Minimum number of spermatozoa required for normal fertility after deep intrauterine insemination in non-sedated sows

A fibreoptic endoscope procedure for non-surgical deep intrauterine insemination in non-sedated sows has been reported. However, the endoscope is an expensive and fragile instrument, and is unsuitable for use under field conditions. The aim of this study was to determine the minimum number of spermatozoa required to maintain optimal fertility using a flexible catheter (1.8 m in length, 4 mm in diameter) for deep intrauterine insemination in 2-6 parity non-sedated sows. Crossbred sows were treated with eCG 24 h after weaning and with hCG 72 h later to induce oestrus. Deep intrauterine insemination was performed 36 h after hCG treatment in 117, 126, 60 and 69 sows with 15.0, 5.0, 2.5 or 1.0 x 10(7) spermatozoa in 10 ml, respectively. Weaned sows (n = 147) not treated with hormones and used for standard artificial insemination (AI) (two inseminations per oestrus with 3 x 10(9) spermatozoa in 100 ml) served as controls. The flexible catheter was passed successfully through the cervix into one uterine horn in 95.4% of the sows in an average of 3.7 +/- 0.09 min. Farrowing rates after deep intrauterine insemination with 15 or 5 x 10(7) spermatozoa did not differ from those of the control group (82.9, 76.2 and 83.0%, respectively), but a significant decrease (P < 0.001) was observed in sows inseminated with 2.5 or 1.0 x 10(7) spermatozoa (46.7 and 39.1%, respectively). In contrast, the number of spermatozoa inseminated did not affect prolificacy. Laparotomy revealed that the tip of the flexible catheter reached approximately the anterior third of the uterine horn. Although deep intrauterine insemination was performed in only one uterine horn, the percentages of embryos collected from the tip of both uterine horns 2 days after deep insemination were not significantly different. The results show that in comparison with standard AI, a 20-60-fold reduction in the number of spermatozoa inseminated and an 8-10-fold reduction in the dose volume can be achieved without decreasing fertility when semen is deposited non-surgically into the upper first third of one uterine horn.     

Generation of live rat offspring by intrauterine insemination with epididymal spermatozoa cryopreserved at -196 degrees C

This study reports the development of a reliable method for cryopreservation of rat epididymal spermatozoa and the production of live young by artificial insemination using these cryopreserved spermatozoa. The motility and membrane integrity of rat spermatozoa were investigated after spermatozoa had been subjected to physical stress and frozen with various concentrations of glycerol (0, 3 and 6%) either in the presence or absence of Equex Stem as cryoprotective agents. The ability of cryopreserved spermatozoa to generate normal offspring by intrauterine insemination was also evaluated. Rat spermatozoa that had been centrifuged at 700 g for 5 min showed a significant decrease in motility compared with non-centrifuged spermatozoa. In addition, after centrifugation three times the percentage of membrane-intact spermatozoa decreased to approximately 0%. The percentage of membrane-intact spermatozoa was significantly higher (P < 0.01) in semen samples that had been frozen in medium without glycerol than in samples frozen in medium with 3% glycerol. Although the addition of 0.7% Equex Stem to medium without glycerol or with 3% glycerol did not influence rates of sperm motility after freezing and thawing, the percentage of membrane-intact spermatozoa was improved by the presence of 0.7% Equex (P < 0.05). Therefore, rat spermatozoa were handled gently to avoid physical stress and were frozen in medium containing 23% egg yolk, 8% lactose monohydrate and 0.7% Equex Stem, at pH 7.4 adjusted with 10% Tris(hydroxymethyl)aminomethane solution. Thirteen female rats were inseminated into the oviductal end of both uterine horns with frozen-thawed spermatozoa. Forty-one normal live offspring were obtained from nine of the inseminated females. These results indicate that frozen-thawed rat spermatozoa can generate normal offspring. To our knowledge, this procedure is the first successful production of offspring using spermatozoa cryopreserved in liquid nitrogen.     

