Method for collecting and immobilizing individual cumulus cells enabling quantitative immunofluorescence analysis of proteins.

Most immunofluorescence methods rely on techniques dealing with a very large number of cells. However, when the number of cells in a sample is low (e.g., when cumulus cells must be analyzed from individual cumulus-oocyte complexes), specific techniques are required to conserve, fix, and analyze cells individually. We established and validated a simple and effective method for collecting and immobilizing low numbers of cumulus cells that enables easy and quick quantitative immunofluorescence analysis of proteins from individual cells. To illustrate this technique, we stained proprotein of a disintegrin and metalloproteinase with thrombospondin-like repeats-1 (proADAMTS-1) and analyzed its levels in individual porcine cumulus cells.

Development and validation of a new TD-GC/MS method and its applicability in the search for human and animal decomposition products.

Differentiation between human and animal remains by means of analysis of volatile compounds released during decomposition is impossible since no volatile marker(s) specific for human decomposition has been established today. Hence, the identification of such a marker for human decomposition would represent great progression for the discovery of buried cadavers by analytical techniques. Cadaver dogs can be trained more efficiently, the understanding of forensic entomology can be enhanced, and the development of a portable detection device may be within reach. This study describes the development and validation of a new analytical method that can be applied in the search of such (a) specific marker(s). Sampling of the volatile compounds released by decomposing animal and human remains was performed both in a laboratory environment and outdoors by adsorption on sorbent tubes. Different coatings and several sampling parameters were investigated. Next, the volatile compounds were analyzed and identified by a thermal desorber combined with gas chromatography coupled to mass spectrometry (TD-GC/MS). Different GC columns were tested. Finally, the analytical method was validated using a standard mixture of nine representative compounds.

Current knowledge on the transport and fate of spermatozoa in the reproductive tract of the bitch.

Canine sperm transport, distribution, storage and detachment is a complex, dynamic and highly regulated process. Transport of sperm within the bitch's reproductive tract is rapid and is influenced by the method of semen deposition (natural mating or artificial insemination) and by the timing of breeding in relation to the day of ovulation. The fertile lifespan of spermatozoa in the reproductive tract of the bitch is considerably longer than in most other domestic species, and the main sperm reservoirs appear to be the uterine crypts and the distal part of the uterotubal junction, where spermatozoa attach by their heads to uterine epithelium. While several in vitro studies demonstrated prolonged motility and viability of canine spermatozoa after coincubation with uterine tube explants, spermatozoal storage has not been documented in the canine uterine tube isthmus or ampulla in vivo. Several factors, including exposure to progesterone, solubilized zona pellucida proteins and post-ovulation uterine tube fluid, appear to trigger membrane events resulting in capacitation-like changes with subsequent motility pattern changes (transitional and hyperactivated) that are associated with sperm detachment. After mating or insemination, a normal low-magnitude post-mating uterine inflammatory response occurs, evidenced by an influx of polymorphonuclear neutrophils (PMNs), increased uterine contractions and an increased uterine artery blood flow. Recently, it was also shown that normal dogs with cystic endometrial hyperplasia develop a more significant endometritis, show fewer mating-induced uterine contractions, a decreased ability of spermatozoa to bind to uterine explants in vitro and a slower uterine clearance after mating.

Evaluation of a heating protocol and stocking density impact on heatstressed fattening pigs.

