ranged from 4.4 to 5.7 °C. The strain variability was ten times higher than the experimental variability and four times higher than the biological variability. Furthermore, the effect of growth history on thermal resistance variability was not significantly different from that of strain variability and was mainly determined by the growth phase.

Inactivation of Listeria monocytogenes by disinfectants and bacteriophages in suspension and stainless steel carrier tests.

To simulate food contact surfaces with pits or cracks, stainless steel plates with grooves (depths between 0.2 and 5 mm) were constructed. These plates were artificially contaminated with Listeria monocytogenes in clean conditions, with organic soiling, or after 14 days of biofilm formation after which inactivation of the pathogen by Suma Tab D4 (sodium dichloroisocyanurate, 240 and 300 mg/liter), Suma Bac D10 (quaternary ammonium compound, 740 mg/liter), and bacteriophage suspension (Listex P100) was determined. Both chemical disinfectants performed well in suspension tests and in clean carrier tests according to the European standard with a reduction of more than 5 and 4 log units, respectively, of Listeria cells after 5 min of contact time. However, for the plates with grooves, the reduction could not meet the standard requirement, although a higher reduction of L. monocytogenes was observed in the shallow grooves compared with the deeper grooves. Furthermore, presence of food residues and biofilm reduced the effect of the disinfectants especially in the deep grooves, which was dependent on type of food substrate. Bacteriophages showed the best antimicrobial effect compared with the chemical disinfectants (sodium dichloroisocyanurate and quaternary ammonium compound) in most cases in the shallow grooves, but not in the deep grooves. The chlorine based disinfectants were usually less effective than quaternary ammonium compound. The results clearly demonstrate that surfaces with grooves influenced the antimicrobial effect of the chemical disinfectants and bacteriophages because the pathogen is protected in the deep grooves. The use of bacteriophages to inactivate pathogens on surfaces could be helpful in limited cases; however, use of large quantities in practice may be costly and phage-resistant strains may develop.

The influence of floor type before and after 10 weeks of age on osteochondrosis in growing gilts.

Osteochondrosis (OC) is a degenerative joint condition developing in a short time frame in young growing gilts that may cause lameness at an older age, affecting welfare and leading to premature culling of breeding sows. Causes of OC are multifactorial including both genetic and environmental factors. Floor type has been suggested to affect OC prevalence and effects might be age dependent during the rearing period. The aim of this study was to investigate possible age-dependent effects of floor type, conventional concrete partially slatted versus wood shavings as deep bedding, on OC prevalence in gilts (Dutch Large White × Dutch Landrace) at slaughter (24 wk of age; 106.5 [14.7 SD] kg of BW). At weaning (4 wk of age; 6.9 [1.3 SD] kg of BW), 212 gilts were subjected to 1 of 4 flooring regimens. Gilts were either subjected to a conventional floor from weaning until slaughter (CC), wood shavings as bedding from weaning until slaughter (WW), a conventional floor from weaning until 10 wk of age after which gilts were switched to wood shavings as bedding (CW), or wood shavings as bedding from weaning until 10 wk of age after which gilts were switched to a conventional floor (WC). After slaughter the elbow, hock, and knee joints were macroscopically examined for OC and scored on a 5 point scale where 0 indicates no OC and 4 indicates the severest form of OC. There was no significant difference (P > 0.4) between treatments on the overall OC prevalence for any joint assessed or at the animal level (all joints combined). At the animal level, however, gilts had greater odds to have OC scores 3 and 4 in the CW treatment (odds ratios [OR] = 2.3; P = 0.05), WC treatment (OR = 2.6; P = 0.02), and WW treatment (OR = 3.7; P < 0.001) compared with gilts in the CC treatment. The results indicate that there are no age-dependent effects of floor types on overall OC prevalence. However, wood shavings as bedding seems to increase the odds for severe OC and might affect animal welfare in the long term.

Cystic ovaries in intermittently-suckled sows: follicle growth and endocrine profiles.

This paper presents follicle development and hormone profiles for sows with normal ovulation or cystic follicles during an intermittent-suckling (IS) regime that started at Day 14 of lactation. Sows were subjected to separation from their piglets during blocks of 6h or 12h. In total, 8 out of 52 sows developed cystic follicles; either full cystic ovaries (n=6) or partial ovulation (n=2). Increase in follicle size of these sows was similar to that of normal ovulating sows until pre-ovulatory size at Day 5 after the start of separation, but from then on became larger (P<0.05). LH surge was smaller or absent in sows that developed (partially) cystic ovaries (0.4 ± 0.1 vs 3.6 ± 0.3 ngmL(-1); P<0.01). Peak levels of oestradiol (E2) were similar but high E2 levels persisted in sows that developed (partly) cystic ovaries and duration of oestrus tended to be longer. The risk of developing (partly) cystic ovaries was higher when IS occurred in blocks of 6h versus 12h (33 vs 10%). In conclusion, the appearance of cystic ovaries at approximately Day 20 of ongoing lactation was related to an insufficient LH surge, as is also the case in non-lactating sows.