Effect of intrauterine insemination with spermatozoa or foreign protein on the mechanism of action of oestradiol in the rat oviduct

Previously, we showed that oestradiol accelerates oviductal egg transport through a non-genomic action involving oviductal protein phosphorylation in non-mated rats, and through a genomic action in mated rats. Thus, sensory stimulation, seminal fluid or sperm cells may be the source of signals that switch the mechanism of action of oestradiol in the oviduct to a genomic pathway. The present study examined the ability of spermatozoa to switch the mode of action of oestradiol in the absence of the sensory stimulation and seminal fluid provided by mating. Pro-oestrous rats were inseminated in each uterine horn with epididymal spermatozoa and 12 h later were injected subcutaneously with oestradiol and intrabursally with the mRNA synthesis inhibitor alpha-amanitin. The number of eggs in the oviduct, assessed 24 h later, showed that alpha-amanitin blocked the oestradiol-induced egg transport acceleration, indicating that the interaction of spermatozoa with the genital tract shifts the action of oestradiol from non-genomic to genomic. Other rats were inseminated with live or dead spermatozoa and then treated with the protein kinase inhibitor H-89, and oestradiol. Treatment with H-89 did not block the oestradiol-induced acceleration of egg transport in these rats, although dead spermatozoa did not enter the oviduct, indicating that the mere presence of spermatozoa in the uterus abrogated the non-genomic action of oestradiol in the oviduct. Treatment with H-89 also failed to prevent the acceleration of oviductal egg transport induced by oestradiol in rats inseminated with hamster spermatozoa or with BSA, whereas in rats inseminated with their own serum (autologous proteins), H-89 was able to prevent the effect of oestradiol. This finding reveals that the effect of insemination on the mode of action of oestradiol is neither species-nor sperm-specific and it is produced by foreign organic material. It can be concluded that the presence of spermatozoa or foreign protein in the uterus is one of the components of mating that is capable of switching the action of oestradiol in the oviduct from a non-genomic to a genomic mode.     

Transport and fate of spermatozoa after insemination of cattle

Sperm capable of fertilizing ova reach the isthmus of cows about 8 h after mating and remain in the caudal 2 cm of the isthmus until ovulation. Then small numbers of sperm move to the site of fertilization at the junction of the isthmus and ampulla. Within a few hours after deposition of semen in the uterine body, most sperm have drained to the exterior in cervical mucus. By 12 to 24 h after insemination, only a few percent of the sperm remain in the reproductive tract, and most of these are in the vagina. Contractions of the reproductive tract appear to be the primary mechanism of sperm transport. Flagellation of sperm is probably required for sperm to enter the folds of the cervix, and flagellation may be helpful or essential for sperm to pass through the uterotubal junction, move from the isthmus to the ampulla, and penetrate ova. High proportions of sperm undergo the acrosome reaction only in the ampulla on the side of ovulation and only after ovulation. The fertilization rate in cattle can be improved by use of semen from high fertility bulls and perhaps by timing insemination with semen from lower fertility bulls after the end of estrus.     

Neutrophil recruitment and phagocytosis of boar spermatozoa after artificial insemination of sows,and the effects of inseminate volume,sperm dose and specific additives in the extender

In this study the recruitment of leucocytes and phagocytosis of spermatozoa after artificial insemination of multiparous sows was investigated. In Expt 1, groups of sows received either no inseminate (n = 6) or inseminates with various concentrations of spermatozoa and seminal plasma or different inseminate volumes (n = 9 per group). In Expt 2, groups of sows received inseminates containing no addition, caffeine + CaCl(2), or excess EDTA (n = 6 per group). Leucocytes and spermatozoa were counted in the collected backflow from the vulva, and in the PBS flushings of the genital tract of sows killed at 4 h after insemination. Tissue homogenates were checked for remaining spermatozoa. Leucocyte recruitment did not depend on the presence of seminal plasma or spermatozoa. In the control groups about 43% of the inseminated spermatozoa were found in the backflow and 5% in the genital tract. Many spermatozoa could be recognized inside polymorphonuclear leucocytes. With an inseminate volume of 20 ml instead of 80 ml, fewer spermatozoa were found in the backflow and more (non-phagocytosed) spermatozoa were recovered in the uterus (P < or = 0.05). With a sperm dose of 0.24 x 10(9) instead of 2.4 x 10(9), a higher percentage of the inseminated spermatozoa was recovered in the oviducts (P < or = 0.05). The use of caffeine + CaCl(2) resulted in lower recruitment of leucocytes (P < or = 0.05) and a higher number of non-phagocytosed spermatozoa in the uterus (P < or = 0.01) compared with controls. The numbers of spermatozoa in the oviducts were not different. Insemination with excess EDTA had no positive effects on the number of spermatozoa in the genital tract.     

