Reproductive performance of lactating dairy cows with elevated milk ß-hydroxybutyrate levels during first 6 weeks of lactation.

Although there is evidence that ketosis negatively affects fertility, the effect of late and early ketosis on the reproductive performance of lactating cows has not been systematically investigated. The aim of this study was to evaluate the association between time and amplitude of elevated milk BHB (EMB) occurring within 42 d in milk (DIM) and subsequent reproductive performance of lactating Holstein cows. The dairy herd information data of 30,413 cows with 2 test-day milk BHB recordings during early lactation periods 1 and 2 (5-14 and 15-42 DIM, respectively) assessed as negative (<0.15 mmol/L), suspect (0.15-0.19 mmol/L), or positive (≥0.2 mmol/L) for EMB were used in this study. Based on the time and amplitude of milk BHB, cows were grouped into 7 groups: (1) healthy cows negative in both periods 1 and 2 were classified as NEG; (2) suspect in period 1 and negative in period 2: EARLY_SUSP; (3) suspect in period 1 and suspect/positive in period 2: EARLY_SUSP_Pro; (4) positive in period 1 and negative in period 2: EARLY_POS; (5) positive in period 1 and suspect/positive in period 2: EARLY_POS_Pro; (6) negative in period 1 and suspect in period 2: LATE_SUSP; and (7) negative in period 1 and positive in period 2: LATE_POS. The overall prevalence of EMB within 42 DIM was 27.4%, with the highest prevalence being EARLY_SUSP (10.49%). Cows in EARLY_POS and EARLY_POS_Pro, but not other EMB categories, had a longer interval from calving to first service compared with NEG cows. For the reproductive parameters, first service to conception interval, days open and calving interval, cows in all EMB groups except EARLY_SUSP had longer intervals compared with NEG cows. These data indicate that there is a negative association between EMB within 42 d and reproductive performance after the voluntary waiting period. The intriguing findings of this study are the unaltered reproductive performance of EARLY_SUSP cows, and the negative association between late EMB and reproductive performance. Hence, monitoring and prevention of ketosis during the first 6 wk of lactation is necessary to optimize reproductive performance of lactating dairy cows.

Effects of Herbicides and the Chytrid Fungus Batrachochytrium dendrobatidis on the growth, development and survival of Larval American Toads (Anaxyrus americanus).

Pesticides and pathogens adversely affect amphibian health, but their interactive effects are not well known. We assessed independent and combined effects of two agricultural herbicides and the fungal pathogen Batrachochytrium dendrobatidis (Bd) on the growth, development and survival of larval American toads (Anaxyrus americanus). Wild-caught tadpoles were exposed to four concentrations of atrazine (0.18, 1.8, 18.0, 180 µg/L) or glyphosate (7, 70, 700, 7000 µg a.e./L), respectively contained in Aatrex® Liquid 480 (Syngenta) or Vision® Silviculture Herbicide (Monsanto) for 14 days, followed by two doses of Bd. At day 14, atrazine had not affected survival, but it non-monotonically affected growth. Exposure to the highest concentration of glyphosate caused 100% mortality within 4 days, while lower doses had an increasing monotonic effect on growth. At day 65, tadpole survival was unaffected by atrazine and the lower doses of glyphosate. Neither herbicide demonstrated an interaction effect with Bd on survival, but exposure to Bd increased survival among both herbicide-exposed and herbicide-control tadpoles. At day 60, tadpoles exposed to the highest concentration of atrazine remained smaller than controls, indicating longer-term effects of atrazine on growth, but effects of glyphosate on growth disappeared. Growth was unaffected by any herbicide-fungal interaction but was positively affected by exposure to Bd following exposure to atrazine. Atrazine exhibited a slowing and non-monotonic effect on Gosner developmental stage, while exposure to Bd tended to speed up development and act antagonistically toward the observed effect of atrazine. Overall, atrazine, glyphosate and Bd all showed a potential to modulate larval toad growth and development.

Comparison of cattle derived from in vitro fertilization, multiple ovulation embryo transfer, and artificial insemination for milk production and fertility traits.

