Multiple‐Aetiology Enteric Infections Involving Non‐O157 Shiga Toxin‐Producing Escherichia coli – FoodNet, 2001–2010

We describe multiple‐aetiology infections involving non‐O157 Shiga toxin‐producing Escherichia coli (STEC) identified through laboratory‐based surveillance in nine FoodNet sites from 2001 to 2010. A multiple‐aetiology infection (MEI) was defined as isolation of non‐O157 STEC and laboratory evidence of any of the other nine pathogens under surveillance or isolation of >1 non‐O157 STEC serogroup from the same person within a 7‐day period. We compared exposures of patients with MEI during 2001–2010 with those of patients with single‐aetiology non‐O157 STEC infections (SEI) during 2008–2009 and with those of the FoodNet population from a survey conducted during 2006–2007. In total, 1870 non‐O157 STEC infections were reported; 68 (3.6%) were MEI; 60 included pathogens other than non‐O157 STEC; and eight involved >1 serogroup of non‐O157 STEC. Of the 68 MEI, 21 (31%) were part of six outbreaks. STEC O111 was isolated in 44% of all MEI. Of patients with MEI, 50% had contact with farm animals compared with 29% (P < 0.01) of persons with SEI; this difference was driven by infections involving STEC O111. More patients with non‐outbreak‐associated MEI reported drinking well water (62%) than respondents in a population survey (19%) (P < 0.01). Drinking well water and having contact with animals may be important exposures for MEI, especially those involving STEC O111.     

Effects of forage ensiling and ration fermentation on total mixed ration pH,ruminal fermentation and performance of growing Holstein‐Zebu cross steers

Our objective was to determine the effect of forage ensiling and ration fermentation on total mixed ration pH, ruminal fermentation and animal performance. Thirty Holstein‐Zebu cross steers were allotted to feeding treatments for 188 days in a randomized complete block design including: fresh grass‐total mixed ration (GTMR; pH 4.7), grass silage‐TMR (STMR; pH 4.0) and fermented‐TMR (FTMR; pH 3.5). Average daily gain for STMR was greatest during the first 3 months period; however, that for FTMR tended to be greater than GTMR during the second 3 months period (P < 0.10). During the second period dry matter intake for STMR was the greatest (P < 0.01), but feed conversion ratio (P < 0.01) and cost per gain (P < 0.01) were the least for FTMR. Protein digestibility tended (P < 0.10) to be greater for FTMR than STMR and fat digestibility was greater (P < 0.05) for GTMR and FTMR than STMR. FTMR had less (P < 0.01) ruminal NH₃‐N content than STMR. Total volatile fatty acids post‐feeding was greatest for GTMR (P < 0.01). Ruminal proportions of acetic and butyric acids were greater for FTMR than GTMR (P < 0.05); in contrast, propionic acid was greater for GTMR (P < 0.05). Utilizing silage or total ration fermentation did not negatively impact on ruminal pH. STMR and FTMRs can be used to maintain performance of growing crossbred Holstein steers.     

Lactoferrin concentration and expression in New Zealand cows milked once or twice a day

This study evaluated the concentration and expression of lactoferrin (LF) in cows selected for once a day (OAD) milking compared to twice a day (TAD) milking. Milk samples were collected from the Massey University TAD and OAD herds. Milk traits and expression of LF and insulin‐like growth factor 1 (IGF‐1) were analyzed with a general linear model that included the fixed effects of milking frequency, lactation number, interaction between milking frequency and lactation number, and as covariates proportion of F, heterosis F × J and deviation from the herd median calving date. Cows milked OAD produced milk with higher (p < .01) concentrations of protein and lactose than TAD milked cows. Compared to TAD cows, cows milked OAD had higher expression of the LF gene (1.40 vs. 1.29 folds, p = .03) and the IGF‐1 gene (1.69 vs. 1.48 folds, p = .007). The correlation between the expression of LF gene and the concentration of LF in milk was strong (r = .66 p < .001), but the correlation between the expression of the IGF‐1 gene and LF concentration was stronger (r = .94, p < .001). These results suggest that milking frequency affects the milk composition and expression of milk composition genes at early lactation.     