The influence of dietary restriction before and after 10 weeks of age on osteochondrosis in growing gilts.

Osteochondrosis (OC) is one of the main causes of leg weakness causing premature culling in breeding sows and develops in a short time frame in young growing gilts. Dietary restriction may have different effects on OC prevalence depending on the age of the gilts. The aim of this study is to investigate age-dependent effects of dietary restriction, ad libitum vs. restricted (80% of ad libitum), on the occurrence of OC in gilts at slaughter (26 wk of age). At weaning (4 wk of age), 211 gilts were subjected to one of 4 treatments of a feeding regime. Gilts were administered either ad libitum feeding from weaning until slaughter (AA); restricted feeding from weaning until slaughter (RR); ad libitum feeding from weaning until 10 wk of age, after which gilts were switched to restricted feeding (AR); or restricted feeding from weaning until 10 wk of age, after which gilts were switched to ad libitum feeding (RA). At slaughter, the elbow, hock, and knee joints were harvested. Joints were scored macroscopically for articular surface deformations indicative of OC. Gilts in the RA treatment had significantly higher odds of being affected by OC than gilts in the RR and AR treatments in the hock joint (OR=3.3, P=0.04 and OR=8.5, P=0.002, respectively) and at animal level (OR=2.5, P=0.001 and OR=1.9, P=0.01, respectively). Gilts in the AA treatment had higher odds of being affected by OC than gilts in the AR treatment in the hock joint (OR=5.3, P=0.01). The results indicate a possible pathway to reduce the prevalence of OC in breeding gilts that will have to last several parities. Switching from restricted feeding to ad libitum feeding after 10 wk of age increases OC prevalence as opposed to restricted feeding after 10 wk of age.

Associations between osteochondrosis and conformation and locomotive characteristics in pigs.

Conformation and locomotive characteristics (CLC; i.e., leg conformation and gait movement patterns) may be associated with osteochondrosis (OC) in pigs. Osteochondrosis and CLC increase the risk of premature culling. This study investigated whether CLC have an explanatory value, over the previously modeled effects of sex, feeding, and housing conditions, on the occurrence and severity of OC in several joints and at the animal level. At 154 to 156 d of age, 267 pigs were subjectively scored on 9 conformation and 2 locomotive characteristics. Scoring was performed on a 9-point linear grading scale. For conformation characteristics, score 5 indicated normal conformation and scores 1 and 9 indicated severe deviations from normal. For the locomotive characteristics, score 1 indicated normal locomotion and score 9 indicated severe deviation from normal. At 161 to 176 d of age, pigs were slaughtered and joints were dissected for macroscopic evaluation of OC status. Results showed that swaying hindquarters and a stiffer gait were associated with greater scores for OC in, respectively, the femoropatellar (P = 0.018) and tarsocrural joint (P = 0.005), smaller inner claws as compared with the outer claws of the front legs was associated with lower scores for OC than equally sized claws in the femoropatellar joint (P = 0.021) and on animal level (P = 0.010), steep and weak pasterns of the front legs were associated with greater scores for OC in the elbow joint (P = 0.004) and on animal level (P = 0.018), X-shaped hind legs was associated with greater scores for OC on animal level (P = 0.037), and steep and weak pasterns of the hind legs were associated with lower scores for OC than normal conformation in the tarsocrural joint (P = 0.05). This study found several CLC that were associated with OC in several joints and at an animal level. This study showed that certain CLC might be used as indicators of OC and included in the criteria for selection of replacement animals for the breeding herd.

Progestagen supplementation during early pregnancy does not improve embryo survival in pigs.