The use of assisted-reproduction technologies such as in vitro fertilization (IVF) is increasing, particularly in dairy cattle. The question of consequences in later life has not yet been directly addressed by studies on large animal populations. Studies on rodents and early data from humans and cattle suggest that in vitro manipulation of gametes and embryos could result in long-term alteration of metabolism, growth, and fertility. Our goal was to better describe these presumed consequences in the population of dairy cows produced by IVF in Québec (Canada) and to compare them to animals conceived by artificial insemination (AI) or multiple ovulation embryo transfer (MOET). To do so, we leveraged a large phenotypic database (2.5 million animals and 4.5 million lactations) from milk records in Québec aggregated by Lactanet (Sainte-Anne-de-Bellevue, QC, Canada) and spanning 2012 to 2019. We identified 304,163, 12,993, and 732 cows conceived by AI, MOET, and IVF, respectively, for a total of 317,888 Holstein animals from which we retrieved information for 576,448, 24,192, and 1,299 lactations (total = 601,939), respectively. Genetic energy-corrected milk yield (GECM) and Lifetime Performance Index (LPI) of the parents of cows were used to normalize for genetic potential across animals. When compared with the general Holstein population, MOET and IVF cows outperformed AI cows. However, when comparing those same MOET and IVF cows with only herdmates and accounting for their higher GECM in the models, we found no statistical difference between the conception methods for milk production across the first 3 lactations. We also found that the rate of Lifetime Performance Index improvement of the IVF population during the 2012 to 2019 period was less than the rate observed in the AI population. Fertility analysis revealed that MOET and IVF cows also scored 1 point lower than their parents on the daughter fertility index and had a longer interval from first service to conception, with an average of 35.52 d compared with 32.45 for MOET and 31.87 for AI animals. These results highlight the challenges of elite genetic improvement while attesting to the progress the industry has made in minimizing epigenetic disturbance during embryo production. Nonetheless, additional work is required to ensure that IVF animals can maintain their performance and fertility potential.

Molecular characterization of persistent subclinical mastitis-causing Staphylococcus aureus from dairy farms.

The study aimed to evaluate the genetic diversity of Staphylococcus aureus causing subclinical mastitis (SM) isolated from dairy cows and to assess the effect of the infection status (transient vs. persistent) on the milk and component yield. A total of six dairy farms in São Paulo state were used for the selection of cows with SM caused by S. aureus. S. aureus strains (n = 56) obtained from three biweekly aseptic mammary quarter milk samplings (n = 1140 from 95 cows) were subjected to MALDI-TOF MS analysis for species confirmation and further PFGE analysis. Intramammary infections (IMI) caused by S. aureus were categorized as transient (T: when only one out of 3 milk samplings had positive isolation of any pulsotype) or persistent (P: when two (P2) or three (P3) milk samplings had positive isolation of identical pulsotype over the consecutive episodes of SM. The SmaI macrorestriction fragment profiles of 56 S. aureus isolates showed a dominant S. aureus clonal pattern (PFGE type A; n = 50; 89.3%) within and among the herds. The SM-causing S. aureus represented a reduction of quarter milk yield of 26.2% in transient and 54.8% in persistent cases as well as a reduction of total solid yield of 38.1% and 49.4%, respectively, when compared with the healthy control quarters. Overall, the greater chance of S. aureus to be persistent is when a dominant clonal pattern is present in the herd which consequently may be associated with the cause of accentuated milk loss.

Gestational and health outcomes of dairy cows conceived by assisted reproductive technologies compared to artificial insemination.

Herd gestation and health management are key aspects of effective dairy farm operations and animal welfare improvement. Unfortunately, very little is known about the developmental divergences induced by assisted reproduction technologies (ART) and their consequences once the animal is mature. Indeed, the gestational and health outcomes of this subset of the Holstein population is yet to be characterized. In this study, the intergenerational impacts of ART conception were assessed by looking at the gestation and health outcomes of a large cohort of cows (n = 284,813) for which the conception methods were known. Our results showed that cows conceived by multiple ovulation embryo transfer (MOET) and in vitro fertilisation (IVF) displayed longer gestations: +0.37 ± 0.079 and +0.65 ± 0.21 day compared to cows conceived by artificial insemination (AI). Surprisingly, animals conceived by all methods experienced a similar 1-day decline in average gestation length from 2012 to 2019. Cows conceived by IVF were not more likely to experience stillbirths but were affected by common diseases such as ovarian cysts, mastitis, and uterine diseases in different proportions compared to cows conceived by other methods. This study provides new and unique information on ART animals regarding perinatal mortality and general health outcomes.