As climate change intensifies, heat stress mitigation for pigs becomes more important. Trials involving induced heat waves are useful to test several measures (e.g. reduced stocking density) at a faster rate, but only when accurately evaluated and validated. In the present study, we investigated the suitability of an artificial heating protocol at different pig weights (experiment 1). The impact of different stocking densities on fattening pigs during an artificial heat wave (experiment 2) was also investigated. Experiment 1: Forty 20-week-old pigs weighing 96.5 ± 7.3 kg (W100) and forty 17-week-old pigs weighing 72.7 ± 9.9 kg (W70) were housed in two compartments. An artificial heat wave (heat load) was induced for 3 days. During 3-day periods before, during and after the heat load, physiological parameters (respiration rate (RR), rectal temperature (Trectal), skin temperature (Tskin) and behavior) were measured and average daily feed intake was observed. Ambient temperature, relative humidity and temperature-humidity index (THI) were monitored. Experiment 2: A total of 150 fattening pigs were randomly divided into three treatment groups: SD1.3 (1.3 m2/pig), SD1.0 (1.0 m2/pig) and SD0.8 (0.8 m2/pig). All pens had a total pen surface of 4.88 m2, corresponding with 4, 5 and 6 fattening pigs in the SD1.3, SD1.0 and SD0.8 groups, respectively. The heat load was induced for 7 days on week 21. Respiration rate and Trectal were observed as in experiment 1. Average daily gain and average daily feed intake were also noted. During the heat load, THI reached ≥ 75 (78.4 (experiment 1) and 78.6 (experiment 2)), even when relative humidity decreased to ± 45%. Every physiological parameter showed significant increases during the heat load. The prolonged heating protocol in experiment 2 also provoked significant decreases in average daily feed intake (15%) and average daily gain (19%) for all groups. Weight within the studied range of 70-100 kg did not have a significant impact on any of the parameters. However, Tskin was affected by both weight and heat load (P < 0.05), where Tskin from W100 was always lower in comparison to W70. In addition, we found that 0.8 m2/pig doubled the increase of Trectal during the heat load, namely SD0.8 (0.22 °C) compared to SD1.0 (0.12 °C) (P = 0.033) and SD1.3 (0.13 °C) (P = 0.053). This suggests that pigs housed at higher densities are less able to regulate their internal heat production. However, RR and performances were not significantly affected by heat load in this experimental set-up. A stocking density of 1.0 m2/animal may be sufficient to mitigate some negative effects of heat stress.

Management factors associated with sow reproductive performance after weaning.

To achieve optimal reproductive performance in pig herds, sows need to become pregnant as soon as possible after weaning. The aim of this study was to investigate herd and management factors associated with reproductive performance of sows after weaning. A questionnaire pertaining to sow management at weaning and herd reproductive data were collected from 76 randomly selected commercial pig herds in Belgium. Associations between the herd factors and two reproductive parameters after weaning (weaning-to-oestrous interval: WEI and percentage of repeat breeders: RB) were analysed using general linear mixed models. A separated feeding strategy of breeding gilts from 60 kg onwards was significantly associated with a shorter WEI (5.54 vs 7.28 days; p = 0.040). Factors significantly associated with a lower percentage of RB were housing the newly weaned sows separated from the gestating sows (7% vs 12%; p = 0.003), using semen < 4 days after collection (7-9 vs 14%; p = 0.014) and stimulating oestrus twice a day (8 vs 11%; p = 0.025). In conclusion, some management practices, such as feeding strategy of breeding gilts, housing conditions of sows, method of oestrous stimulation and storage duration of semen, have an influence on the outcome of reproductive parameters such as weaning-to-oestrous interval and percentage of repeat breeders. These practices can be implemented rather easily by pig producers and may consequently lead to improvements of reproductive performance of sows after weaning.

Inhibitors of zinc-dependent metalloproteases hinder sperm passage through the cumulus oophorus during porcine fertilization in vitro.