Progesterone supplementation during early pregnancy may increase embryo survival in pigs. The current study evaluated whether oral supplementation with an analogue of progesterone, altrenogest (ALT), affects embryo survival. A first experiment evaluated the effect of a daily 20-mg dosage of ALT during days 1-4 or 2-4 after onset of oestrus on embryo survival at day 42 of pregnancy. A control group (CTR1) was not treated. The time of ovulation was estimated by transrectal ultrasound at 12-h intervals. Altrenogest treatment significantly reduced pregnancy rate when start of treatment was before or at ovulation: 25% (5/20) compared to later start of treatment [85% (28/33)] and non-treated CTR1 [100% (23/23)]. Altrenogest treatment also reduced (p < 0.05) number of foetuses, from 14.6 ± 2.6 in CTR1 to 12.5 ± 2.5 when ALT started 1-1.5 days from ovulation and 10.7 ± 2.9 when ALT started 0-0.5 days from ovulation. In a second experiment, sows with a weaning-to-oestrous interval (WOI) of 6, 7 or 8-14 days were given ALT [either 20 mg (ALT20; n = 49) or 10 mg (ALT10; n = 48)] at day 4 and day 6 after onset of oestrus or were not treated (CTR2; n = 49), and farrowing rate and litter size were evaluated. Weaning-to-oestrous interval did not affect farrowing rate or litter size. ALT did not affect farrowing rate (86% vs 90% in CTR2), but ALT20 tended to have a lower litter size compared with CTR2 (11.7 ± 4.1 vs 13.3 ± 3.1; p = 0.07) and ALT10 was intermediate (12.3 ± 2.9). In conclusion, altrenogest supplementation too soon after ovulation reduces fertilization rate and embryo survival rate and altrenogest supplementation at 4-6 days of pregnancy reduces litter size. As a consequence, altrenogest supplementation during early pregnancy may reduce both farrowing rate and litter size and cannot be applied at this stage in practice as a remedy against low litter size.

Antibacterial activity of reactive quaternary ammonium compounds in solution and in nonleachable coatings.

Antibacterial polymers suitable for coating applications without leaching of the biocidal component have been obtained by UV copolymerization of acrylic resins with acrylic monomers containing quaternary ammonium moieties. Suitable reactive biocides, based on quaternary ammonium monomers (QAMs), endowed with undecylacryloyl group and alkyl chains with 2 (QAM-C2), 8 (QAM-C8), and 16 (QAM-C16) carbon atoms have been synthesized. Aqueous solutions of QAMs showed biocidal activity against Escherichia coli, Staphylococcus aureus, and Listeria monocytogenes strains both in suspension and adhered to stainless steel surfaces. QAM-C16 and QAM-C8 evidenced higher activity toward bacteria in suspension and on stainless steel, respectively. The QAMs have shown sufficient reactivity to be copolymerized, by UV irradiation, with a commercial urethane acrylic resin for coating. Bioactivity tests, performed on free films of crosslinked coatings containing 1% of copolymerized QAM, have shown an increasing inactivation effect in the order of magnitude L. monocytogenes, E. coli, S. aureus with a maximum activity of the QAM-C8.

Survival of Listeria monocytogenes on a conveyor belt material with or without antimicrobial additives.

Survival of Listeria monocytogenes on a conveyor belt material with or without antimicrobial additives, in the absence or presence of food debris from meat, fish and vegetables and at temperatures of 10, 25 and 37 degrees C was investigated. The pathogen survived best at 10 degrees C, and better at 25 degrees C than at 37 degrees C on both conveyor belt materials. The reduction in the numbers of the pathogen on belt material with antimicrobial additives in the first 6h at 10 degrees C was 0.6 log unit, which was significantly higher (P<0.05) than the reduction of 0.2 log unit on belt material without additives. Reductions were significantly less (P<0.05) in the presence of food residue. At 37 degrees C and 20% relative humidity, large decreases in the numbers of the pathogen on both conveyor belt materials during the first 6h were observed. Under these conditions, there was no obvious effect of the antimicrobial substances. However, at 25 degrees C and 10 degrees C and high humidity (60-75% rh), a rapid decrease in bacterial numbers on the belt material with antimicrobial substances was observed. Apparently the reduction in numbers of L. monocytogenes on belt material with antimicrobial additives was greater than on belt material without additives only when the surfaces were wet. Moreover, the presence of food debris neutralized the effect of the antimicrobials. The results suggest that the antimicrobial additives in conveyor belt material could help to reduce numbers of microorganisms on belts at low temperatures when food residues are absent and belts are not rapidly dried.

Effect of excessive, hormonally induced intrauterine crowding in the gilt on fetal development on day 40 of pregnancy.