Hot topic: Successful fixed-time insemination within 21 d after first insemination by combining chemical pregnancy diagnosis on d 18 with a rapid resynchronization program

Cattle that are not pregnant to first fixed-time artificial insemination (TAI) may be resynchronized for a second TAI if they are found nonpregnant at pregnancy diagnosis. The specific interval between first and second TAI ranges from 4 to 8 wk. The selected interval depends on the available method of pregnancy diagnosis and the efficiency of the resynchronization program. The objective of this experiment was to evaluate a pregnancy diagnosis and resynchronization system that achieved a 21-d interval between TAI. This 21-d interval approximates the natural return-to-service interval. It also enables resynchronization to be implemented within the same estrous cycle in which cattle are first inseminated. Holstein heifers were randomly assigned to a 21-d resynchronization program (21d_resynch; n = 40) or a control group in which estrus was observed for the purpose of re-insemination (control; n = 29). The 21d_resynch heifers were diagnosed for pregnancy on d 18 after a TAI (d 0) by using predetermined cut-off values for 2′-5′ oligoadenylate synthetase 1 (Oas1) gene expression in leukocytes and plasma progesterone concentration. Heifers that were not pregnant to first TAI had greater expression of Oas1 at the time of PGF_2α (d −3) than pregnant heifers, but this relationship was reversed on d 18 after TAI: the heifers that were pregnant to first TAI had almost 5-fold greater expression of Oas1 compared with nonpregnant heifers. Nonpregnant heifers in the 21d_resynch group were injected with a luteolytic dose of PGF_2α on d 19 and were injected with GnRH on d 21 and submitted to TAI. The pregnancy per AI after first insemination was similar for 21d_resynch (50.0%; pregnancy diagnosis on d 18) and control (51.7%; pregnancy diagnosis on d 27). Likewise, no difference was detected in second insemination pregnancy per AI for 21d_resynch (36.8%; nonpregnant heifers TAI on d 21) and control (35.7%; nonpregnant heifers inseminated at return to estrus or after nonpregnant diagnosis on d 27). The interval between first and second insemination was shorter for 21d_resynch compared with control (21.0 ± 0 and 27.5 ± 2.1 d). The conclusion is that a TAI resynchronization can be programmed within 21 d of previous TAI when a d 18 pregnancy test and a rapid resynchronization are used.     

Estrous cycle characterisation and artificial insemination using frozen-thawed spermatozoa in the bottlenose dolphin (Tursiops truncatus)

The reproductive endocrinology of the bottlenose dolphin, Tursiops truncatus, was characterized to facilitate the development of artificial insemination using cryopreserved spermatozoa. Specific objectives were: (i) to determine the excretory dynamics of urinary luteinizing hormone (LH) and ovarian steroid metabolites during the estrous cycle; (ii) to evaluate the effect of an exogenously administered synthetic progesterone analog (altrenogest) on reproductive hormone excretion; (iii) to correlate follicular growth and ovulation (as determined by transabdominal ultrasound) to urinary LH and ovarian steroid metabolites; (iv) examine the in vivo fertilisation capacity of cryopreserved semen, and (v) to develop an intrauterine insemination technique. Based on urinary endocrine monitoring of natural estrous cycles (2 consecutive cycles) and nine post altrenogest cycles in ten females, estrous cycles were found to be 36 days long and comprised of an 8 day and 19 day follicular and luteal phase, respectively. Peak estrogen conjugates (EC; 5.4+/-3.8 ng/mg creatinine (Cr)) occurred 8 h prior to the LH surge (70.9+/-115.7 ng/mg Cr). The time of ovulation, as determined by ultrasonography, occurred 32.1+/-8.9 h and 24.3+/-7.0 h after the onset of the LH surge and LH peak, respectively. Mean preovulatory follicular diameter and circumference were 2.1+/-0.5 cm and 6.5+/-1.5 cm, respectively. Of the 27 estrous synchronisation attempts, 13 resulted in an ovulatory cycle, with ovulation occurring 21 days post-altrenogest treatment. Intrauterine (4 of 5) and intracornual (1 of 3) inseminations conducted across eight estrous cycles resulted in five pregnancies (63%), one pregnancy resulted from the use of liquid stored semen, whereas four were achieved using cryopreserved semen. These data provide new information on female bottlenose dolphin reproductive physiology, and demonstrate that the combination of endocrine monitoring and serial ultrasonography contributed to successful AI using liquid-stored and cryopreserved semen.     

