Systematic assessment of the reliability of quantitative PCR assays targeting IS900 for the detection of Mycobacterium avium ssp. paratuberculosis presence in animal and environmental samples.

Mycobacterium avium ssp. paratuberculosis (MAP) is the bacterium responsible for causing Johne's disease (JD), which is endemic to dairy cattle and also implicated in the etiology of Crohn's disease. The difficulty in diagnosing asymptomatic cows for JD makes this disease hard to control. Johne's disease is considered a priority under the One Health approach to prevent the spread of the causative agent to humans. Environmental screening is a strategic approach aimed at identifying dairy herds with animals infected with MAP. It serves as the initial step toward implementing more intensive actions to control the disease. Quantitative PCR (qPCR) technology is widely used for diagnosis. Given that genome sequencing is now much more accessible than ever before, it is possible to target regions of the MAP genome that allow for the greatest diagnostic sensitivity and specificity. The aim of this study was to identify among the published qPCR assays targeting IS900 the more cost-effective options to detect MAP and to validate them in the diagnostic context of JD. Mycobacterium avium ssp. paratuberculosis IS900 is a prime target because it is a multicopy genetic element. A total of 136 publications have reported on the use of IS900 qPCR assays over the past 3 decades. Among these records, 29 used the SYBR Green chemistry, and 107 used TaqMan technology. Aside from the 9 reports using commercial assays, 72 TaqMan reports cited previously published work, leaving us with 27 TaqMan qPCR designs. Upon closer examination, 5 TaqMan designs contained mismatches in primer or probe sequences. Additionally, others exhibited high similarity to environmental microorganisms or non-MAP mycobacteria. We assessed the performance of 6 IS900 qPCR designs and their sensitivity when applied to clinical or environmental samples, which varied from 4 to 56 fold overall. Additionally, we provide recommendations for testing clinical and environmental samples, as certain strategies used previously should be avoided due to poor qPCR design (e.g., the presence of mismatches) or a lack of specificity.

Effects of photoperiod modulation and melatonin feeding around drying-off on bovine mammary gland involution.

The risk for a dairy cow to acquire new intramammary infections is high during the transition from lactation to the dry period, because of udder engorgement and altered immune functions. Once the gland is fully involuted, it becomes much more resistant to intramammary infections. Therefore, strategies to depress milk yield before drying-off and accelerate the involution process after drying-off could be beneficial for udder health. The objective of this study was to assess the effect of photoperiod manipulation and melatonin feeding from 14 d before to 14 d after drying-off on the speed of the involution process. Thirty Holstein cows in late lactation were randomly allocated to one of the following treatments: (1) a long-day photoperiod (16 h of light: 8 h of darkness), (2) a short-day photoperiod (8 h of light: 16 h of darkness), and (3) a long-day photoperiod supplemented by melatonin feeding (4 mg/kg of body weight). Milk and blood samples were collected on d -26, -19, -12, -5, -1, 1, 3, 5, 7, 10, and 14 relative to the last milking to determine concentrations of mammary gland involution markers and serum prolactin. Additional blood samples were taken around milking on d -15, before the start of the treatments, and on d -1, before drying-off, to evaluate the treatment effects on milking-induced prolactin release. The short-day photoperiod slightly decreased milk production and basal prolactin secretion during the dry period. The milking-induced prolactin surge was smaller on d -1 than on d -15 regardless of the treatments. Lactoferrin concentration, somatic cell count, and BSA concentration as well as matrix metalloproteinase-2 and -9 activities increased in mammary secretions during the first 2 wk of the dry period, whereas milk citrate concentration and the citrate:lactoferrin molar ratio decreased. The rates of change of these parameters were not significantly affected by the treatments. The long-day photoperiod supplemented by melatonin feeding did not affect milk production, prolactin secretion, or mammary gland involution. Under the conditions in this study, photoperiod modulation and melatonin feeding did not appear to affect the rate of mammary gland involution.

Effect of inhibiting the lactogenic signal at calving on milk production and metabolic and immune perturbations in dairy cows.