The effect of new and ancestral inbreeding on milk production traits in Iranian Holstein cattle.

Inbreeding depression, the reduction of fitness and performance, is due to an increase in the mating of related individuals. Based on the purge hypothesis, inbreeding and breeding over generations reduce the effect of deleterious alleles responsible for inbreeding depression. Thus, recent inbreeding is assumed to be more harmful than ancestral inbreeding. This study aimed at evaluating the effects of new and ancestral inbreeding on milk, fat and protein production in Iranian Holstein cattle. The secondary objective was to examine the changes in predicted breeding values when the inbreeding effect was included in the model's analysis. To this end, inbreeding coefficients were calculated using the pedigree of 2,394,517 Holstein cattle to achieve these goals. In addition, 419,132 records of milk, fat and protein yields of first parity cows were collected to assess inbreeding depression and breeding values. The average inbreeding coefficients were 0.83% and 1.68% for the whole population and the inbred animals, respectively. A 1% increase in classical pedigree-based inbreeding coefficient was associated with a decrease of 11.99 kg in milk, 0.39 kg in fat and 0.29 kg in protein. The effect of ancestral inbreeding was more detrimental to performance traits than the effect of new inbreeding. This result contradicted the hypothesis of purging. By including the inbreeding coefficient in the model, the rank of animals remained unchanged, but the average predicted breeding values increased. In general, inbreeding depression was observed in Iranian Holstein cows; however, no evidence of purging was observed. The average of inbreeding coefficients was not high in this population, although accounting for inbreeding coefficients in the analytical model did significantly increase the predicted breeding values. It is recommended that the analytical model incorporate the inbreeding coefficient to improve the accuracy of genetic evaluation. In future studies, inbreeding depression should be assessed using genomic data for performance and reproduction traits.

Incidence and Treatments of Bovine Mastitis and Other Diseases on 37 Dairy Farms in Wisconsin.

The aim of this research was to describe the incidence and treatments of mastitis and other common bovine diseases using one year of retrospective observational data (n = 50,329 cow-lactations) obtained from herd management software of 37 large dairy farms in Wisconsin. Incidence rate (IR) was defined as the number of first cases of each disease divided by the number of lactations per farm. Clinical mastitis (CM) remains the most diagnosed disease of dairy cows. Across all herds, the mean IR (cases per 100 cow-lactations) was 24.4 for clinical mastitis, 14.5 for foot disorders (FD), 11.2 for metritis (ME), 8.6 for ketosis (KE), 7.4 for retained fetal membranes (RFM), 4.5 for diarrhea (DI), 3.1 for displaced abomasum (DA), 2.9 for pneumonia (PN) and 1.9 for milk fever (MF). More than 30% of cows that had first cases of CM, DA, RFM, DI, and FD did not receive antibiotics. Of those treated, more than 50% of cows diagnosed with PN, ME and CM received ceftiofur as a treatment. The IR of mastitis and most other diseases was greater in older cows (parity ≥ 3) during the first 100 days of lactation and these cows were more likely to receive antibiotic treatments (as compared to younger cows diagnosed in later lactation). Cows of first and second parities in early lactation were more likely to remain in the herd after diagnosis of disease, as compared to older cows and cows in later stages of lactation. Most older cows diagnosed with CM in later lactation were culled before completion of the lactation. These results provide baseline data for disease incidence in dairy cows on modern U.S. dairy farms and reinforce the role of mastitis as an important cause of dairy cow morbidity.

The Impact of Selective Dry Cow Therapy Adopted in a Brazilian Farm on Bacterial Diversity and the Abundance of Quarter Milk.

We aimed to evaluate the impact of selective dry cow therapy (SDCT) (protocol 1: antimicrobial combined with internal teat sealant (ITS); vs. protocol 2: ITS alone) on bacterial diversity and the abundance of quarter milk. Eighty high production cows (parity ≤ 3 and an average milk yield of 36.5 kg/cow/day) from the largest Brazilian dairy herd available were randomly selected; milk quarter samples were collected for microbiological culture (MC) on the day of drying-off (n = 313) and on day 7 post-calving (n = 313). Based on the results of the MC before and after calving, 240 quarters out of 313 were considered healthy, 38 were cured, 29 showed new infections and 6 had persistent infections. Mammary quarters were randomly selected based on intramammary information status and SDCT protocols for bacterial diversity analyses. The bacterial diversity was similar when comparing both healthy and cured quarters submitted to both drying-off protocols. Despite healthy cows that were treated at dry-off using only teat sealant showing no alteration in the alpha and beta bacterial diversity, they did show a higher abundance of bacterial groups that may be beneficial to or commensals of the mammary gland, which implies that antibiotic therapy should be reserved for mammary quarters with a history of mastitis.