Effects of intrauterine infusion of seminal plasma at artificial insemination on fertility of lactating Holstein cows

An inflammatory response is induced in the reproductive tract by deposition of semen during natural mating. This response might facilitate establishment and maintenance of pregnancy and alter the phenotype of the offspring by modifying the microenvironment of the reproductive tract. Here, we hypothesized that intrauterine infusion of 0.5 mL of seminal plasma at the time of artificial insemination (AI) in first-service lactating Holstein cows will improve pregnancy success after insemination. Cows were inseminated (511 primiparous cows inseminated with X-sorted semen, 554 multiparous cows inseminated with X-sorted semen, and 627 multiparous cows inseminated with conventional semen) using the Double-Ovsynch protocol. Cows were randomly assigned to receive intrauterine infusion of either 0.5 mL of seminal plasma or saline immediately after AI. There was no overall effect of seminal plasma infusion on the percentage of inseminated cows diagnosed pregnant at d 32 or 60 after AI, pregnancy loss, or percent of inseminated cows calving. If cows were inseminated with conventional semen, seminal plasma reduced pregnancies at d 32 and tended to reduce calvings. There was no effect of seminal plasma if cows were inseminated with X-sorted semen. Seminal plasma infusion increased the birth weight of heifer calves born using X-sorted semen but not conventional semen. These results do not support a beneficial effect of seminal plasma on pregnancy success after AI, but exposure to seminal plasma may program fetal development to affect phenotype at birth.     

Timed artificial insemination with estradiol cypionate or insemination at estrus in high-producing dairy cows

A total of 799 Holstein cows from 3 herds were randomly assigned at 37 +/- 3 d in milk (DIM) to timed artificial insemination (AI) or insemination at detected estrus. Cows were presynchronized with injections of PGF(2alpha) at 37 and 51 DIM. At 65 DIM, cows received an injection of GnRH, followed 7 d later by PGF(2alpha). Cows in the estrus-detected group were inseminated after being observed in estrus during the 7 d after the last PGF(2alpha). Cows in the timed AI group received an injection of 1 mg of estradiol cypionate (ECP) 24 h after the last PGF(2alpha). If detected in estrus or=1 ng/mL; L = <1 ng/mL), resulting in 8 combinations (LLL, LHL, LLH, LHH, HHH, HHL, HLH, and HLL). Conception rates and pregnancy rates were higher for cows in the timed AI group than in the estrus-detected group at 30, 44, and 58 d (e.g., at 58 d, pregnancy rates were 42.2% for multiparous cows or 34.4% for primiparous cows in the group receiving ECP and timed AI compared with only 20.8 or 18.8% for respective parity subgroups for the treatment group inseminated only at detected estrus). Pregnancy losses were 11.5% from 30 to 58 d and did not differ between treatments. Cyclic cows within both treatments had higher estrous responses, conception rates, and pregnancy rates. Cows that responded to presynchronization and to luteolysis (HHL) had the highest conception and pregnancy rates, followed by cows classified as LHL. Use of 1 mg of ECP to induce ovulation as part of a synchrony regimen improved reproduction at first postpartum insemination in dairy cows.     

Some aspects of the statistics of small numbers of triangular taste tests

Summary. The binomial distribution of random selections of odd samples in small numbers of triangular taste tests has been applied to estimate: (a) scores establishing a difference between samples, (b) the extent of true discrimination shown by any significant score, (c) the fixed-panel size needed to establish difference or similarity, and (d) decisive scores in a sequential test scheme. It is shown that conclusions may be reached with fewer tests than the published literature suggests, thereby affording a saving of tasting effort.     