During the periparturient period, the abrupt increase in energy demand for milk production often induces metabolic and immunological disturbances in dairy cows. Our previous work has shown that reducing milk output by milking once a day or incompletely in the first few days of lactation reduces these disturbances. The aim of this study was to reduce metabolic and immunological disturbances by limiting milk production during the first week of lactation by inhibiting the lactogenic signal driven by prolactin. Twenty-two fresh cows received 8 i.m. injections of the prolactin-release inhibitor quinagolide (QUIN; 2 mg) or water as a control (CTL). The first injection was given just after calving, and the subsequent 7 injections were given every 12 h. Milk production was measured until d 28 after calving. Blood samples were taken from d 1 (calving) to d 5 and then on d 7, 10, 14, 21, and 28 to measure concentrations of urea, phosphorus, calcium, glucose, nonesterified fatty acids (NEFA), ß-hydroxybutyrate, and prolactin. Other blood samples were taken on d 2, 5, 10, and 28 to analyze oxidative burst, phagocytosis, and the effect of the serum on the lymphoproliferation of peripheral blood mononuclear cells from donor cows. Blood prolactin concentration was lower from d 2 to 5 but higher from d 10 to 28 in the QUIN cows than in the CTL cows. Milk production was lower from d 2 to 6 in the QUIN cows than in the CTL cows (24.3 ± 6.4 and 34.8 ± 4.1 kg/d on average, respectively). We observed no residual effect of quinagolide on milk production after d 6. During the first week of lactation, blood glucose and calcium concentrations were higher and ß-hydroxybutyrate concentration was lower in the QUIN cows than in the CTL cows. Blood NEFA, urea, and phosphorus concentrations were not affected by the treatment. At d 2 and 5, the phagocytosis ability of polymorphonuclear leukocytes was not affected by treatment; however, quinagolide injection enhanced the proportion of cells that entered oxidative burst, The mitogen-induced proliferation of peripheral blood mononuclear cells was greater when they were incubated with serum harvested from the CTL cows and was negatively correlated with the NEFA concentration in the serum. Reducing the prolactin peak at calving was effective in reducing milk production during the first week of lactation without compromising the dairy cow's overall productivity. Slowing the increase in milk production allowed a more gradual transition from pregnancy to lactation and led to a reduction in metabolic stress and an improvement in some immune system aspects during this period.

Relationship between glucocorticoids and prolactin during mammary gland stimulation in dairy cows.

The objectives of this study were to determine the role of glucocorticoids in the regulation of prolactin (PRL) release induced by mammary gland stimulation and to investigate whether the milk depression induced by glucocorticoids in dairy cows is due to a decrease in PRL release. In experiment 1, 8 dairy cows were used in a 4 × 4 Latin square design. Four hours after the morning milking, the cows received 1 of the following treatments: (1) a 5-min manual stimulation of the mammary gland; (2) an i.v. injection of 1 mg of dexamethasone; (3) 2 infusions of 2.5 g of metyrapone (an inhibitor of cortisol biosynthesis) in the omasum 4 and 2 h before a 5-min stimulation of the mammary gland; or (4) no treatment. Sixty minutes later, the mammary gland of each cow was stimulated for 5 min. Blood samples were collected from 20 min before to 120 min after the start of the treatment. When the mammary gland was stimulated twice in 60 min, less PRL and cortisol were released during the second stimulation. Metyrapone did not affect PRL or cortisol release. Dexamethasone decreased serum cortisol concentration but did not affect PRL concentration. In experiment 2, 16 cows were used in a crossover experimental design consisting of 2 experimental weeks separated by 1 resting week. During the first week, cows were treated as follows: (1) 4 cows were injected with 0.5 g of domperidone (a PRL secretagogue) in canola oil on d 1 and 2 and 20 mg of dexamethasone on d 1; (2) 4 cows were injected with 0.5 g of domperidone on d 1 and 2; (3) 4 cows were injected with canola oil on d 1 and 2 and with 20 mg of dexamethasone on d 1; and (4) 4 cows were injected with canola oil on d 1 and 2. During the second experimental week, the same 4 treatments were repeated, except the cows that did not receive dexamethasone in the first week received it on d 1 of the second week, and cows that did receive it in the first week did not receive it in the second week. On d 1 and 2 of each week, blood samples were collected during morning milking for PRL determination. Dexamethasone reduced milk production and decreased both basal and milking-induced PRL release. It also increased milk fat and protein percentages and decreased milk lactose content. Domperidone increased basal PRL levels in serum and milk but did not affect milk yield. Although we cannot rule out the possibility that inhibition of PRL secretion or reduction of mammary gland PRL responsiveness play a role in the inhibition of milk production by glucocorticoids, the fact that enhancement of PRL secretion by domperidone could not prevent the depression of milk yield suggests that other mechanisms are involved.