Effects of partial replacement of barley grain with beet pulp on performance,ruminal fermentation and plasma concentration of metabolites in transition dairy cows

The objective of this study was to determine the effect of partial replacement of barley grain with beet pulp (BP) on dry matter intake (DMI), ruminal fermentation, plasma concentration of metabolites and milk yield of transition dairy cows. Twenty‐four multiparous Holstein cows [735 ± 26 kg of body weights and 3.5 ± 0.05 of body condition score (BCS)] were used in a randomized complete block design. Cows were assigned randomly on day 28 relative to expected parturition date to one of three treatments containing (i) 0% BP, (ii) 25% BP or (iii) 50% BP substituted for barley grain on a DM basis. During the pre‐partum period, DMI and energy intake were greater (P < 0.01) in cows fed the BP diet compared with cows fed the barley grain diet. During the post‐partum period, substituting BP for barley grain caused a response in DMI and energy intake, with the highest amount for the 25% BP diet and lowest for the 50% BP diet (P < 0.01). Milk yield was lowest (P < 0.01) for 50% BP diet than the other treatments. During the post‐partum period, cows fed the 50% BP diets had greater rumen pH, molar proportion of butyrate and acetate: propionate ratio (P < 0.01) in the rumen compared with cows fed the 0% BP diets. In addition, cows fed the BP diets had greater (P < 0.01) plasma β‐hydroxybutyrate and lower plasma glucose (P < 0.05) and blood urinary nitrogen (BUN) (P < 0.01) concentrations than cows fed the barley grain diets. Results showed that substituting BP for barley grain was effective in increasing DMI, but it did not have a significant effect on net energy balance during the post‐partum period. However, replacing BP for barley grain at 50% had adverse effects on DMI, milk yield and metabolic status, as indicated by key blood metabolite concentrations.     

Milk production,plasma metabolite profiles and mammary arterial‐venous differences of milk precursors in early lactation cows milked at different frequencies by an automatic milking system

The objective of this study was to clarify the effect of different milking frequencies under an automatic milking system (AMS) on milk yield, plasma metabolite profiles and mammary arterial‐venous (A‐V) differences of milk precursors by mammary tissues in early lactation cows. Twelve Holstein cows were divided into two and four times milking frequency treatments by AMS after calving to 50 days postpartum. Cows were given a partial mixed ration ad libitum and a concentrate diet at every milking. Dry matter intake increased similarly in both treatments with advancing postpartum days. Milk yield was greater (P < 0.001) by 25% with four times milking, but milk composition was not affected by milking frequency. Body weight change was also not affected by milking frequency. Arterial concentrations of glucose and glutamate were lower (P < 0.05) for four times milking frequency. However, arterial concentration of nonesterified fatty acids did not differ between treatments. Although mammary A‐V differences of plasma concentration for most milk precursors did not differ between treatments, estimated plasma flow was higher (P < 0.05) for four times milking frequency. These results indicate that higher milking frequency may increase mammary uptake of milk precursors, whereas may not affect the extent of fat mobilization of early lactating cows from day 20 postpartum onward.     

Influence of dietary fat source on sow and litter performance,colostrum and milk fatty acid profile in late gestation and lactation

The objective of this study was to investigate the effects of dietary supplementation with various fat sources (3.8–3.9% of diet) during late pregnancy and lactation on the reproductive performance, fatty acids profile in colostrum, milk and serum of sow progeny. A total of 80 multiparous sows were randomly fed a control (adding no oil), palm oil (PO), fish oil (FO) or soybean oil (SO) supplemented diet from 90 days of pregnancy to weaning. Supplementation of FO increased litter size of weak piglets, compared with the control‐fed sows (P < 0.05). Dietary FO and SO supplementation, enhanced the weaning survival rate, litter weaning weight, litter weight gain and fat content in milk (P < 0.05). The highest immunoglobulin (Ig)G and IgM levels in colostrum and milk were observed in the FO group (P < 0.05). Meanwhile, the highest concentration of C22:5 (n‐3) and C22:6 (n‐3) in colostrum, milk and piglet serum was observed in the FO group (P < 0.05). Taken together, dietary inclusion of FO or SO improved growth performance of nursing piglets by increasing milk fat output, and FO consumption by sows might benefit the piglets via increasing n‐3 polyunsaturated fatty acid availability and immunoglobulins (IgG and IgM) secretion.     