Hyperketolactia occurrence before or after artificial insemination is associated with a decreased pregnancy per artificial insemination in dairy cows

The reproductive parameters of dairy cattle have continuously declined worldwide over the last 50 years. Nutritional imbalances are identified as risk factors for this decrease of reproductive performance. The present paper aims to quantify the decrease in the pregnancy per artificial insemination (P/AI) in the case of high milk ketones before and after AI. A total of 388,731 test-day from the Brittany Milk Recording Program in France from 226,429 cow-lactations were provided for this trial. For each test-day, information about lactation characteristics, date of AI, date of the following calving, and acetone and β-hydroxybutyrate (BHB) values were included. Ketones were predicted by Fourier transform mid-infrared spectroscopy using MilkoScan Foss analyzers (Foss, Hillerød, Denmark). Many thresholds were evaluated to define cows with hyperketolactia. Hyperketolactia statuses were then categorized into 1 of 4 possible classes according to the milk ketone dynamics for each AI and each threshold of acetone or BHB values (low-low, high-low, low-high, and high-high) within 20 d before and after AI. Similarly, the dynamics of udder health were characterized by changes in somatic cell counts measured at the same test day as ketone bodies. A logistic regression with a Poisson correction was performed to explain the relationship of P/AI with milk ketones and somatic cell count dynamics. Predicted acetone and BHB ranged from −0.51 to 4.92 mM (mean = 0.08 mM, SD = 0.10 mM) and −0.62 to 5.85 mM (mean = 0.07 mM, SD = 0.1 mM), respectively. Hyperketolactia defined by high acetone levels before AI was not associated with decreased P/AI, but high acetone levels after AI were associated with a >10% reduction in P/AI for all thresholds >0.10 mM. Hyperketolactia, defined by high BHB values before, after, or before and after AI, was associated with a 6 to 14% reduction in P/AI compared with cows with low BHB values. These associations are lower than those reported in previous trials in which blood ketones were used. High ketones in advanced lactation are likely to be the result of various primary disorders (secondary ketosis). Because the present work demonstrated that this situation is considered a risk factor for deteriorated reproductive performance, we suggest that high ketones in early and advanced lactation should be of interest to farm advisors.     

Performance of automated activity monitoring systems used in combination with timed artificial insemination compared to timed artificial insemination only in early lactation in dairy cows

Identifying cows in estrus remains a challenge on dairy cattle farms, and tools and technologies have been developed and used to complement or replace visual detection of estrus. Automated activity monitoring (AAM) systems and timed artificial insemination (TAI) are technologies available to dairy farmers, but many factors can influence their relative performance. The objective of the present study was to compare reproductive performance of cows managed with an AAM system combined with TAI, or with a TAI program (Double Ovsynch) for insemination before 88 DIM. From April 2014 to April 2015, 998 cows from 2 herds were randomly assigned either to be inseminated at 85 ± 3 DIM exclusively using the Double Ovsynch protocol for TAI, or to be inseminated based on estrus detection by AAM without hormonal intervention between 50 and 75 DIM; if no alarm was detected by 75 DIM, cows were inseminated following the single Ovsynch protocol (AAM + Ovsynch). The herds used different AAM systems. Parity, hyperketonemia at wk 1 and 2 postpartum (PP), purulent vaginal discharge at wk 5 PP, body condition score at wk 7 PP, and anovulation to wk 9 PP were recorded. These health indicators did not significantly differ between treatments, but did between herds. The effect of treatment on pregnancy at first insemination and by 88 DIM were assessed using logistic regression models. Time to pregnancy was assessed using survival analysis. Results are reported from intention-to-treat analyses. Treatment did not affect pregnancy at first insemination or pregnancy by 88 DIM, but we found significant interactions between treatment and herd for both outcomes. In herd 2, marginal mean pregnancy at first AI was greater with Double Ovsynch (38%) than AAM + Ovsynch (31%), but no difference was observed in herd 1 (Double Ovsynch = 31%; AAM + Ovsynch = 34%). By 88 DIM, a smaller proportion of cows in herd 1 were pregnant in Double Ovsynch (31%) than AAM + Ovsynch (49%), but there was no difference in herd 2 (Double Ovsynch = 38%; AAM + Ovsynch = 38%). We observed a treatment by herd interaction for median (95% confidence interval) time to pregnancy, which were, in herd 1, 110 (106 to 129) and 98 (88 to 113) d, and, in herd 2, 126 (113 to 139) and 116 (105 to 131) d for the Double Ovsynch and AAM + Ovsynch treatments, respectively. The relative performance of AAM-based reproductive management compared with TAI only is likely influenced by herd-specific variables, in particular related to insemination rate when estrus detection is employed.     