Effect of reducing milk production using a prolactin-release inhibitor or a glucocorticoid on metabolism and immune functions in cows subjected to acute nutritional stress.

When cows are unable to consume enough feed to support milk production, they often fall into severe negative energy balance. This leads to a weakened immune system and increases their susceptibility to infectious diseases. Reducing the milk production of cows subjected to acute nutritional stress decreases their energy deficit. The aim of this study was to compare the effects on metabolism and immune function of reducing milk production using quinagolide (a prolactin-release inhibitor) or dexamethasone in feed-restricted cows. A total of 23 cows in early/mid-lactation were fed for 5 d at 55.9% of their previous dry matter intake to subject them to acute nutritional stress. After 1 d of feed restriction and for 4 d afterward (d 2 to 5), cows received twice-daily i.m. injections of water (control group; n=8), 2mg of quinagolide (QN group; n=7), or water after a first injection of 20mg of dexamethasone (DEX group; n=8). Feed restriction decreased milk production, but the decrease was greater in the QN and DEX cows than in the control cows on d 2 and 3. As expected, feed restriction reduced the energy balance, but the reduction was lower in the QN cows than in the control cows. Feed restriction decreased plasma glucose concentration and increased plasma nonesterified fatty acid (NEFA) and ß-hydroxybutyrate (BHB) concentrations. The QN cows had higher glucose concentration and lower BHB concentration than the control cows. The NEFA concentration was also lower in the QN cows than in the control cows on d 2. Dexamethasone injection induced transient hyperglycemia concomitant with a reduction in milk lactose concentration; it also decreased BHB concentration and decreased NEFA initially but increased it later. Feed restriction and quinagolide injections did not affect the blood concentration or activity of polymorphonuclear leukocytes (PMN), whereas dexamethasone injection increased PMN blood concentration but decreased the proportion of PMN capable of inducing oxidative burst. Incubation of peripheral blood mononuclear cells in serum harvested on d 2 of the restriction period reduced their ability to react to mitogen-induced proliferation, and injection of quinagolide or dexamethasone could not alleviate this effect. This experiment shows that prolactin-release inhibition could be an alternative to dexamethasone for reducing milk production and energy deficit in cows under acute nutritional stress, without disturbing immune function.

New insights into the importance of prolactin in dairy ruminants.

In most mammals, prolactin (PRL) is essential for maintaining lactation, and the suppression of PRL inhibits lactation. However, the involvement of PRL in the control of ruminant lactation is less clear, because inconsistent effects on milk yield have been observed with the short-term suppression of PRL by bromocriptine. Therefore, several experiments have been conducted to assess the galactopoietic role of PRL. In an initial experiment, cows in early lactation received daily injections of the dopamine agonist quinagolide for 9 wk. Quinagolide reduced milking-induced PRL release and caused a faster decline in milk production. Quinagolide also reduced mammary epithelial cell activity, survival, and proliferation. In goats, cabergoline, another dopamine agonist, caused a 28% decrease in milk yield the day after injection. In another experiment, cows were injected for 5d with quinagolide, with quinagolide plus bovine PRL injected at milking time, or with vehicles only. Again, quinagolide reduced milk, protein, and lactose yields. Although PRL injections were not sufficient to restore milk yield, they tended to increase milk protein and lactose yields and increased the viability of mammary epithelial cells purified from milk. Recently, our team stimulated PRL secretion with daily injections of the dopamine antagonist domperidone for 5 wk. Milk production increased gradually and was greater in domperidone-treated cows during the last 4 wk of the treatment period. In most experiments where PRL secretion was manipulated, feed intake paralleled the changes of PRL concentration, supporting the idea that PRL increases feed intake to provide the nutrients necessary to support lactation in dairy ruminants. In late-lactation cows, quinagolide and cabergoline decreased milk production within the first day of treatment and induced more rapid changes in several markers of mammary gland involution after drying-off. In addition, quinagolide improved the resistance to intramammary infection, suggesting that PRL inhibition could be an alternative strategy for facilitating drying-off. Prolactin appears to directly affect mammary gland functions, but mammary gland responsiveness to PRL appears to be modulated by local and systemic factors. Therefore, the modulation of the number and isoforms of the PRL receptors as well as the expression of intracellular modulators of cell signaling in the mammary gland require further investigation. In conclusion, these data, combined with those from other studies, provide a good body of evidence that PRL is galactopoietic in dairy ruminants.