Birth conditions affect the longevity of Holstein offspring.

Studies of dairy cow longevity usually focus on the animal life after first calving, with few studies considering early life conditions and their effects on longevity. The objective was to evaluate the effect of birth conditions routinely collected by Dairy Herd Improvement agencies on offspring longevity measured as length of life and length of productive life. Lactanet provided 712,890 records on offspring born in 5,425 Quebec dairy herds between January 1999 and November 2015 for length of life, and 506,066 records on offspring born in 5,089 Quebec dairy herds between January 1999 and December 2013 for length of productive life. Offspring birth conditions used in this study were calving ease (unassisted, pull, surgery, or malpresentation), calf size (small, medium, or large), and twinning (yes or no). Observations were considered censored if the culling reason was "exported," "sold for dairy production," or "rented out" as well as if the animals were not yet culled at the time of data extraction. If offspring were not yet culled when the data were extracted, the last test-day date was considered the censoring date. Conditional inference survival trees were used in this study to analyze the effect of offspring birth conditions on offspring longevity. The hazard ratio of culling between the groups of offspring identified by the survival trees was estimated using a Cox proportional hazard model with herd-year-season as a frailty term. Five offspring groups were identified with different length of life based on their birth condition. Offspring with the highest length of life [median = 3.61 year; median absolute deviation (MAD) = 1.86] were those classified as large or medium birth size and were also the result of an unassisted calving. Small offspring as a result of a twin birth had the lowest length of life (median = 2.20 year; MAD = 1.69) and were 1.52 times more likely to be culled early in life. Six groups were identified with different length of productive life. Offspring that resulted from an unassisted or surgery calving and classified as large or medium when they were born were in the group with the highest length of productive life (median = 2.03 year; MAD = 1.63). Offspring resulting from a malpresentation or pull in a twin birth were in the group with the lowest length of productive life (median = 1.15 year; MAD = 1.11) and were 1.70 times more likely to be culled early in life. In conclusion, birth conditions of calving ease, calf size, and twinning greatly affected offspring longevity, and such information could be used for early selection of replacement candidates.

Erratum: Dallago et al. Keeping Dairy Cows for Longer: A Critical Literature Review on Dairy Cow Longevity in High Milk-Producing Countries. Animals 2021, 11, 808.

In the original article [...].

Keeping Dairy Cows for Longer: A Critical Literature Review on Dairy Cow Longevity in High Milk-Producing Countries.

The ability of dairy farmers to keep their cows for longer could positively enhance the economic performance of the farms, reduce the environmental footprint of the milk industry, and overall help in justifying a sustainable use of animals for food production. However, there is little published on the current status of cow longevity and we hypothesized that a reason may be a lack of standardization and an over narrow focus of the longevity measure itself. The objectives of this critical literature review were: (1) to review metrics used to measure dairy cow longevity; (2) to describe the status of longevity in high milk-producing countries. Current metrics are limited to either the length of time the animal remains in the herd or if it is alive at a given time. To overcome such a limitation, dairy cow longevity should be defined as an animal having an early age at first calving and a long productive life spent in profitable milk production. Combining age at first calving, length of productive life, and margin over all costs would provide a more comprehensive evaluation of longevity by covering both early life conditions and the length of time the animal remains in the herd once it starts to contribute to the farm revenues, as well as the overall animal health and quality of life. This review confirms that dairy cow longevity has decreased in most high milk-producing countries over time and its relationship with milk yield is not straight forward. Increasing cow longevity by reducing involuntary culling would cut health costs, increase cow lifetime profitability, improve animal welfare, and could contribute towards a more sustainable dairy industry while optimizing dairy farmers' efficiency in the overall use of resources available.

Herd-level associations between somatic cell counts and economic performance indicators in Brazilian dairy herds.