Selection for litter size may result in an increase in uterine crowding due to a faster increase in ovulation rate than in litter size. Increased ovulation rate does not result in a proportionally increased number of piglets born alive. In this study, the effect of ovulation rate on vitality characteristics of fetal-placental units at d 40 of pregnancy was investigated. For this, 43 Large White gilts were treated with hormones to induce superovulation. Average ovulation rate was 45.16 +/- 13.22; average number of vital fetuses at d 40 of pregnancy was 17.09 +/- 3.61 that weighed 11.26 +/- 1.99 g; their placenta weighed 31.88 +/- 14.79 g; and they occupied 11.69 +/- 4.90 cm of the uterus. Loss in oocytes (i.e., that did not result in a vital fetus at d 40) increased with increasing ovulation rate and occurred before (early mortality; P = 0.0003) and after implantation (late mortality, i.e., traces visible at d 40; P < 0.0001). With respect to the vital fetuses, increased ovulation rate resulted in decreased fetal (P = 0.0008) and placental weight (P = 0.0008) and decreased length of the area in the uterus that was occupied by the placenta (P = 0.0011). Strong correlations existed between placental and fetal weight [0.68; 95% confidence interval (CI) = 0.64 to 0.72], and placental weight and length (0.78; 95% CI = 0.74 to 0.82). Fetal-placental characteristics were weakly correlated to distance to the implantation sites of neighboring fetuses, a measure of crowdedness [-0.002 (95% CI = -0.042 to 0.038) with fetal weight to 0.16 (95% CI = 0.12 to 0.20) with placental length]. Increased ovulation rates, but more specifically increased late mortality rates, have negative effects on the remaining vital fetuses with respect to the fetal (P = 0.0085) and placental weight (P < 0.0001) and length of the implantation site (P = 0.0016). The most extreme effect was on placental weight, in which a uterus with <10 cases of late mortality was on average 25% greater than placental weight in a uterus with >18 cases of late mortality (P < 0.0001). Furthermore, increased ovulation rates resulted in decreased within litter variation for fetal (P = 0.0018) and placental weight (P = 0.0084). At increased ovulation rates, the number of live fetuses remained similar, but placental development is impaired and the growth of the fetus is retarded compared with reduced ovulation rate, with effects likely lasting into adult life.

Ovarian responses to lactation management strategies.

A number of lactation management strategies can be applied to reduce negative effects of lactation on post-weaning fertility. This paper focuses on effects of lactation length, Intermittent Suckling and Split Weaning on follicle development and subsequent oestrus. It is concluded that a lactation length of less than 3 weeks still leads to suboptimal reproductive performance in our modern sows. Further, both Intermittent Suckling and Split Weaning stimulate lactational follicle development and oestrus, but the variation in response between sows still limits practical application.

LH and FSH secretion, follicle development and oestradiol in sows ovulating or failing to ovulate in an intermittent suckling regimen.

The present paper describes LH and FSH secretion, follicle development and ovulation in sows that were subjected to a limited nursing regimen. From Day 14 of lactation, 32 sows were separated from their piglets for 12 h every day (intermittent suckling; IS). Half the sows had boar contact during separation. Nine of 32 sows ovulated spontaneously within 14 days from initiation of IS. The frequency of LH pulses on the first day of IS tended to be higher in anovulatory sows (6.3 v. 4.2 pulses per 12 h; P < 0.10); other characteristics of LH secretion were similar to sows that ovulated. The characteristics of FSH secretion did not differ over the 8-h sampling period. Boar contact did not influence either LH and FSH secretion or the number of sows that ovulated. Up to 58% of anovulatory sows showed an increase in follicle size after initiation of IS and, 4 days after the initiation of IS, one-third still had follicles similar in size to those in ovulatory sows. However, the oestradiol concentration in anovulatory sows did not increase. We conclude that FSH and LH stimulation in anovulatory sows is not limiting for normal follicle development, but that ovarian follicles are not responsive to increased LH secretion.

Campylobacter jejuni: a study on environmental conditions affecting culturability and in vitro adhesion/invasion.

AIMS: Nongrowing cultures of Campylobacter jejuni lose their culturability. It is unclear whether this loss in culturability also affects their ability to interact with host cells. The purpose of this study was to determine the relevance of the number of culturable cells to the ability to adhere/invade in Caco-2 cells. METHODS AND RESULTS: For C. jejuni C356, culturability and adhesion/invasion were monitored in time (days) under different storage conditions (temperature, medium, atmosphere). Decrease rates of both culturability and adhesion/invasion were dependent on the conditions used, but the number of adhering/invading cells per culturable cell was not affected by the environmental conditions. Furthermore five strains were monitored at one condition. The culturability and adhesion/invasion decrease rates did not significantly differ per strain; however the number of adhering/invading cells per culturable cell was strain dependent. CONCLUSIONS: Culturability and adhesion/invasion of C. jejuni are linearly related. The number of adhering/invading cells per culturable C. jejuni cell is strain dependent, but is not affected by environmental conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: It was shown that the number of culturable cells is a good measure for the in vitro adhesion/invasion of. C. jejuni.