The dopamine antagonist domperidone increases prolactin concentration and enhances milk production in dairy cows.

In previous studies, our team showed that the inhibition of prolactin (PRL) secretion by the dopamine agonist quinagolide reduces milk production in dairy cows. The objective of this study was to determine the effects of administration of a dopamine antagonist on basal and milking-induced PRL concentrations in blood and on milk production during positive energy balance and feed restriction in dairy cows. Eighteen mid-lactation Holstein cows received daily s.c. injections of either domperidone (300 mg, DOMP, n=9) or the vehicle, canola oil (CTL, n=9), for 5 wk. During wk 5, all cows were fed at 65% of their dry matter intake in the previous week. Blood and milk samples were collected before (for blood) and during (for milk) the a.m. milking thrice weekly from d -9 to 41 (8d after the last injection). In addition, blood samples were collected during the a.m. milking on d -1 (before the first injection), and on d 1, 28, and 34. Basal PRL concentration was similar in both groups before the start of the treatments. Domperidone injections caused a gradual increase in basal PRL concentration. Feed restriction reduced basal PRL concentration in both the CTL and DOMP cows, but PRL concentration remained higher in the DOMP cows. Prolactin concentration remained elevated in the DOMP cows 7d after the last injection. The milk concentration of PRL increased during the DOMP treatment, but the increase was smaller than that observed in serum. In the CTL cows, the milking-induced PRL release above the premilking concentration was similar on d -1, 1, and 28 but was reduced during feed restriction. In the DOMP cows, the milking-induced PRL release was similar on d -1 and 1 but was reduced on d 28 and 34. Milk production was similar for both groups before the treatments started but was greater in the DOMP cows during the treatment period, at 2.9 ± 0.6 and 2.4 ± 0.6 kg/d greater during wk 3 and 4 of treatment, respectively. Milk production declined in both groups during feed restriction but remained higher in the DOMP cows. Milk production became similar again for both groups after the last injection. In addition, dry matter intake was increased by DOMP. These results support the hypothesis that PRL is galactopoietic in dairy cattle.

Effects of feed restriction and prolactin-release inhibition at drying-off on susceptibility to new intramammary infection in cows.

A cow's risk of acquiring a new intramammary infection during the dry period increases with milk production at drying-off. A method commonly used to reduce milk production is a drastic reduction in feed supply in the days that precede drying-off. Milk production can also be reduced by inhibiting the lactogenic signal driven by prolactin (PRL). This study aimed to compare the effects of these 2 drying-off procedures on milk production, metabolism, and susceptibility to intramammary infection in cows. A total of 21 Holstein cows in late lactation were assigned to 1 of 3 treatments based on milk yield, somatic cell count, and parity. The cows were fed a lactation diet until drying-off (control), only dry hay during the last 5 d before drying-off (DH), or the same diet as the control cows but with twice-daily i.m. injections of 4mg of quinagolide, a specific inhibitor of PRL release, from 5 d before drying-off until 13 d after (QN). On d 1 to 7 after the last milking, the cows were challenged by daily teat dipping in a solution containing Streptococcus agalactiae at 5×10(7) cfu/mL. Quinagolide induced a decrease in PRL concentration in blood on all the injection days. Blood PRL was also depressed in the hay-fed cows before drying-off. Both the QN and DH treatments induced a decrease in milk production, which at drying-off averaged 12.0, 10.0, and 21.7kg/d for the QN, DH, and control cows, respectively. The DH treatment decreased blood concentration of glucose and increased blood concentrations of ß-hydroxybutyrate and nonesterified fatty acids before drying-off. Somatic cell count at drying-off was greater in the milk of the QN cows than in that of the control cows but after drying-off was greater in the mammary secretions of the control cows than in those of the QN cows. The number of S. agalactiae colonies found in mammary secretions on d 8 and 14 after the last milking was lower for the QN cows than for the control cows. The percentage of S. agalactiae-infected quarters was also lower in the QN cows than in the control cows and on d 14 averaged 17.2, 33.7, and 57.5% in the QN, DH, and control cows, respectively. No differences between the DH and control groups were observed for either bacterial count or infection rate. In conclusion, this experiment shows that PRL-release inhibition could be an alternative for reducing milk production and improving resistance to intramammary infection at drying-off.

