Evaluation of the specificity of three enzyme-linked immunosorbent assays for detection of antibodies against Salmonella in bovine bulk milk

Background

The Swedish Salmonella control program has been running for decades and has resulted in a low prevalence of Salmonella in Swedish food producing animals. Routine bacteriology is used to detect Salmonella, however, bacteriology is time consuming, costly and has a low sensitivity. Different enzyme-linked immunosorbent assays (ELISAs) have been developed for detection of antibodies against Salmonella Dublin and S. Typhimurium in bovine bulk milk, individual milk samples as well as in sera. Screening bulk milk for antibodies against Salmonella spp. could improve the cost-effectiveness of the surveillance in Swedish dairy cattle, but as characteristics of tests may vary in different populations, tests should always be evaluated in the specific population where they will be used. Hence, the aim of this study was to evaluate the specificities of three bovine ELISAs when used to analyse bulk milk samples from Swedish dairy cattle. A second aim was to compare the performance of the two Dublin ELISAs tested.

Methods

Bulk milk samples for analysis were randomly selected from samples collected within the Swedish bulk milk sampling scheme and analyzed with the three ELISAs; a Danish in-house Dublin ELISA, PrioCHECK® Salmonella Ab bovine Dublin ELISA and PrioCHECK® Salmonella Ab bovine ELISA (hereafter named mixed ELISA). The specificities of the ELISAs were calculated assuming a disease-free status in Sweden i.e. that all test positive samples were assumed to be false positive results. This assumption can be used when a disease is known to be infrequent.

Results

The calculated specificities of the two Dublin ELISAs and the mixed ELISA, when using the producer’s recommended cut-off value of the corrected optic-density percent (ODC%) were 99.4% (95% Confidence Interval (CI): 98.8% -99.8%), 99.4% (95% CI: 98.8% -99.8%) and 97.9% (95% CI: 96.8% -98.7%), respectively. The correlation between the ODC% values of the two Dublin ELISAs was 0.83.

Conclusions

We conclude that the evaluated ELISAs have sufficiently high specificities to be used as supplement to bacteriological examinations in the Swedish Salmonella control program in cattle as well as a primary screening test in routine surveillance for S. Dublin.     

Effect of dry period length on the effect of an intramammary teat sealant on the risk of mastitis in cattle treated with antibiotics at drying off

AIM: The aim of this study was to evaluate, under farm conditions, the use of a teat sealant in addition to whole herd dry cow antibiotic therapy on the risk of clinical mastitis in dairy cattle at pasture, and to evaluate the impact of dry period length on that risk and the impact of the teat sealant on that risk.

METHODS: Dairy cows in three herds which used routine whole herd antibiotic therapy were randomly assigned to receive either treatment with an internal teat sealant (n=322) or no additional treatment (n=313) at drying-off between March and May 2010. All clinical mastitis cases during the dry period and to the end of the subsequent lactation were recorded by farm staff; factors affecting risk of clinical mastitis were then analysed using a Cox proportional hazards model.

RESULTS: Median duration of the dry period was 112 days with >25% of cows having a dry period >130 days. The incidence risk of mastitis during lactation for cows treated with teat sealant was 9.9 (95% CI=6.9–13.7) cases per 100 cows compared with 17.9 (95% CI=13.8–22.6) cases per 100 cows for cows treated with antibiotic alone. The addition of a teat sealant to dry cow antibiotic therapy decreased the risk of clinical mastitis only in the first 33 days after calving (Hazard risk 0.24 (95% CI=0.12–0.48)). Length of dry period did not significantly affect the risk of clinical mastitis, or the effect of adding teat sealant to dry cow antibiotic therapy on the risk of clinical mastitis.

CONCLUSIONS: In these herds where, based on the mastitis history, whole herd antibiotic therapy had been recommended, the use of a teat sealant significantly reduced the risk of clinical mastitis. This effect was limited to the first 33 days after calving; subsequently there was no significant effect of treatment. There was no effect of dry period length on risk of clinical mastitis, nor any significant interaction with treatment.

CLINICAL RELEVANCE: Combination therapy with teat sealant and antibiotic was effective under New Zealand conditions in herds using whole herd antibiotic treatment at drying off. Teat sealant reduced risk of clinical mastitis in cattle with dry periods substantially longer than 100 days, and there was no evidence that this effect changed as dry period length increased.     