Timing of insemination in dairy heifers.

Data representing 676 AI of 372 virgin dairy heifers during 1984 to 1987 at the University of Illinois were analyzed by analysis of variance to determine the effect of timing of AI on conception rate. Standing to be mounted, bawling, and attempting to mount were the three criteria used for determining the presence of estrus. More than 90% of the observations were conducted by one herdsperson between 0700 and 1600 h daily. Heifers were artificially inseminated with frozen semen from 58 bulls by one of two technicians. Conception rate on first AI was 56%, determined by return to estrus or 33- to 50-d rectal palpation for pregnancy. Conception rate to second and later AI was near 40%. Eighty-six percent of the heifers conceived during the trial. The average interval between first observation of standing estrus and AI was 10.5 h; the range was 15 min to 33 h. The model discussed contained six variables: time interval between first observation of standing estrus and AI (group), breed, sire within breed, year bred, season bred, and inseminator. The analysis of variance of conception to first AI showed timing of AI and service sire as the two most significant effects. Conception to first AI was worst when AI occurred 13.5 to 33 h after first observation of estrus. No factors considered were significant sources of variation for second or later AI.     

Increasing the length of an estradiol with progesterone timed artificial insemination protocol with 2 controlled internal drug release devices improves pregnancy per artificial insemination in lactating dairy cows

The objective of this study was to compare the effects of different lengths of ovulation synchronization protocols using 2 controlled internal drug release (CIDR) devices on ovarian dynamics and pregnancy outcomes in lactating dairy cows. Lactating Holstein cows (n = 1,979) were randomly assigned to receive timed artificial insemination (TAI; d 0) following 1 of 2 treatments: (1) 9-d protocol (n = 988; 9D) with 2 intravaginal devices containing 1.9 g of progesterone (CIDR) and 2.0 mg of estradiol benzoate on day ?11; 25 mg (i.m.) of dinoprost tromethamine (PG) and withdrawal of 1 CIDR on d ?4; 1.0 mg (i.m.) of estradiol cypionate, second CIDR withdrawal, and PG on d ?2; and TAI on d 0 and (2) 10-d protocol (n = 991; 10D) with 2 CIDR and 2.0 mg of estradiol benzoate on d ?12; 25 mg of PG and withdrawal of 1 CIDR on d ?4; 1.0 mg of estradiol, second CIDR withdrawal, and PG on d ?2; and TAI on d 0. There was no effect of protocol on estrus detection, whereas a greater percentage of cows from 10D had ovulated close to TAI [no corpus luteum (CL) at AI and a CL at d 7] versus cows assigned to 9D protocol. A protocol × heat stress (average cow temperature ≥39.1°C on day of AI and d 7) interaction was observed in a manner that pregnancy per AI (P/AI) was greater in non-heat-stressed 10D versus 9D cows, whereas P/AI did not differ when cows were under heat stress. Furthermore, 10D protocol did not increase P/AI when all cows that received AI were included in the analysis or in cows that ovulated near TAI. However, animals assigned to 9D without any event of heat stress had a reduced P/AI when compared with cows assigned to 10D without heat stress. A protocol × CL presence at the beginning of the protocol interaction was observed and cows with a CL at the beginning of the protocol had a greater P/AI in 10D versus 9D; meanwhile, in cows without a CL, no differences on P/AI were observed. The protocol × CL presence at the beginning of the protocol interaction on P/AI was also observed for cows that ovulated near TAI. A greater percentage of cows assigned to 9D had follicles of medium size (13–15.9 mm), and greater percentage of cows assigned to 10D had larger follicles (>16 mm). Increasing the length of an estradiol with progesterone–based ovulation synchronization protocol (10D vs. 9D) increased the proportion of cows with larger follicles (>16 mm) and increased P/AI in cows without heat stress and in cows with a CL at beginning of the protocol. Moreover, the 10D protocol increased the proportion of cows with ovulation near TAI, demonstrating the effectiveness of this protocol in improving the reproductive performance of lactating Holstein cows.     