Influence of riders’ skill on plasma cortisol levels of horses walking on forest and field trekking courses

The aim of this study was to evaluate the influence of rider's skill on the plasma cortisol levels of trekking horses on two courses, walking on field and forest courses (about 4.5 to 5.1 km each). Three riders of different skills did horse trekking (HT) in a tandem line under a fixed order: advanced‐leading, beginner‐second and intermediate‐last. A total of six horses were used and they experienced all positions in both courses; a total of 12 experiments were done. Blood samples were obtained before HT, immediately after and 2 h after HT. As a control, additional blood samples were obtained from the same horses on non‐riding days. Irrespective of the course and the rider's skill, the cortisol level before HT was higher than that of control (P < 0.05). In both courses, the cortisol levels immediately after HT ridden by the advanced rider were higher than that of control (P < 0.05). However, in every case, the cortisol level 2 h after HT was closely similar to the level of the control. Thus, we concluded the stress of trekking horse was not sufficient to disturb the circadian rhythm of the cortisol level, irrespective of the course and the rider's skill.     

Changes in some blood parameters,milk composition and yield of buffaloes (Bubalus bubalis) during the transition period

In this study changes in hematochemical parameters, milk composition and yield were investigated in buffaloes during the transition period. A total of 93 buffaloes 113.9 ± 8.03 months old and 535 ± 50 kg average body weight were used. Parity was recorded, blood samples were collected from 80 days pre‐partum until 70 days post‐partum; milk samples were collected from 5 days to 70 days post‐partum. On serum samples, the values of non‐esterified fatty acids, β‐hydroxybutyrate, glucose, cholesterol, triglycerides, aspartate aminotransferase, alanine aminotransferase, gamma‐glutamyltransferase, urea, total proteins were evaluated. On milk, percentage of fat, protein and lactose, along with the somatic cell count (SCC), milk yield and daily milk production (DMP) were assessed. The peripartum period significantly (P ≤ 0.01) influenced all studied parameters with the exception of glucose. Milk fat percentage showed decreasing trend from 10 until 40 days post‐partum; DMP significantly (P ≤ 0.01) increased from 1 day post‐partum until 40 days post‐partum. Milk yield significantly (P ≤ 0.01) decreased in animals over the sixth lactation. Our results confirmed the importance of transit period in buffaloes. Blood parameters and milk composition alterations are crucial to predict the energy balance status of buffaloes in order to improve their management and feed intake during the transition period.     

Effects of forage offering method on performance,rumen fermentation,nutrient digestibility and nutritional behaviour in Holstein dairy calves

The potential effect of dietary forage supplementation on the performance and rumen development in dairy calves is well established. However, limited research has been directed to the comparative effects of forage offering methods on calf performance. The objective of the present study was to determine the effects of forage provision methods (total mixed ration or free choice) on the performance, nutrient digestibility, rumen fermentation and nutritional behaviour in newborn calves. Forty‐five Holstein dairy calves (3 days of age and 41 ± 2 kg of body weight) were assigned to the following three groups (n = 15): (i) starter without forage provision (CON), (ii) starter supplemented with 10% alfalfa hay (AH) as a total mixed ration (AH‐TMR) and (iii) starter and AH as a free‐choice provision (AH‐FC) for a period of 70 days. All the calves were offered 5 l of milk/day from day 3 to 50, and 2.5 l/day from day 50 until weaning on day 56. Dry matter intake (DMI) was greater (p < 0.01) in the AH‐TMR and AH‐FC treatments than in the CON during the pre‐ and post‐weaning periods. Calves fed the AH‐FC diet showed the highest post‐weaning DMI among the treatments. The calves receiving ad libitum forage tended (p = 0.08) to increase crude protein digestibility and overall volatile fatty acids (VFA) concentrations in the rumen. No differences were observed among the treatments at the time spent on standing, lying, eating and performing non‐nutritive oral behaviours. Compared to CON calves, animals in the AH‐TMR treatment spent more time (p < 0.05) ruminating. In conclusion, our data suggest that forage supplementation in both forage offering methods increased total DMI, ruminal pH and ruminating time in dairy calves. Hence, there is no benefit in the free‐choice provision of AH in dairy calves.     