Intermittent suckling enables estrus and pregnancy during lactation in sows: effects of stage of lactation and lactation during early pregnancy.

Previously we demonstrated that pre-ovulatory LH and post-ovulatory progesterone (P4) concentrations in plasma were low and embryo development was retarded when sows were induced to ovulate during lactation by submitting them to intermittent suckling (IS). The present study investigated whether this was due to: (1) stage of lactation when IS was initiated, and (2) continuation of IS post-ovulation. Multiparous Topigs40 sows were studied under three conditions: conventional weaning at Day 21 of lactation (C21; n=30), intermittent suckling from Day 14 of lactation (IS14; n=32), and intermittent suckling from Day 21 of lactation (IS21; n=33). Sows were separated from piglets for 12h daily during IS. IS sows were either weaned at ovulation or 20d following ovulation. One-third (21/63) of the IS21 and C21 sows had already ovulated or had large pre-ovulatory follicles at Day 21 and were excluded from further study. Initiation of IS at Day 14 instead of Day 21 of lactation tended to reduce P4 at 7d post-ovulation (P=0.07), did not affect pregnancy rate, and tended to reduce embryo survival (P=0.06). Continuation of IS during pregnancy resulted in lower P4 at 7 and 12d post-ovulation, tended to reduce embryo weight and pregnancy rate (P<0.10), whereas embryo survival was not affected. This study presents data for a population of sows in which follicle growth and ovulation are easily triggered under suckling conditions. Further, when these sows are bred during lactation, initiation of IS at 21 rather than 14d of lactation with weaning at ovulation yields the most desirable reproductive performance.

The intermittent suckling regimen in pigs: consequences for reproductive performance of sows.

Intermittent suckling (IS), a system in which the sow and the piglets are separated for a number of hours per day during lactation, is put forward as a system that can increase piglet welfare around weaning and also induce lactational oestrus. To be of practical use, IS regimes need to result in a predictable oestrus and good reproductive performance. This review describes the effects of IS on sow reproductive performance. During IS, the LH pulsatility pattern switches to a high frequency, low amplitude release during separation, as is normally observed at weaning. In sows that ovulate during IS, LH pulsatility remains higher than in anovulatory sows. Around 90% of the IS sows show follicle growth up to pre-ovulatory size in a similar time period as sows post-weaning. Depending on the stage of lactation that IS commenced, 13% (day 21) to 21% (day 14) of the sows with pre-ovulatory follicles did not ovulate, but follicles either regressed or developed into cystic follicles. In the ovulatory IS sows oestradiol production is comparable but the LH surge is lower with an early start of IS (day 14) when compared with the weaned sows. Continuation of IS post-ovulation has negative effects on pregnancy rate, embryo development and progesterone levels. Factors related to lactation (e.g. a high metabolic clearance rate), may play a role in this. In conclusion, IS can result in a fertile oestrus when the timing of start of IS and duration of IS are taken into account, but for practical use IS regimes result in a too variable reproductive performance.

Blastocyst morphology, actin cytoskeleton quality and chromosome content are correlated with embryo quality in the pig.

Embryo survival rates obtained after transfer of in vitro produced porcine blastocysts are very poor. This is probably related to poor quality of the embryos. The aim of the present study was to determine markers for good quality blastocysts. Therefore, we tried to link blastocyst morphology to several morphological and cell biological properties, and evaluated the survival of in vitro produced, morphologically classified, blastocysts following non-surgical transfer. In vitro and in vivo produced blastocysts were allocated to two groups (classes A and B) on the basis of morphological characteristics. The quality of their actin cytoskeleton, their total cell number, their ability to re-expand after cytochalasin-B treatment and the occurrence of numerical chromosome aberrations were studied and compared. In vivo produced blastocysts were used as a control. Our results indicate that the ability of blastocysts to re-expand after cytochalasin-B-induced actin depolymerization was positively correlated with the morphology of the blastocyst, and associated with the quality of the actin cytoskeleton. Chromosome analysis revealed that mosaicism is inherent to the in vitro production of porcine embryos, but also that in vivo produced blastocysts contained some non-diploid cells. In non-surgical embryo transfer experiments more recipients receiving class A blastocysts were pregnant on Day 20 than those receiving class B blastocysts. One recipient gave birth to six piglets from class A in vitro produced blastocysts, providing a verification of the enhanced viability of blastocysts that were scored as 'good' on the basis of their morphology.