In this study, we report for the first time on a possible contribution of metalloproteases in sperm passage through the cumulus matrix in pigs. The presence of 20 µM 1,10-phenanthroline (1,10-PHEN), inhibitor of zinc-dependent metalloproteases, strongly inhibited the degree of sperm penetration in cumulus-intact (CI), but not in cumulus-free (CF), porcine oocytes during IVF. The inhibitory effect of 1,10-PHEN was due to the chelation of metal ions as a non-chelating analog (1,7-PHEN) did not affect IVF rates. Furthermore, incubation with 1,10-PHEN did not affect sperm binding to the zona pellucida nor sperm motility, membrane integrity, or acrosomal status. These findings led to the assumption that 1,10-PHEN interacts with a sperm- or cumulus-derived metalloprotease. Metalloproteases are key players in physiological processes involving degradation or remodeling of extracellular matrix. In vivo, their proteolytic activity is regulated by tissue inhibitors of metalloproteases (TIMP1-TIMP4). We tested the effect of TIMP3 on fertilization parameters after porcine IVF. Similar to 1,10-PHEN, TIMP3 inhibited total fertilization rate of CI but not CF oocytes and did not influence sperm quality parameters. Although the inhibitory effect was stronger in CI oocytes, TIMP3 also reduced the degree of sperm penetration in CF oocytes, suggesting the involvement of a metalloprotease in a subsequent step during fertilization. In conclusion, our results indicate the involvement of TIMP3-sensitive, zinc-dependent metalloprotease activity in sperm passage through the cumulus oophorus in pigs. The results should provide the basis for further biochemical research toward the localization and identification of the metalloprotease involved.

Computer-assisted sperm analysis in dogs and cats: an update after 20 years.

In dogs and cats, computer-assisted sperm analysis (CASA) was originally described almost 20 years ago. Subsequently, numerous CASA systems were validated and used for various applications in dogs and to a lesser extent in cats. CASA systems offer an accurate, rapid, objective and simultaneous assessment of different semen parameters allowing the visualization of subtle changes in sperm characteristics, which cannot be identified by conventional semen analysis. The main problems of these computerized measuring devices are the relatively high investment costs and the need for standardization and validation before any practical use is possible. In comparison with automated motility and concentration assessment, automated morphometry and morphology assessment is more complex and time-consuming. Once validated, CASA systems can be routinely used in veterinary centres for assessment of fertility and for the improvement of sperm diluters, cooling and cryopreservation procedures in dogs and cats. Furthermore, information obtained by CASA systems could also be important when monitoring for example the effect of environmental stress on spermatozoa and for toxicity studies. In cats, CASA is less documented, and most studies describe the characteristics of epididymal sperm, which is frequently used for in vitro fertilization in cats. Implementation of the CASA technique in cat reproduction could be interesting to further optimize assisted reproductive techniques in domestic cats and endangered wild felids.

Effect of dilution temperature on boar semen quality.

As boar semen is very sensitive to cold shock and changes in temperature during semen processing can have a profound impact on semen quality, the effect of the extender temperature at the time of dilution was investigated in a two-step dilution protocol for boar semen being processed for liquid storage. Fifteen boars of different breeds and ages from a commercial artificial insemination centre were included. One ejaculate per boar was collected and processed with Beltsville Thawing Solution semen extender. Each ejaculate was diluted (1 : 1) at 30 °C, and subsequently, the samples were diluted (30 × 10(6) sperm/ml) with either preheated extender [29.3 °C ± 0.2 °C, group A (GA)] or extender at room temperature [22.7 °C ± 0.6 °C, group B (GB)]. Samples were transported to the Faculty of Veterinary Medicine (University of Ghent, Belgium) in two isotherm boxes (one per group), stored at 17 °C and investigated for three consecutive days (D0 to D2). At D0, D1 and D2, motility parameters [computer-assisted semen analysis (CASA)] and the per cent of sperm with intact membrane (% IM) by eosin nigrosin staining were evaluated. At D0 and D2, the % of sperm with intact acrosome (% IA) was studied by Pisum sativum agglutinin staining. The average temperature of the 1 : 1 dilution was 29.4 °C ± 1.1 °C immediately after extender addition. No significant differences were found between groups for per cent motility [79.3 ± 9.0 for GA and 81.1 ± 9.2 for GB (p = 0.372)], % progressive motility [56.5 ± 13.3 for GA and 58.4 ± 13.8 for GB (p = 0.737)] or any CASA parameter. No differences were found for % IM [85.1 ± 10.7 and 84.5 ± 3.8 for GA and GB, respectively (p = 0.761)] and % IA [72.2 ± 9.4 for GA and 68.3 ± 16.6 for GB (p = 0.792)]. In conclusion, when a two-step dilution is performed, preheating the extender for the second dilution to match the semen temperature did not result in better semen quality compared to a dilution at a moderate room temperature.