Milk yield estimation of Ogaden cattle breed based on methods of weigh–suckle–weigh and calves’ growth

Daily milk yield of Ogaden cattle was estimated from milk suckled by 269 calves born and maintained between 1994 and 2004 at Haramaya University Beef Farm, Ethiopia. The weigh–suckle–weigh method and the growth of calves were used to estimate the yield. Mean predicted daily milk yield of the breed based on the weigh–suckle–weigh method was 4.39 ± 0.03 kg day⁻¹. Daily milk yield estimated from the growth of calves up to 3 months of age was 4.59 ± 1.43 kg day⁻¹. The milk yield was significantly affected by seasons of calving (p < 0.001), parities of the dams (p < 0.05), weeks of lactation (p < 0.001) and birth weights of calves (p < 0.05). A relatively higher milk yield was observed from cows calved during the long rain season. A higher milk yield was produced by cows in fifth parity. Heavier calves at birth suckled more milk than lighter calves. Daily milk yield was significantly higher in 3–6 weeks of lactation. The result of this study indicated that the milk yield produced by Ogaden breed was relatively higher than the yield of some Zebu breed measured by partial suckling system in Ethiopia and was comparable to the yield from Zebu cattle in the tropics estimated by the same method.     

A case study of improving milking cow performance and milking system performance with using a flow simulator

Efforts to improve dairy performance have been focused on increasing milk productivity of cows through improved feeding systems and genetic potential. However, methods for evaluating milking system performance based on milk productivity have not yet been established. Milking system performance was evaluated by measuring the claw vacuum at five flow rates (1.9–8.7 kg/min) produced using a flow simulator for a single eight‐swing milking parlor with a high‐line system. Based on these results, a double eight‐parallel milking parlor with a low‐line system was installed and tested. Farmers can take data obtained from evaluations of milking system performance into account for future management decisions, such as renewing the milking system. By renewing the milking system, average milking productivity, somatic cell linear score (LS) of bulk milk, and LS of each cow were significantly improved in the year after installing the new system (p < .01). In addition to checking conventional milking systems, this novel diagnostic method using a flow simulator can be used for checking new installations and also for proposing renovations.     

Variations in protein and fat contents and their fractions in milk from two species fed different forages

This study aimed at determining the variations in milk constituents which could be varied by feed and animal species. To achieve this goal, two groups of homoparity Baladi cows and Egyptian buffaloes (n = 20 per species) were used. Each group was divided into two subgroups (n = 10): subgroup I received legume forage (Egyptian clover) and subgroup II received grass forage (sorghum forage). All experimental animals were fed the diet consisting of concentrate, forage and rice straw as 50, 25 and 25% of dry matter intake respectively. Milk samples were taken for analysis. The trial lasted until the 3rd month of parturition. The main results indicated that lactating cattle fed legume forage significantly (p ≤ 0.01) had more content of casein nitrogen (513 mg/100 ml milk), lower content of glutamic acid (23.56 g/100 g milk protein) and more content of cis‐9, trans‐11 18:2 conjugated linoleic acid (CLA) (0.77 g/100 g milk fat) compared with 433, 26.67 and 0.53, respectively, for cattle fed grass forage. With regard to the species effect, results showed that buffalo milk appeared to contain significantly higher (p ≤ 0.01) contents of casein nitrogen, phenylalanine, glutamic and arachidonic acid compared with cow's milk. However, the latter was significantly (p ≤ 0.01) more in the cis‐9, trans‐11CLA (0.59 g/100 g milk fat) than that in buffalo milk (0.47 g/100 g milk fat). The results revealed that not only forage type played a critical role in determining the variations of milk nitrogen distribution, milk amino acids and fatty acids but also animal species had a significant effect on these parameters.     