The aims of the present study were to provide a portrait of the techno-economic status of dairy herds in Minas Gerais, Brazil, particularly with respect to bulk-tank somatic cell count (BTSCC) data, and to examine the herd-level associations of BTSCC with various economic performance indicators (EPI). Data from 543 herds, 1,052 herd-year records in total, spread over 3 years (2015-2017), from the South and Southwest mesoregions of Minas Gerais State were provided by the Brazilian Support Agency to Micro and Small Companies Division Minas Gerais (SEBRAE). Herds had an average of 82 lactating cows per herd, milk yield of 17 L/cow per day, and availability of financial information via routine monthly economic surveys. The EPI data (revenue, gross margin, GM; net margin, NM; profit; break-even point; and operational profitability) of each herd was measured monthly by SEBRAE personnel, and herd-year averages of all variables were computed. Bulk-tank data (SCC, total bacterial count, content of crude protein and fat) taken by producers or dairy processors were recorded by SEBRAE personal; and corresponding herd-year averages were calculated and included in the SEBRAE database. There were 209 selected herds, which passed all edit checks, and which had data for all 3 years. The EPI (all expressed on a per-cow basis, $/cow per year) were analyzed, including the effects of region, year, log (ln) BTSCC, production level, and herd size, together with the random effect of herd nested within region. A high proportion of herds (94.6%) presented data records (herd-years) with an average BTSCC > 200 × 103 cells/mL: 37.8% of herd-year records had BTSCC between >200 and ≤400, 14.5% with BTSCC between >400 and ≤500, 25% with BTSCC between >500 and ≤750, and 17.3% with BTSCC >750. For each unit increase in ln BTSCC, revenue declined by $228.5/cow per year, GM by $155.6/cow per year, and profit by $138.6/cow per year. Herds with cows of lower production (<14 kg/d) presented lower GM ($286.8/cow per year) compared with herds containing cows producing ≥14 kg/d (≥14 and <19 kg/d = $446.5, and ≥19 kg/d = $601.9). The small-scale milk producers (<39 lactating cows) presented lower revenue ($1,914.9/cow per year) and GM ($274.5/cow per year) and consequently a negative profit (-$224.1/cow per year) compared with other herd size categories (≥39 lactating cows). The reduction in milk yield was 641 L/cow per lactation for each unit increase in ln BTSCC; this represented 9.4% of the milk yield per lactation, assuming an average milk production of 6,843.3 L/cow per lactation of cows from herds that had BTSCC ≤ 200 × 103 cells/mL. Consequently, we found a negative association of BTSCC with profit; profit declining from $227.0 to -53.1/cow per year when the BTSCC increased from 100 to 750 × 103 cell/mL. In short, the lower the BTSCC, the greater the revenue, GM and NM, profit, and operational profitability of the herds. The reduction of milk yield was the main factor associated with higher BTSCC.

Chronic subclinical mastitis reduces milk and components yield at the cow level.

We evaluated the effects of chronic subclinical mastitis (CSM) caused by different types of pathogens on milk yield and milk components at the cow level. A total of 388 Holstein cows had milk yield measured and were milk sampled three times at intervals of two weeks for determination of SCC and milk composition, and microbiological culture was performed. Cows were considered healthy if all three samples of SCC were ≤200 000 cells/ml and were culture-negative at the third milk sampling. Cows with one result of SCC > 200 000 cells/ml were considered to suffer non-chronic subclinical mastitis whereas cows with at least 2 out of 3 results of SCC > 200 000 cells/ml had CSM. These latter cows were further sorted according to culture results into chronic negative-culture or chronic positive-culture. This resulted in four udder health statuses: healthy, non-chronic, chronicNC or chronicPC. The milk and components yields were evaluated according to the udder health status and by pathogen using a linear mixed effects model. A total of 134 out of 388 cows (34.5%) were chronicPC, 57 cows (14.7%) were chronicNC, 78 cows (20.1%) were non-chronic and 119 cows (30.7%) were considered healthy, which resulted in a grand total of 1164 cow records included in the statistical model. The healthy cows produced more milk than each of the other groups (+2.1 to +5.7 kg/cow/day) and produced higher milk component yields than the chronicPC cows. The healthy cows produced more milk than cows with chronicPC caused by minor (+5.2 kg/cow/day) and major pathogens (+7.1 kg/cow/day) and losses varied from 5.8 to 11.8 kg/cow/day depending on the pathogen causing chronicPC mastitis. Chronic positive-culture cows had a reduction of at least 24.5% of milk yield and 22.4% of total solids yield.