Multiple-locus variable-number tandem-repeat analysis is a suitable tool for differentiation of Mycoplasma hyopneumoniae strains without cultivation.

An assay based on multiple-locus variable-number tandem-repeat analysis allowed differentiating and studying diversity and persistence of Mycoplasma hyopneumoniae strains in pig herds without prior cultivation. The test had a discriminatory index of >0.99 and was applied reliably to porcine bronchoalveolar lavage fluid and tracheal swabs.

The kinetics of the arginine deiminase pathway in the meat starter culture Lactobacillus sakei CTC 494 are pH-dependent.

Lactobacillus sakei is frequently present as the dominant lactic acid bacterium in spontaneously fermented meat products, demonstrating its competitiveness in and adaptation to the meat environment. Since meat is generally low in carbohydrate content, the ability to utilize other energy sources to generate ATP, such as arginine via the arginine deiminase (ADI) pathway, represents a competitive benefit. In this study, the kinetics of growth and arginine conversion capabilities of Lb. sakei CTC 494 were analyzed, and a model was set up to describe the influence of pH on growth and arginine conversion. A series of in vitro batch fermentations using reconstituted MRS medium at different constant pH values (pH 4.50-pH 7.75) was performed. Arginine conversion through the ADI pathway, which was activated from the stationary growth phase on, resulted in the production of both citrulline and ornithine for all pH conditions tested. However, the pattern and the ratio of the end-products of the ADI pathway were influenced by pH. For certain pH values (between pH 5.0 and 6.5), a further conversion of citrulline into ornithine was found when all arginine was depleted. Characterization of responses of the ADI pathway in Lb. sakei CTC 494 to environmental conditions will allow a better understanding and control of this important starter culture in meat fermentations.

Incidence and prevention of early parturition in sows.

A retrospective study, based on 60,990 farrowing records from 35 commercial herds, was performed to determine the incidence of early parturition (<114 d) and to investigate the relationship between early parturition and total number of piglets born, number of piglets born alive and percentage of stillborn piglets per litter. The mean gestation length was 115.4 ± 1.62 d, and early parturition occurred in 10% of all farrowing records. Sows with early parturition had significantly more stillborn piglets and a larger litter size compared to sows with a normal gestation length (114-117 d). Sows with a gestation length <114 d were 1.2 times (95% CI: 1.19-1.21; p<0.001) more likely to have an early parturition at the subsequent parity. A second study was performed in four herds (n=329) to investigate the efficacy of altrenogest administration on 110-112 d (T112) or 111-113 d (T113) of gestation for preventing early parturition and to investigate the effect of this treatment on the reproductive performance of sows. The interval between the last altrenogest treatment and the onset of parturition was 3.3 ± 1.32 (T112) or 2.0 ± 0.89 (T113) days. The gestation length of sows of the altrenogest group (T112 + T113) (115.3 ± 1.23 d) was significantly longer compared to gestation length of the non-treated sows (114.7 ± 1.69d) (p<0.01). Altrenogest treatment had no negative effect on the reproductive performances of the sows. In conclusion, the administration of altrenogest in late gestation is an effective and safe method to prevent early parturition and can counteract the reproductive losses because of premature farrowing, which may occur in a substantial part of the farrowing events.

Dose-response effect of fish oil substitution in parturition feed on erythrocyte membrane characteristics and sow performance.