Influence of the calf presence during milking on dairy performance,milk fatty acid composition,lipolysis and cheese composition in Salers cows during winter and grazing seasons

The milking of Salers cows requires the presence of the calf. The removal of the calf would simplify the milking routine, but it could also modify the milk yield and the milk and cheese composition. Therefore, the aim of this experiment was to evaluate the effect of calf presence during milking during sampling period (winter or grazing periods), on dairy performance, milk fatty acid (FA) composition, lipolysis and cheese yield and composition. Nine and 8 Salers lactating cows were milked in the presence (CP) or absence (CA) of their calves respectively. During winter, the cows were fed a hay‐based diet and then they only grazed a grassland pasture. Calf presence during milking increased milk yield and milk 16:0 concentration and decreased milk fat content and milk total odd‐ and branched‐chain FA (OBCFA) concentrations. Calf presence only increased initial lipolysis in milk collected during the winter season. Milk from CP cows compared to CA cows resulted in a lower cheese yield and ripened cheeses with lower fat content. Milk from the grazing season had lower saturated medium‐chain FA and OBCFA concentrations and higher 18:0, cis‐9‐18:1, trans‐11‐18:1 and cis‐9, trans‐11‐CLA concentrations than that from the winter season. Initial milk lipolysis was higher in the winter than in the grazing season. These variations could be due to seasonal changes in the basal diet. Furthermore, the effect of calf presence during milking on milk fat composition was lower than that on dairy performance, cheese yield and composition. Removing the calf during the milking of Salers cows seems feasible without a decrease in milked milk, and with a positive effect on cheese yield and fat content, under the condition that we are able to select cows having the capacity to be milked easily without the calf.     

Effect of dietary conjugated linoleic acid on vitamin A status of lactating rats and their offspring

The absorption and metabolism of vitamin A is linked with that of lipids. It is known that conjugated linoleic acid (CLA) affects the lipid metabolism in growing and lactating animals. In the present study, the hypothesis was investigated that dietary CLA influences vitamin A status of lactating rats and their pups during the suckling period. For this purpose, Wistar Han rats were fed either a control diet (control group, n = 14) or a diet containing 0.87% of cis‐9, trans‐11 and trans‐10, cis‐12 (1:1) CLA (CLA group, n = 14) during pregnancy and lactation. Vitamin A concentrations in various body tissues were determined 14 days after delivery in dams and 1, 7 and 14 days after birth in pups, and expression of selected genes involved in metabolism of retinoids was determined in dams. Vitamin A concentrations in liver, plasma and muscle were similar in control and CLA‐fed dams. Expression of genes involved in retinoid transport, storage and degradation in liver and adipose tissue in dams was also not different between control and CLA‐fed dams. Vitamin A concentrations in milk curd, sampled at d 1, 7 and 14 of lactation were not different between control and CLA‐fed dams. Vitamin A concentrations in liver, lung and adipose tissue were also not different in pups from control dams and pups from CLA‐fed dams. In conclusion, we show for the first time that dietary CLA has little effect on vitamin A concentrations and vitamin A metabolism in lactating rat dams and, moreover, does not influence tissue vitamin A concentrations in their newborn and suckling pups.     

The Ability of the Enzyme‐Linked Immunosorbent Assay to Detect Antibody against Staphylococcus aureus in Milk following Experimental Intramammary Infection