Effect of transportation distances, seasons and crate microclimate on broiler chicken production losses.

The goal of this research was to evaluate the microclimate (temperature, relative humidity and ECI-enthalpy comfort index) of commercial loads of broiler chickens at different transport distances: Dist15 (15 km on average) and Dist90 (90 km on average) in the summer and winter seasons and their effects on the production parameters body weight difference (BWD), mortality (%) and bruising prevalence (%). Twelve broiler loads were monitored using dataloggers to record temperature and humidity, with a total of 24 target crates per load. The experiment followed a factorial design [2 seasons (rainy and dry) × 2 distances (Dist15 and Dist90)] with a randomized complete block arrangement, 3 sexes (all males, all females, or mixed shipments) and one shipment per combination. BWD had a heterogeneous distribution throughout the load, and this distribution was not significantly correlated with the mean ECI measured during transport at 12 positions along the load. In terms of comfort, summer is the most critical period for broiler transport. In the interaction between rainy season and Dist90, the highest ECI was scored in the lethal zone (where physiological mechanisms are not enough to control body temperature). Mortality during the rainy season was not significantly different between distances. However, during the dry season, mortality was twice as high as broilers that travelled for 15 km. The prevalence of bruising on carcasses was not affected by the interaction between season and distance. As we know, broiler chicken performance, during transport, can be also related to road conditions, being hard to evaluate the real impact of seasons and distances on animal welfare. Load microclimate can compromise broiler chicken welfare during transport and it does not necessary reflect significant losses pre and post-slaughter.

Correlations between the Composition of the Bovine Microbiota and Vitamin B12 Abundance.

Vitamin B12 is synthesized by prokaryotes in the rumens of dairy cows-and this has implications in human nutrition since humans rely on consumption of dairy products for vitamin B12 acquisition. However, the concentration of vitamin B12 in milk is highly variable, and there is interest in determining what causes vitamin B12 variability. We collected 92 temporally linked rumen, fecal, blood, and milk sample sets from Holstein cows at various stages of lactation fitted with rumen cannula and attempted to define which bacterial genera correlated well with vitamin B12 abundance. The level of vitamin B12 present in each sample was measured, and the bacterial population of each rumen, fecal, and milk sample (n = 263) was analyzed by 16S rRNA-targeted amplicon sequencing of the V4 region. The bacterial populations present in the rumen, small intestine, and milk were highly dissimilar. Combined diet and lactation status had significant effects on the composition of the microbiota in the rumen and in the feces. A high ruminal concentration of vitamin B12 was correlated with the increased abundance of Prevotella, while a low ruminal concentration of vitamin B12 was correlated with increased abundance of Bacteroidetes, Ruminiclostridium, and Butyrivibrio The ultimate concentration of vitamin B12 is controlled by the complex interaction of several factors, including the composition of the microbiota. Bacterial consumption of vitamin B12 in the rumen may be more important in determining overall levels than bacterial production.IMPORTANCE In this paper, we examined the microbiome of the bovine rumen, feces, and milk and attempted to understand how the bacterial communities at each site affected the production and movement of vitamin B12 around the animal's body. It was determined that the composition of the bovine rumen microbiome correlates well with vitamin B12 concentration, indicating that the rumen microbiota may be a good target for manipulation to improve production of this important vitamin.

Milk ß-hydroxybutyrate concentration measured by Fourier-transform infrared and flow-injection analyses from samples taken at different times relative to milking.