Pigs weaned from the sow at 10 days of age respond to dietary energy source of manufactured liquid diets and exogenous porcine somatotropin

Previous research indicates that the neonatal pig does not alter feed intake in response to changes in the energy density of manufactured liquid diets. Also, the limited response of IGF-I to exogenous porcine ST (pST) previously observed in young pigs may be influenced by the source of dietary energy. Our objectives were to 1) determine the effect of a high-fat (HF; 25% fat and 4,639 kcal/kg ME; DM basis) or low-fat (LF; 2% fat and 3,481 kcal/kg ME; DM basis) manufactured liquid diet on pig performance; and 2) determine whether the limited response to exogenous pST in young pigs depends on the source of dietary energy. Two replicates of 60 pigs (n = 120; barrows and gilts distributed evenly), with an initial BW of 4,207 +/- 51 g, were weaned from the sow at 10 d of age and used in a randomized complete block design. Pigs were assigned by BW to one of six pens. Diets were formulated to provide a constant lysine:ME ratio and were fed on a pen basis for a duration of 9 d. On d 5, barrows and gilts within a pen were assigned randomly to receive either 0 or 120 microg of pST.kg BW(-1).d(-1) for 4 d. Pigs gained 336 +/- 9 g/d, which resulted in an ending BW of 7,228 +/- 120 g, regardless of dietary treatment (P > 0.15). Pigs fed the LF diet consumed 17% more DM per pen daily than pigs fed the HF diet (2,777 +/- 67 vs. 2,376 +/- 67 g/d, P < 0.01), but calculated ME intake did not differ between dietary treatments (P > 0.20). The G:F was 24% greater in HF- than in LF-fed pigs (P < 0.01). Plasma urea N concentrations were higher in the HF-fed pigs (11.0 +/- 0.6 mg/dL) than in pigs fed the LF diet (6.2 +/- 0.6 mg/dL; P < 0.05). Treatment with pST increased circulating IGF-I (P < 0.01) and decreased PUN (P < 0.01) concentration 32 and 25%, respectively, regardless of dietary treatment (P > 0.30). Circulating leptin averaged 1.8 +/- 0.1 ng/mL and was not affected by dietary treatment (P > 0.35) or pST (P > 0.40). These results suggest that the ST/IGF axis is responsive in the young pig and the increase in circulating IGF-I and growth is independent of the source of dietary energy. Also, young pigs respond to a lower energy density liquid diet with increased feed intake, without altering growth performance, apparently utilizing a mechanism other than circulating leptin.     

When less is more: heterogeneity in grass patch height supports herbivores in counter-intuitive ways

Herbivores are an integral part of the African landscape and have evolved with the vegetation to create the savanna landscape. Managers of these landscapes can benefit from a better understanding of how indigenous herbivores use the landscape to which they are adapted. In this study we observed which patches were frequently utilised, by doing regular monthly road counts, grass height observations and dung counts on selected short grass patches in the Kruger National Park. Smaller-framed impala and blue wildebeest (meso-herbivores) were most regularly seen on these nutritious patches, while from dung deposits it was clear that the even larger-framed buffalo (mega-herbivores) spent time there. This preference can be explained by considering the nutritional needs and food intake of the herbivores. Smaller-framed herbivores seem to be able to satisfy their dietary requirements on the high-quality forage patches, while larger-framed herbivores seem to supplement the quality forage by also spending foraging time on areas of higher grass biomass. From this insight we propose that range management should take herbivore preferences into account and allow herbivores to select and concentrate their foraging on the most nutritious forage. This approach is likely to decrease inputs while allowing animals to maintain or increase production.     