Effects of feed restriction and prolactin-release inhibition at drying off on metabolism and mammary gland involution in cows.

A cow's risk of acquiring a new intramammary infection during the dry period increases with milk production at drying off and decreases as mammary gland involution progresses. A method commonly used to reduce milk production is a drastic reduction in feed supply in the days that precede drying off. Milk production can also be reduced by inhibiting the lactogenic signal driven by prolactin (PRL). This study aimed to compare the effects of these 2drying-off procedures on metabolism, immunity, and mammary gland involution in cows. A total of 24Holstein cows in late lactation were assigned to 1 of 3treatments based on milk yield, somatic cell count, and parity. The cows were fed a lactation diet until drying off (control; n=8), only dry hay during the last 5d before drying off (DH; n=8), or the same lactation diet as the control cows but with twice-daily i.m. injections of 4mg of quinagolide, a specific inhibitor of PRL release, from 5d before drying off until 13d after (QN; n=8). Quinagolide induced a decrease in PRL concentration in blood and in milk and mammary secretions on all the injection days. Interestingly, PRL was also depressed in the blood and milk of the hay-fed cows before drying off. Both the QN and DH treatments induced a decrease in milk production, which averaged 17.9 and 10.1kg/d for the QN and DH cows, respectively, at drying off in comparison with 24.8kg/d for the control cows. Both BSA concentration and Na(+)-to-K(+) ratio increased faster in the mammary secretions of both the DH and QN cows than in those of the control cows, whereas citrate-to-lactoferrin ratio, another indicator of involution rate, decreased faster. The DH treatment decreased blood concentrations of glucose and most amino acids and increased blood concentrations of ß-hydroxybutyrate and nonesterified fatty acids. Quinagolide increased blood glucose but did not affect the other metabolites. The serum harvested on d-1 from the hay-fed cows reduced peripheral blood mononuclear cell proliferation and IL-4 production, whereas the serum from the quinagolide-treated cows had no effect. In conclusion, this experiment shows that PRL-release inhibition could be a new alternative for reducing milk production before drying off and for hastening mammary gland involution without disturbing the metabolism of the cow.

Effects of intramammary infusions of casein hydrolysate, ethylene glycol-bis(ß-aminoethyl ether)-N,N,N',N'-tetraacetic acid, and lactose at drying-off on mammary gland involution.

The transition from the lactation to the dry period in dairy cows is a period of high risk for acquiring new intramammary infections. This risk is reduced when involution of mammary glands is completed. Consequently, strategies that accelerate the involution process after drying-off could reduce the incidence of mastitis. The objective of this study was to assess the effect of 3 different treatments on mammary gland involution. Each quarter of 8 Holstein cows in late lactation was randomly assigned at drying-off to an intramammary infusion of casein hydrolysate (CNH; 70 mg), ethylene glycol-bis(ß-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA; 5.7 g), lactose (5.1g), or saline 0.9% (control) solutions. Milk samples were collected on the last 2 d before and 1, 3, 5, 7, 10, and 14 d after the last milking for determining concentrations of mammary gland involution markers. Lactoferrin, somatic cell counts (SCC), BSA, and Na(+) concentrations, as well as matrix metalloproteinase-2 and matrix metalloproteinase-9 activities gradually increased in mammary secretions during the first 2 wk following the last milking, whereas milk citrate and K(+) concentrations decreased. As involution advanced, the Na(+):K(+) ratio increased, whereas the citrate:lactoferrin ratio decreased. Compared with mammary secretions from control quarters, mammary secretions of quarters infused with CNH had higher SCC on d 1, 3, 5, and 7, and greater BSA concentrations on d 1, 3, and 5. Similarly, the CNH treatment induced a faster increase in lactoferrin concentrations, which were greater than in milk from control quarters on d 3, 5, and 7 after drying-off. Milk citrate concentrations were unaffected by CNH but the citrate:lactoferrin ratio was lower in CNH-treated quarters on d 3 and 5 than in control quarters. Moreover, CNH treatment hastened the increase in Na(+) concentration and in the Na(+):K(+) ratio on d 1. Infusion of CNH also led to an increase in proteolytic activities, with greater matrix metalloproteinase 9 activities on d 1 and 3. The EGTA infusion increased SCC above that of control quarters on d 1 and 3 but it had no effect on the other parameters. Lactose infusion had no effect on any of the involution markers. In this study, intramammary infusions of CNH were the most efficient treatment to accelerate mammary gland involution, suggesting a potential role of CNH as a local milk secretion inhibitor during milk stasis.