The present study aimed to investigate whether n-3 polyunsaturated fatty acids (PUFA) incorporate into erythrocyte membranes of peripartal sows in a dose-responsive manner and whether the altered fatty acid profile affects the cell membrane characteristics. At day 109 of gestation (day 0), 51 sows were divided into five treatment groups. Each group received a diet with a different ratio of fish oil to pork lard for nine consecutive days. Blood samples were taken at day 0 and 10 days later. The fatty acid profile of erythrocytes was determined, as well as the osmotic fragility and oxidative stability of erythrocytes. Thiobarbituric acid reactive substances (TBARS) and ferric reducing ability of plasma (FRAP) were determined in plasma samples. Finally, reproductive and performance parameters of both sows and piglets were recorded until weaning. Supplementation of fish oil during the peripartal period changed the fatty acid profile of erythrocyte membranes in a dose-responsive manner. Although the n-3 PUFA content of erythrocyte membranes increased with increasing amounts of fish oil in the diet, no significant effect on erythrocyte osmotic fragility could be recorded. In contrast, oxidative stability of erythrocytes decreased linearly with increasing amounts of fish oil in the diet. Similarly, both TBARS and FRAP linearly increased with increasing percentages of fish oil in the diet. Neither piglet nor sow performance was influenced by dietary treatments, except for a decrease of both piglet survival and weaning weight with increasing quantities of fish oil supplemented. It is concluded that changes in dietary lipid sources can affect the membrane's fatty acid profile within days, and mainly influences oxidative stability of the cells.

Effects of dietary fibre on metabolism and performance in sows.

The etiology of Postpartum dysgalactia syndrome (PDS) includes stress οn preparturition and constipation associated with low water intake or low fiber intake. The aim of this study was to investigate the effects of a raw crude fibre concentrate (Arbocel®) on sow's metabolism and performance. 100 sows from a farm suffering from PDS, were divided into two groups, with equal distribu- tion of their parity (1 to 5 parity): a) T1 group (control group): 50 sows were fed with regular gestation feed (GF), pre-farrowing feed (PFF), and lactation feed (LF), b) T2 group: 50 sows were fed with regular GF, PFF and LF supplemented with topdress Arbocel® from 104th day of gestation until 7th day of lactation). Health parameters [faeces score (FS), PDS score (PDSS), body condition score (BCS)], performance parameters and liter characteristics were recorded. Blood samples were collected from 25 sows / group (5 sows per parity) 24 h after birth of last piglet and on 14th day of lactation for the evaluation of insulin, leptin and ghrelin levels in the serum, using commercial ELISA kits. In T2 group, BCS at farrowing (p⟨0.001), FS (p=0.001) and PDSS (p=0.003) were improved significantly. The number of piglets stillborn and dead due to crushing decreased (p=0.001), while the number of liveborn (p=0.016) and weaned piglets (p=0.001) increased in T2 group. Moreover, in T2 group, the BW of piglets at weaning was higher (p⟨0.001). A significant increase of insulin (p=0.032) and leptin (p=0.032) levels in serum was noticed in T2 group 24 h after farrowing. In conclusion, the supplementation of extra crude fibre in breeding stock with PDS problems due to nutritional imbalance has beneficial effects on their health and performance.

Evaluation of the agreement between Brix refractometry and serum immunoglobulin concentration in neonatal piglets.