Changes in the milk antibody levels against Staphylococcus aureus were measured at the start of an experimental intramammary instillation of either S. aureus (Study I) or Staphylococcus hyicus (Study II). A commercial enzyme‐linked immunosorbent assay system was used. Twenty‐one Holstein cows were enrolled in Study I and 15 Holstein cows were used in Study II. Pathogen instillation began 21 days before the start of the non‐lactating period. Cows received intramammary antibiotic treatment in all quarters immediately after the last milking, the start of the non‐lactating period. Lacteal secretions were collected before the start of the non‐lactating period, and during the immediate postpartum period in both studies, and during the non‐lactating period in Study I. Milk was cultured for mastitis pathogens and S. aureus antibody levels and somatic cell counts were determined from all samples. There was an approximate 2‐week delay in the elevation in antibody levels in response to the instillation of S. aureus. Antibody levels remained elevated in cows with S. aureus intramammary infections postpartum, but were below threshold in cows where intramammary infections were cured during the non‐lactating period. Antibody levels were elevated by S. hyicus intramammary infections, remained elevated for the first 12 days postpartum, but were below threshold by day 21 postpartum. Cows with incipient intramammary S. aureus infections might be misclassified as false negatives by the antibody test. However, results suggest that cows with S. hyicus intramammary infections that were not cured would not be misclassified if milk is withheld from test for the first 30 days postpartum, as recommended by the manufacturer of the test.     

Effects of ear corn silage supplementation on milk production and milk fatty acid profiles in grazing dairy farms

This study investigated the effects of substituting ear corn silage (ECS) for commercial formula feed on milk production and milk fatty acid profiles in grazing dairy farms during the summer season. A field survey was conducted on five grazing dairy farms in every summer month of 2017, 2018, and 2019. Three of the five farms substituted fresh ECS for the commercial formula feed at a ratio of 2:1 from July of each year (ECS farms). Other farms maintained the same feeding management as before (non‐ECS farms). An interview survey was conducted on each farm to calculate feed intake and milk yield per cow. Feed and milk samples were collected in each survey. Milk compositions and milk fatty acid profiles were determined. The substitution of ECS for the commercial formula feed did not affect milk yield or milk composition, but ECS farms maintained low levels of milk urea compared with non‐ECS farms (p < .01). The ECS substitution also influenced some of the milk fatty acid proportions; C16:0 and C16:1 increased, and trans‐11 C18:1, cis‐9,trans‐11 C18:2, and the sum of polyunsaturated fatty acids decreased, while these fatty acid proportions were maintained in non‐ECS farms throughout the summer season (p < .05).     

Impact of α‐amylase supplementation on energy balance and performance of high‐yielding dairy cows on moderate starch feeding

In dairy cows, exogenous α‐amylase is suggested to improve starch utilization and positively affect performance and health traits linked to energy balance and fertility. In a 1‐year feeding experiment, 421 cows were orally supplemented with α‐amylase (treatment: 12.5 g/kg dry matter (DM) addition rate to a concentrated feed) or non‐supplemented (control) on the basis of an ad libitum total mixed ration (TMR). Every cow was allocated to a high‐ (≥32 kg milk/day) or late‐lactation group (<32 kg milk/day), in which the TMR starch content was 220 ± 20.8 g/kg DM and 183 ± 24.8 g/kg DM, respectively. The energetic effect of α‐amylase supplementation seemed to be exclusively related to the high‐lactation stage (5–100 days in milk) in primiparous cows, where the daily milk yield was 32 ± 0.49 versus 31 ± 0.50 kg per cow in the treatment versus control group (P < 0.05). The pluriparous cows did not benefit from the supplementation that way. In neither primiparous nor pluriparous cows, was the milk composition, the fat‐to‐protein ratio, the somatic cell score, the backfat thickness, serum total bilirubin, β‐hydroxybutyrate and the fertility found to be systematically affected by α‐amylase supplementation.     

Effect of vacuum level on milking efficiency, somatic cell counts (SCC) and teat end wall thickness in ewes of Greek mountain Boutsiko breed