Effect of prolactin-release inhibition on milk production and mammary gland involution at drying-off in cows.

The end of each lactation is a challenging period for high-yielding cows as they are often dried off while still producing significant quantities of milk and, consequently, are highly susceptible to new intramammary infections. Once involution is complete, the mammary gland becomes much more resistant to infection. Therefore, it is critically important to develop strategies aimed at reducing milk production before drying-off and to accelerate mammary gland involution. This study assessed the effect of inhibition of the lactogenic signal driven by prolactin (PRL) on milk production and concentrations of involution markers in mammary secretions. Sixteen Holstein cows in late lactation were assigned to treatments based on milk yield, somatic cell count, and parity. Of those cows, 8 received twice-daily intramuscular injections (2 mg per injection) of quinagolide, a specific inhibitor of PRL release, from 4 d before drying-off to 3 d after (Quin). The other 8 cows received injections of the solvent (water, control). Blood and milk (mammary secretion) samples were collected on the last 5 d before and on d 1, 3, 5, 7, 10, and 14 after the last milking. Additionally, on the day preceding the first injection and on the following day, several blood samples were collected around milking time. Quinagolide reduced basal serum PRL concentrations on all injection days as well as PRL released in blood during milking. The PRL inhibitor decreased milk production before drying-off, which averaged, over the last 3 d of lactation, 19.3 and 15.5 kg/d for the control and Quin cows, respectively. Quinagolide had no significant effect on milk citrate:lactoferrin and Na:K ratios, which decreased and increased, respectively, during the first 2 wk of the dry period. Nevertheless, the increases in the number of somatic cells and bovine serum albumin concentration during early involution were greater and matrix metalloproteinase-2 activity tended to be greater in mammary secretions of the Quin cows compared with the control cows. This experiment shows that inhibition of PRL release decreases milk production of cows in late lactation. Changes in the composition of mammary secretions suggest that this approach also hastens mammary gland involution.

Whole intact rapeseeds or sunflower oil in high-forage or high-concentrate diets affects milk yield, milk composition, and mammary gene expression profile in goats.

This study aimed to ascertain the response of goat mammary metabolic pathways to concentrate and lipid feeding in relation to milk fatty acid (FA) composition and secretion. Sixteen midlactation multiparous goats received diets differing in forage-to-concentrate ratio [high forage (HF) 64:36, and low forage (LF) 43:57] supplemented or not with lipids [HF with 130 g/d of oil from whole intact rapeseeds (RS) and LF with 130 g/d of sunflower oil (SO)] in a 4 x 4 Latin square design. Milk yield, milk composition, FA profile, and FA secretion were measured, as well as the expression profiles of key genes in mammary metabolism and of 8,382 genes, using a bovine oligonucleotide microarray. After 3 wk of treatment, milk, lactose, and protein yields were lower with HF-RS than with the other diets, whereas treatment had no effect on milk protein content. Milk fat content was higher with the HF-RS and LF-SO diets than with the HF and LF diets, and SO supplementation increased milk fat yield compared with the LF diet. Decreasing the forage-to-concentrate ratio from 64:36 to 43:57 had a limited effect on goat milk FA concentrations and secretions. Supplementing the LF diet with SO changed almost all the FA concentrations, including decreases in medium-chain saturated FA and large increases in trans C18:1 and C18:2 isomers (particularly trans-11 C18:1 and cis-9, trans-11 conjugated linoleic acid), without significant changes in C18:0 and cis-9 C18:1, whereas supplementing the HF diet with RS led to a strong decrease in short- and medium-chain saturated FA and a very strong increase in C18:0 and cis-9 C18:1, without significant changes in trans C18:1 and conjugated linoleic acid. Despite the decreases in milk lactose and protein yields observed with HF-RS, and despite the decrease in milk medium-chain FA and the increase in C18 FA secretion with RS or SO supplementation, none of the dietary treatments had any effect on mammary mRNA expression of the key genes involved in lactose (e.g., alpha-lactalbumin), protein (e.g., beta-casein), and lipid metabolism (e.g., lipoprotein lipase) after 3 wk of treatment. In addition, transcriptome analysis did not provide evidence of treatments inducing significant changes in the expression of specific genes in the mammary gland. However, 2-way hierarchical clustering analysis highlighted different global mammary expression profiles between diets, showing that the gene expression profiles corresponding to the same diet were gathered by common groups of genes. This experiment suggests that after 3 wk of dietary treatment, other factors, such as substrate availability for mammary metabolism, could play an important role in contributing to milk FA responses to changes in diet composition in the goat.