Bone Morphogenetic Protein‐6 (BMP‐6) Stimulates the Antrum Formation by the Regulation of its Signalling Pathway in Caprine Pre‐antral Follicles Cultured In Vitro

BMP‐6 has been found to be important to ovarian cells and oocyte, as well as to uterus. Thus, this study investigated the effect of bone morphogenetic protein (BMP‐6) and recombinant follicle‐stimulating hormone (rFSH) alone or in combination on the in vitro culture (IVC) of isolated caprine secondary follicles (Experiment 1) and the mRNA levels for BMP receptors/Smad signalling pathway (BMPR1A, BMPR2, SMAD1, SMAD4, SMAD5, SMAD6, SMAD7 and SMAD8) in vivo and in vitro using BMP‐6 (Experiment 2). Secondary follicles were cultured in αMEM⁺ alone (control medium) or supplemented with BMP‐6 at 1 or 10 ng/ml and rFSH alone or the combination of both BMP‐6 concentrations and rFSH. The results from Experiment 1 showed that the antrum formation rate was higher in the BMP‐6 at 1 ng/ml (p < 0.05) than in MEM. In Experiment 2, the mRNA expression for BMPR2, SMAD1, SMAD5 and SMAD6 was detected in non‐cultured control and after in vitro culture (MEM and 1 ng/ml BMP‐6); while the expression of SMAD7 and SMAD8 mRNA was only detected after IVC, SMAD4 was only detected in the BMP‐6 at 1 ng/ml treatment. In conclusion, the low BMP‐6 concentration positively influenced antrum formation and ensured normal mRNA expression for BMP receptor and Smads after IVC of caprine secondary follicles.     

Characterization and Liming Effect of Wood Ash Generated from a Biomass-Fueled Commercial Power Plant

Laboratory and greenhouse experiments were carried out to characterize and examine wood ash from a biomass-fueled commercial power plant as a lime. Experimental treatments include wood ash and two commercial limes applied at six application rates. The effective calcium carbonate equivalence (ECCE) of wood ash ranged from 22 to 49%. Wood ash increased soil pH by 0.31–3.18 over unlimed soils, and these changes in pH were similar to the effect of commercial limes. The soil pH at three weeks after wood ash application was constant through week seventeen. Wood ash increased electrical conductivity by 0.13–0.63 dS m^?1 compared to unlimed soils at three weeks after liming; however, a 39–69% decreased was observed by week seventeen. Common bermudagrass did not respond to the application of wood ash and commercial limes. The results of this study suggest that wood ash from the biomass-fueled commercial power plant with low ECCE could be a liming material.     

Determination of Critical Soil Silicon Levels for Rice Production in Louisiana Using Different Extraction Procedures

While silicon (Si) fertilization is widely practiced in rice production, establishing critical soil Si levels has remained understudied. Field trials were established at 12 sites across Louisiana from 2013 to 2015 to determine critical soil Si for rice cultivation. Five silica slag (14% Si) rates at 0, 1, 2, 4, 6, and 8 Mg ha^?1 and two lime rates (2 and 4 Mg ha^?1) were arranged in randomized complete block design with four replications. Post harvest soil samples were analyzed for Si using seven extraction procedures. The critical soil Si levels established by the linear plateau model using 0.5 M acetic acid-1 hr (OAc-1) extraction procedure were 36, 41 and 50 mg kg^?1 for plant Si uptake, grain yield, and relative yield as response variables, respectively. Generally, soils having high initial Si and pH had minimal responses to Si fertilization, whereas Si content of soils with low initial Si was significantly increased.     

Epigallocatechin‐3‐gallate (EGCG) and green tea polyphenols do not improve stallion semen parameters during cooling at 4°C

Stallion semen storage for artificial insemination is mainly based on liquid cooled storage. In many stallions this technique maintains sperm quality for an extended period of time (24–72 hr) at 7°C. While this technique is commonly used in the horse industry, there can be a decline in fertility in some stallions, due to an inability of their sperm to tolerate the cool storage process. The aim of the present work was to evaluate the effect of two natural antioxidants (epigallocatechin‐3‐gallate (EGCG) at 20, 60 and 120 μm and green tea polyphenols, and p at .001, .01 and .1 mg/ml) on some sperm parameters (sperm motility, viability/acrosome integrity and DNA quality) in extended semen immediately after its collection (T0) and after 2, 6, 24 and 48 hr of cool storage. Two ejaculates from three trotter stallions were analysed after 48 hr of storage at 4°C. No beneficial effect on the analysed parameters was observed: the two antioxidants were not able to improve sperm quality after 48 hr of storage. These results are in agreement with previous findings on the effect of different antioxidants reported by other researches, who have demonstrated that stallion semen keeps good antioxidant capacity after dilution for 24 hr. In conclusion, the positive effect exerted by antioxidant molecules in other species is not confirmed in the equine one.