Low prevalence of Listeria monocytogenes in cull sows and pork

The goal of this study was to determine the prevalence of Listeria monocytogenes in sows slaughtered at a single Midwestern plant on two occasions (trial 1, n = 179 sows; trial 2, n = 160 sows). Fecal samples collected antemortem (trial 1) as well as animal tissues, and carcass swabs collected at the abattoir (trials 1 and 2) were analyzed. Eight isolates of L. monocytogenes were recovered from five samples that represented 0.18% of the total samples (n = 2,775). In trial 1, L. monocytogenes was detected in a tonsil sample (0.6%; 1 positive of 181 tonsils), in a carcass (0.6%; 1 positive of 179 carcasses), which was sampled prior to the organic rinse, and in two chopped meat block samples (1.2%; 2 positive of 165 samples). In trial 2, L. monocytogenes was only detected in a single chopped meat block sample (0.15%; 1 positive of 688 total samples). These data indicate the low prevalence of L. monocytogenes in the cull sow.     

A simple procedure for the detection of small numbers of enterotoxigenic Clostridium perfringens in frozen meat and cod paste

By use of commercially available pouches and dehydrated modified Handford medium, a simple method for the selective detection of small numbers of Clostridium perfringens spores in frozen meat and cod paste has been developed. By a direct method, 1 × 10²/g or more spores were detectable and after enrichment culture, the sensitivity of the direct method was increased by 10 to 100 times. One hundred and thirty of 319 samples (40.8%) of frozen meat and cod fish paste proved positive for presumptive C. perfringens by the enrichment method. The incidences of presumptive C. perfringens in frozen mutton (50.0%), rabbit meat (65.0%), cod paste (50.0%) and horse meat (42.4%) were relatively higher than those in frozen beef (25.0%) and pork (23.8%). By the direct method, the incidence decreased to 3.1% (10/319). Two enterotoxigenic strains were found among 100 verified C. perfringens isolates from 58 samples.     

Changes in milk urea around insemination are negatively associated with conception success in dairy cows

Dietary protein levels are a risk factor for poor reproductive performance. Conception is particularly impaired in cases of high blood or milk urea. The objective of this study was to investigate the association between conception and low milk urea or changes in milk urea around artificial insemination (AI). Data were obtained from the French Milk Control Program for a 4-yr period (2009–2012). Milk urea values between 250 and 450 mg/kg (4.3 and 7.7 mM) were considered intermediate (I), and values ≤150 mg/kg (2.6 mM) were considered low (L). Milk urea values before and after each AI were allocated into 4 classes representing the dynamics of milk urea (before-after; I-I, I-L, L-I, and L-L). Subclinical ketosis was defined using milk fat and protein contents before AI as proxies. A logistic regression with a Poisson correction and herd as a random variable was then performed on data from Holstein or all breeds of cows. The success of conception was decreased [relative risk (95% confidence interval) = 0.96 (0.94–0.99)] in low-urea cows compared with intermediate-urea cows after AI; no significant association was found for urea levels before AI. When combining data on urea before and after AI, I-L urea cows exhibited a 5 to 9% decrease in conception compared with I-I urea cows, and L-I urea cows showed no difference in conception success compared with I-I urea cows. A decreased conception success for L-L urea cows compared with I-I urea cows was observed for the analysis with cows of all breeds. This work revealed that a decrease in urea from intermediate (before AI) to low (after AI) is a risk factor for conception failure. Surveys of variation in milk urea in dairy cows close to breeding are highly recommended.