Innovative dairy cow management to improve resistance to metabolic and infectious diseases during the transition period.

The incidence of metabolic and infectious diseases varies greatly during the lactation cycle. Most new cases of clinical mastitis appear at the beginning of lactation, and the incidence increases with the level of milk production. In addition to mastitis, many other infectious diseases become clinically apparent during the first 2weeks of lactation. During this time, cows are in a negative energy balance and must mobilize body reserves to balance the deficit between food energy intake and energy required for milk production. The relationships between energy deficit and metabolic diseases, such as ketosis and hepatic lipidosis, are well known. Furthermore, cows in energy deficit have a weakened immune system and are therefore more susceptible to infections. There is now good evidence that the increase in circulating non-esterified fatty acids impairs immune cell functions. Therefore, management approaches that reduce the negative energy balance and the increase in non-esterified fatty acids at the beginning of lactation are likely to improve resistance to infection. Improving the nutrient supply through periparturient nutritional management has been the subject of considerable research. However, another way to reduce the imbalance between nutrient supply and demand is to temporarily decrease the latter. In this review, we examine how management strategies such as conjugated linoleic acid feeding, prepartum milking, or limiting postpartum milk production could be used to reduce metabolic perturbations and immunosuppression during the transition period. At this stage, it appears that reducing the amount of milk harvested postpartum by means of partial milking in the first days after calving is the most promising approach to reduce metabolic stress and immunosuppression without compromising the productivity of high-yielding dairy cows.

Bronchial provocation testing of sodium iso-nonanoyl oxybenzene sulphonate.

1. Asthmatic symptoms have been reported in workers following occupational exposure to certain low molecular ratio chemicals. 2. A small number of workers involved in the initial manufacture of a new low temperature bleach activating chemical, sodium iso-nonanoyl oxybenzene sulphonate (SINOS) developed rashes, rhinitis and conjunctivitis following exposure to the compound. One worker also developed asthma. An investigative study was undertaken to examine the possible asthmatic effects of inhaling SINOS in naive non-asthmatic and asthmatic subjects handling the material in the laboratory setting. 3. Since SINOS has a similar chemical structure to aspirin, it was hypothesized that SINOS-associated asthma might be elicited by a mechanism similar to the mechanism associated with aspirin asthma. Therefore aspirin-sensitive asthmatics were also included in the study. 4. No adverse respiratory reactions were observed in the non-asthmatic subjects or in the aspirin and non-aspirin-sensitive patient volunteers following exposure to SINOS dust at atmospheric concentrations of up to 36.3 micrograms m-3. 5. Skin prick tests to increasing concentrations of SINOS were carried out in all subjects. No positive reactions were observed on any occasion. 6. This study indicates that SINOS does not elicit asthma via a mechanism similar to aspirin. Additionally, the study suggests that the addition of SINOS to washing powder will not cause significant respiratory reactions in consumers even if they are asthmatic or intolerant to aspirin.

[Generalized edema caused by licorice: a new syndrome. Apropos of 3 cases]. / Oedèmes généralisés induits par la réglisse: un nouveau syndrome. A propos de 3 observations.

Licorice abuse is a wellknown cause of high blood pressure, myopathy, and cardiac rhythm trouble. It should be considered as a cause of diffuse acute edema, as shown in the three following case-reports.

[Neurologic and psychiatric manifestations of primary hyperparathyroidism. Study of 5 cases]. / Manifestations neurologiques et psychiatriques de l'hyperparathyroïdie primitive. Etude de 5 cas.

Among 8 of the 20 records of primary hyperparathyroidism examined in search of neurological and/or psychiatric manifestations, the authors extracted 5 clinical cases reported here. These 5 cases were selected because these manifestations had been well evaluated before and after curative surgery of the disease. Among the manifestations reported, there was one corpus striatum syndrome, one cervical myelopathy and three cases of "chronic" psychiatric disorders of several years duration completely cured after parathyroidectomy. The authors underline that such psychiatric disorders should be taken into account in the discussion of treatment.