Neonatal piglets lack immunoglobulins at birth. Sufficient colostrum intake (CI) and immunoglobulin absorption are essential for an appropriate passive transfer of immunity via the colostrum. Most methods to measure immunoglobulins in serum of piglets are labour-intensive, expensive or imprecise and not designed for on-farm use. The present diagnostic test study evaluated digital Brix refractometry to measure immunoglobulins in serum of neonatal piglets and to suggest thresholds for different serum immunoglobulin concentration. Additionally, agreements between Brix refractometry and optical refractometer (serum total protein, STP) and between Brix refractometry and ELISA (immunoglobulin G, IgG) were also investigated. Forty-five sows and 269 piglets from three different farms were enrolled in the study. Piglets were weighed at birth and 24 h later to calculate the CI. Serum was collected at 24 h after birth and analysed for STP, γ-globulins (electrophoresis), % Brix and IgG. In piglets, median (interquartile range, IQR) CI was 412 (196) g per piglet. Median (IQR) STP, γ-globulin and % Brix concentrations in piglet serum were 60 (11) g/L, 35 (10) g/L and 8 (2) %, respectively. Average (±SD) IgG concentration was 49 ±â€¯23 g/L. Passing-Bablok regression revealed a strong concordance between % Brix and STP (Kendall's tau (Τ): 0.620, P < 0.0001, n = 267) and % Brix and γ-globulin concentration (Kendall's Τ: 0.575, P < 0.0001, n = 267). The agreement between the Brix refractometer and IgG concentration was poor (Kendall's Τ: 0.267, P < 0.0001, n = 269). Receiver operating characteristic curves were performed to evaluate test characteristics of Brix refractometry for three γ-globulin cut-off values, i.e. 10, 20 and 30 g/L. The % Brix cut-off values resulting in the optimal combination of sensitivity and specificity were 5.4 (100 and 98.5%), 7.0 (100 and 89.3%) and 7.9 (90.1 and 80.6%), respectively. In conclusion, digital Brix refractometry is a sufficiently fast and practical method to assess serum γ-globulin concentrations in neonatal piglets on-farm and to evaluate them by considering the thresholds found in this study. Further studies are needed to validate those thresholds regarding piglet's survival in the pre-weaning period.

Short-term exposure to hydrogen peroxide during oocyte maturation improves bovine embryo development.

Recent studies have shown that short-term exposure of oocytes to a stressor such as hydrostatic pressure or osmotic stress might induce stress tolerance in embryos. The aim of the present study was to investigate the consequences of short-term hydrogen peroxide (H(2)O(2)) exposure to bovine in vitro matured cumulus-oocyte complexes (COCs) on subsequent preimplantation embryo development and apoptosis. In the first experiment, mature COCs were incubated in H(2)O(2) at concentrations ranging between 0.01 and 100 micromol/l, and subsequently fertilized and cultured. Oocyte incubation with 50-100 micromol/l of H(2)O(2) resulted in a significantly higher blastocyst yield (47.3%) in comparison with control medium (31.8%), while apoptotic cell ratio was inversely related with H(2)O(2) concentration. In the second experiment, we showed that the stress tolerance after H(2)O(2) exposure was not mediated by increased glutathione content in treated oocytes nor by enhanced fertilization or penetration. Further research should concentrate on the potential role of players that have been associated with stress tolerance in somatic cell lines.

Effect of organic selenium in the diet on sperm quality of boars.

The effect of a diet supplemented with organic selenium (Se) on sperm production and quality of boars was investigated. Sixty mature boars from a commercial artificial insemination centre were randomly allocated at Day (D) 0 into Group A and B. Group A received the regular ration supplemented with inorganic Se (0.4 mg/kg feed as Na(2) SeO(3)) whereas Group B was switched to the same diet but with organic Se (0.4 mg/kg fed as Se-yeast). The sperm was investigated during 4 months (D0, D30, D60, D75, D90, D105 and D120). Sperm concentration and motility were objectively measured using a photometer and Computer Assisted Semen Analysis (CASA) respectively. Morphology of the sperm was assessed using eosin-nigrosin staining and the resistance to induction of oxidative stress (production of malonaldehyde, MDA) through thiobarbituric acid reagent species analysis. Additionally, the Se concentration in sperm and blood plasma were measured. Repeated measures analysis of variance (anova) from D60 to 120 (spermatogenesis of approximately 2 months) or anova at D120 (Se concentrations) were used for statistical analysis. The total number of ejaculated sperm was not significantly different between both groups, but boars of Group B had a significantly higher sperm concentration (434.6 vs 514.1 × 10(6) sperm/ml; p < 0.05). Small differences (p < 0.05) were observed between both groups for some CASA parameters, namely straight line velocity (µm/s) (Group A: 48.3, Group B: 45.1), straightness (%) (Group A: 65.6, Group B: 62.2) and linearity (%) (Group A: 32.2, Group B: 29.3). The sperm of Group B showed more oxidative stress (4.1 vs 4.9 µmol MDA/l; p < 0.05) compared with those of Group A. No significant differences (p > 0.05) were observed for the other parameters. Under the present study conditions, changing from inorganic Se to organic Se in the diet of boars increased sperm concentration but reduced some motility parameters and resistance to oxidative stress.