The aim of this work was to determine the effect of vacuum level of machine milking on milk yield, milk composition, milking efficiency, somatic cells counts (SCC), and teat end thickness of the Greek mountain breed Boutsiko ewes. Seventy two ewes after a suckling period of 45 ± 5 days were divided into three equal groups of 24 animals for level of milk production, prolificacy and lactation number. The overall experiment lasted 14 weeks. All groups were milked with a pulsation rate of 120/min, pulsator ratio of 50/50, and three vacuum levels of 38, 44 and 50 kPa, respectively, were applied for the three groups. Every two weeks, in a.m. and p.m. milking, measurements were taken for machine milk (MM) and machine stripped milk (MSM), milk composition, SCC, milk emission characteristics (only in a.m. milking) and teat end wall thickness (TEWT) before and after milking. The results have shown that the lower vacuum level of 38 kPa increased (P < 0.05) the percentage of machine milk (82.9%). There were no significant differences among the groups for milk composition, milk flow rate and SCC, but there was a difference in log SCC (P < 0.05) between 38 and 50 kPa. The teat end wall thickness was lower at the lower vacuum level (38 kPa) after milking, and increased (P < 0.001) as the vacuum level was raised to 50 kPa. The findings of this study suggest that with a lower vacuum level of 38 kPa, the ewes are milked better than those with 44 or 50 kPa. Also, the irritation of teats is considerably reduced, while no differences were seen regarding the health of the udder.     

Trans10, cis12 conjugated linoleic acid increases triacylglycerol accumulation in goat mammary epithelial cells in vitro

Trans10, cis12 conjugated linoleic acid (t10c12‐CLA) is well‐established in decreasing milk fat content and causing milk fat depression (MFD) in dairy cattle and goats. However, the detailed mechanisms of its effect are not completely understood. Therefore, we used goat mammary epithelial cells (GMECs) to further study the molecular mechanisms whereby t10c12‐CLA regulates milk fat synthesis. The optimal concentration of t10c12‐CLA (100 μmol/L) for cell culture was determined through a cell vitality and morphology assay, and evaluation of abundance of apoptosis‐related proteins. Oil red O stain indicated that t10c12‐CLA increased concentration of cytoplasmic lipid droplets. Furthermore, t10c12‐CLA increased the intracellular triacylglycerol (TG) content (P < 0.05). Among 16 genes related to lipid metabolism that were measured by quantitative real‐time PCR, t10c12‐CLA down‐regulated (P < 0.05) genes involved in de novo fatty acid synthesis including FASN, ACACA and SCD1, and also down‐regulated the protein expression of FASN and SCD1 but up‐regulated (P < 0.05) the expression of CD36 and ADRP. Overall, the data indicate that a side effect of de novo fatty acid synthesis inhibition by t10c12‐CLA is the up‐regulation of fatty acid uptake and accumulation of lipid droplets in GMECs. The biologic reason for such an effect merits further study.     

Experimental Neospora Caninum Infection in Pregnant Dairy Heifers Raises Concentrations of Pregnancy‐Associated Glycoproteins 1 and 2 in Foetal Fluids

Plasma concentrations of PAG‐1 are used for pregnancy diagnosis and as a marker of placental/foetal well‐being, while those of PAG‐2 may be an indicator of abortion risk in Neospora caninum‐infected cows. Studies have shown that N. caninum infection modifies PAG‐1 and PAG‐2 patterns in maternal blood plasma. However, no prior work has examined the effects of N. caninum infection on concentrations of PAGs in foetal fluids. In this study, PAG‐1, PAG‐2 and pH levels were determined in the amniotic and allantoic fluids of foetuses collected at 152 days of gestation from control uninfected dams and from dams experimentally infected with N. caninum on Day 110 of gestation. Foetal fluids from infected foetuses had significantly higher PAG‐2 concentrations (p = 0.026) and pH values (p = 0.02) than fluids from non‐infected foetuses. In infected foetuses, significantly higher concentrations of PAG‐1 (p < 0.001) and PAG‐2 (p < 0.001) were detected in fluid samples showing antibodies against N. caninum than those without antibodies. Moreover, pH values were significantly higher (p = 0.011) in foetal fluid samples with antibodies than in samples from non‐infected foetuses. In conclusion, this is the first report on the effect of N. caninum infection on PAG levels in foetal fluids. Our results indicate that following the experimental infection of dams with N. caninum on Day 110 of gestation, foetal fluids collected from the infected foetuses of these dams featured higher PAG‐1 and PAG‐2 levels and pH values than fluids from non‐infected controls, provided that the samples tested showed the presence of antibodies. The clinical implications of these findings are that following infection with N. caninum, most cows will experience some level of placental damage and that this injury correlates with foetal fluid PAG levels and pH.