[Neoplastic hypercalcemia: prognostic factors of survival of patients; from 51 cases seen in internal medicine]. / Hypercalcémies néoplasiques: facteurs pronostiques pour la survie des patients; à partir de 51 cas observés en médecine interne.

In our Internal Medicine department, we conducted a retrospective study of prognostic factors in patients with malignant hypercalcaemia. The records of 51 patients who had both hypercalcaemia and a histologically proven cancer were analyzed; 42 had a solid tumour and 9 had a myeloma. In 61% of the patients cancer had been revealed by hypercalcaemia. The main warning signs were alteration of the general condition (68.6%), pain in the bones (54.9%) and polyuria with dehydration (58.8%). Osteolysis was observed in 75% of the cases. The overall median survival was 86 days. Patients with myeloma had a significantly longer survival than patients with other tumours (312 versus 60 days; p < 0.05). Patients who had received a causal treatment had a longer survival (176 versus 36 days, p < 0.001). In patients with solid tumours we found a negative correlation between survival and initial calcaemia, and a positive correlation between phosphoraemia, albuminaemia and survival. Multivariate analysis showed that the initial calcaemia level and the possibility of causal treatment were the two cardinal prognostic factors. Although the overall survival rate is mediocre, we believe that hospitalization of patients with malignant hypercalcaemia is justified for their better survival comfort and for the possibility of discovering a neoplasia that could benefit from an effective causal treatment, which is the principal factor of improved prognosis.

[Non-complicated Horton's disease: initial treatment with methylprednisolone 500 mg/day bolus for three days followed by 20 mg/day prednisone-equivalent. Evaluation of 15 patients]. / Maladie de Horton non compliquée: traitement initial par trois bolus de 500 mg de méthylprednisolone suivis de 20 mg/j d'équivalent-prednisone. Evaluation chez 15 patients.

PURPOSE: To assess the efficacy and tolerance of three methylprednisolone boluses (500 mg/d) followed by a standard dose of prednisolone, 20 mg/d, as the initial treatment of non-complicated giant-cell arteritis. METHOD: A retrospective study of 15 cases. RESULTS: Six men and nine women with a mean age of 70.9 years were treated and followed for 41.5 months. Initial mean ESR was 83 mm; mean C-reactive protein level was 94.6 mg/L. The boluses were well tolerated, excepted in one patient who developed acute psychosis. After initiating the oral treatment, two patients presented signs of clinical relapse during the first month, and were given higher doses of corticosteroids. At 1 month, 12 patients were asymptomatic, nine of whom had normalized ESR and CRP. Mean ESR was 23; mean CRP was 13 mg/L. At 3 months, the mean prednisone dose delivered was 18.2 mg/d. Mean ESR was 12 mm. The cumulative prednisone dose given during the first year was 5,349 (+/- 2,512) mg. In the 13 patients who necessitated no more than 20 mg/d prednisone, no sequelae of giant-cell arteritis, no fractures nor major treatment intolerance occurred-during the first 2 years of treatment. Treatment was stopped in eight patients after a mean duration of 48.6 months. CONCLUSION: Treatment with pulse methylprednisolone 500 mg/d for 3 days followed by 20 mg/d oral prednisone could be a valuable corticosteroid-sparing strategy in many patients with uncomplicated temporal arteritis.

[Oral histoplasmosis 34 years after return of Africa]. / Une histoplasmose du plancher buccal 34 ans après un retour d'Afrique.

INTRODUCTION: Histoplasmosis is a tropical fungal infection sharing many similarities with tuberculosis: the transmission by air dropplets, the usually asymptomatic primary-infection, the disseminated infection encountered among immunosuppressed patients and the granulomatous pathological lesions. In France, histoplasmosis is uncommon and may be misdiagnosed as tuberculosis. OBSERVATION: A 78 years old male patient presents with a raspberry-like lesion of the mouth causing difficulties to eat and weight loss of 14 kg. The diagnosis of tuberculosis is evoked because of the presence of a giant-cell granuloma in one of the biopsies. The histoplasmosis serology, requested because the patient stayed in Africa, is positive. Revisions of the pathology put into evidence the presence of spores in histiocytes confirming diagnosis of histoplasmosis. The treatment with itraconazole is effective. CONCLUSION: Histoplasmosis is a differential diagnosis of tuberculosis, especially in endemic regions. The histoplasmosis serology can be useful. The reference in diagnosis examinations keeps being the microscopic observation of spores and their mycological growth.