Combining quantitative and qualitative approaches to determine viability of a potential Salmonella Typhimurium vaccination program in pigs in Belgium.

Vaccinating pigs against Salmonella Typhimurium (ST) might be a way to control ST infections at farm level and reduce human infections. Two main issues have to be addressed before such a mandatory vaccination program can be implemented: the effective reduction of attributable human incidence has to be demonstrated and all socio-economic barriers impacting the attitude and motivation of the pig sector have to be lifted. The present research used a quantitative microbial risk assessment model to estimate the effect of different hypothetical Salmonella spp. and ST mitigation strategies on the annual prevalence of human salmonellosis along the minced pork production chain. In addition, a qualitative study aimed to list the potential concerns of the pig sector about the implementation of a hypothetical future vaccination program. The following themes were the most often mentioned: awareness, vaccine cost-benefit/effectiveness, legislation, monovalent vaccine, time and labour required to vaccinate, vaccine registration and trade restriction. Cost-effectiveness and cost-benefit of vaccination were cited by all the key interviewees (n = 12). However, based on the quantitative microbial risk assessment model, vaccination alone may not be sufficiently effective to reduce the annual human salmonellosis prevalence. A combination of different control measures along the food chain, with a special focus on interventions at the slaughterhouse, might be more effective in achieving the desired goal than vaccination alone.

High biosecurity and welfare standards in fattening pig farms are associated with reduced antimicrobial use.

In order to reduce antimicrobial use in pig production, the consequences of insufficient biosecurity and welfare problems need to be known. This study aimed to investigate associations between the number of antimicrobial treatments per fattening pig, and biosecurity, indicators for animal welfare as well as the prevalence of lesions at slaughter. The data used in this study were extracted from the pig health and welfare classification system (Sikava), which gathers data on medicine usage, meat inspection, animal welfare and the condition of farm buildings from over 95% of pig production in Finland. The data were registered during years from 2011 to 2013. Upon antimicrobial prescription, information on the number of fattening pigs treated and the main reason for treatment was recorded. In addition, at least 4 times per year, pig farms registered in Sikava were visited by the farm veterinarian who assessed, among other things, biosecurity and indicators for animal welfare (air quality, condition of facilities, cleanliness, enrichment and stocking density). Finally, data from slaughterhouse inspections were collected (number of carcasses with joint infection, abscesses, lung lesions, pleurisy and liver lesions). For analysis, these datasets were aggregated at the farm level to a quarter of a year. During the studied period, the mean number of antimicrobial treatments per fattening pig per 3 months was equal to 0.09. The main reasons for antimicrobial treatments were musculoskeletal diseases, tail biting and respiratory disorders (42, 33 and 12% of diagnoses, respectively). The meat inspection scoring indicated that as much as 14.7% of all pigs had pleurisy, 5.3% liver lesions and 4.1% abscesses. A standard zero-inflated negative binomial model was used to identify factors associated with the number of antimicrobial treatments per pig. The count of antimicrobial treatments per pig increased with the size of a farm. Regardless of prevalence of lesions, farms with poor drinking equipment, insufficient enrichment and a combination of poor condition of pens and high stocking density were associated with an increased number of antimicrobial treatments for musculoskeletal diseases per pig. Problems with stocking density and enrichment were associated with the number of antimicrobial treatments for tail biting, although these results depended on prevalence of joint infections. Problems with air quality and the combination of poor cleanliness and poor condition of facilities were associated with increased number of antimicrobial treatments due to respiratory diseases. This study suggests that by improving biosecurity and welfare at pig farms, antimicrobial use can be reduced.

Carcass gain per kg feed intake: developing a stakeholder-driven benchmark for comparing grow-finishing pig performance.

Feed conversion ratio (FCR) in grow-finishing pigs is one of the most important determinants of pig farm profitability and production efficiency. In its simplest form, FCR represents the amount of feed used per unit weight gain of the pig. Yet, this approach entails various limitations hampering its practical applicability such as availability of accurate data and large variation in ways to adapt FCR values for different starting and end weight as well as mortality rates. Various stakeholders are using their own formulas to determine FCR creating a 'definition nonconformity' when comparing FCRs among farms. This study aimed to optimize the calculation of FCR through the use of participatory qualitative research. A multidisciplinary research group of 9 persons (animal scientists, veterinarians and agricultural economists) and a consulting group of 31 stakeholders (representing the Flemish primary sector, feed industry, pharma, genetic companies, large retailers, academia and policy institutions) were involved. The decision problem analysis started with a literature review, followed by 25 in-depth interviews and their analyses (NVivo 11™). This led to an additional literature review and the formation of focus (expert) groups that helped to formulate preliminary FCR formulas. Revision rounds between the research team and the stakeholders further fine-tuned the formulas with the final result being two distinct complimentary formulas that are fit for purpose. Both refer to carcass gain per kg feed intake (plain (CGF) and standardized (CGFstandardized)). The first formula (CGF), namely ${{{\it{number \, delivered \, pigs}} \times {\it{average \, warm \, carcass \, weight}} - {\it{number \, stocked \, piglets}} \times {\it{average \, piglet \, weight}} \times {\it{piglet \, carcass \, yield}}} \over {{\it{feed \, consumption}}}}$ is an objective representation of the animals' performance. The second formula (CGFstandardized) was developed for farm benchmarking, incorporating a seven-step standardization process that corrects for mortality and 'standardizes' for a fixed (yet fictive) live weight trajectory of 25 to 115 kg. This second formula allows to compare farms (or batches of fattening pigs) with different weight trajectories and different mortality rates. A webtool was designed to ease this standardization process (https://varkensloket.be/tools/CGF).

Short Communication: effect of positive handling of sows on litter performance and pre-weaning piglet mortality.

Fear and environmental stressors may negatively affect the welfare of farm animals such as pigs. The present study investigated the effects of music and positive handling on reproductive performance of sows (n = 1014; parity 1 to 8) from a commercial pig farm practicing a batch farrowing system. Every 2 weeks, 56 sows were moved from the gestation unit to conventional-crated farrowing houses 1 week prior to expected farrowing. Treated (T; n = 299) and control (C; n = 715) sows were included in the study. In the farrowing houses, auditory enrichment (music from a radio) was provided to sows of T groups daily from 0600 to 1800 h until the end of lactation. Until the day of farrowing, T sows were additionally subjected, for 15 s per day per sow, to continuous back scratching by one member of farm staff. Litter performance and piglet mortality were recorded and analysed between T and C sows using linear mixed regression models. The number of liveborn piglets (C 13.85 v. T 13.26) and liveborn corrected for fostering (C 13.85 v. T 13.43) was significantly higher (P < 0.05) in C groups compared to the T groups. The number of stillborn piglets was 0.60 and 0.72 in T and C groups, respectively (P > 0.05). With regard to piglet mortality, a linear mixed regression model showed a significant overall effect of treatment in reducing piglet mortality (P < 0.01). Yet, the effect of treatment varied according to litter size (number of liveborn piglets) with a diminishing treatment effect in sows with a high litter size (P < 0.01). Pre-weaning survival was improved in the current study by the combined effect of daily back scratching of sows prior to farrowing and providing music to sows and piglets during lactation. Further research is needed to assess the separate effects of both interventions.