Analysis of milk BHB concentration by Fourier-transform infrared (FTIR) spectrometry more frequently than regular milk testing could help dairy producers in decision making, particularly if it would be possible to use small hand-stripped samples (hereinafter simply called samples) taken between dairy herd improvement (DHI) test-samples analysed using DHI algorithms. The aim of this Research Communication was to evaluate milk BHB concentration and the prevalence of elevated milk BHB concentration analysed by FTIR spectrometry compared with flow-injection analysis (SKALAR) from samples taken at different times relative to the milking. A total of 293 early-lactation cows in 44 commercial dairy herds were involved in the study. Herds were visited once during the morning milking when a routine DHI test-sample was obtained using in-line milk samplers. Additional milk samples were taken by hand stripping as follows: (1) Just before connecting the milking machine; (2) immediately after removing the milking machine; (3) 3 h after milking and (4) 6 h after milking. Milk samples were analysed for BHB concentration by FTIR and SKALAR, the latter being the reference method. Milk BHB concentration from samples taken before milking was different between FTIR and SKALAR whereas no difference was noted for other sampling times, although milk BHB concentration rose as time after milking increased. Except for DHI test-samples for which prevalence was not different between analysis methods, prevalence of elevated milk BHB concentration (≥0.15 mmol/l) was greater for FTIR analysis. However, no difference in prevalence was observed between SKALAR and FTIR when using a threshold of ≥0.20 mmol/l. In summary, hand-stripped milk samples taken any time after removing the milking machine until 6 h after the milking can be recommended for FTIR analysis of elevated milk BHB concentration prevalence provided a threshold of 0.20 mmol/l is used.

Milk losses associated with somatic cell counts by parity and stage of lactation.

The reduction of milk production caused by subclinical mastitis in dairy cows was evaluated through the regression of test-day milk yield on log-transformed somatic cell counts (LnSCC). Official test-day records (n = 1,688,054) of Holstein cows (n = 87,695) were obtained from 719 herds from January 2010 to December 2015. Editing was performed to ensure both reliability and consistency for the statistical analysis, and the final data set comprised 232,937 test-day records from 31,692 Holstein cows in 243 herds. A segmented regression was fitted to estimate the cutoff point in the LnSCC scale where milk yield started to be affected by mastitis. The statistical model used to explain daily milk yield included the effect of herd as a random effect and days in milk and LnSCC as fixed effects regressions, and analyses were performed by parity and stage of lactation. The cutoff point where milk yield starts to be affected by changes in LnSCC was estimated to be around 2.52 (the average of all estimates of approximately 12,400 cells/mL) for Holsteins cows from Brazilian herds. For first-lactation cows, milk losses per unit increase of LnSCC had estimates around 0.68 kg/d in the beginning of the lactation [5 to 19 d in milk (DIM)], 0.55 kg/d in mid-lactation (110 to 124 DIM), and 0.97 kg/d at the end of the lactation (289 to 304 DIM). For second-lactation cows, milk losses per unit increase of LnSCC had estimates around 1.47 kg/d in the beginning of the lactation (5 to 19 DIM), 1.09 kg/d in mid-lactation (110 to 124 DIM), and 2.45 kg/d at the end of the lactation (289 to 304 DIM). For third-lactation cows, milk losses per unit increase of LnSCC had estimates around 2.22 kg/d in the beginning of the lactation (5 to 19 DIM), 1.13 kg/d in mid-lactation (140 to 154 DIM), and 2.65 kg/d at the end of the lactation (289 to 304 DIM). Daily milk losses caused by increased LnSCC were dependent on parity and stage of lactation, and these factors should be considered when estimating losses associated with subclinical mastitis.

Effects of herbicides and the chytrid fungus Batrachochytrium dendrobatidis on the health of post-metamorphic northern leopard frogs (Lithobates pipiens).

Effects of exposure to contaminants such as pesticides along with exposure to pathogens have been listed as two major contributors to the global crisis of declining amphibian populations. These two factors have also been linked in explanations of the causes of these population declines. We conducted a combined exposure experiment to test the hypothesis that exposure to two agricultural herbicides would increase the susceptibility of post-metamorphic northern leopard frogs (Lithobates pipiens) to the amphibian fungal pathogen Batrachochytrium dendrobatidis (Bd). We assessed the independent and interactive effects of these exposures on the health and survival of the frogs. Wild-caught frogs underwent a 21-day exposure to a nominal concentration of either 2.1 µg/L atrazine (Aatrex(®) Liquid 480) or 100 µg a.e./L glyphosate (Roundup(®) Original), followed by Bd, and then were observed until 94 days post-initial exposure to the herbicides. Actual levels of atrazine were between 4.28 ± 0.04 µg/L and 1.70 ± 0.26 µg/L while glyphosate degraded from 100 µg a.e./L to approximately 7 µg a.e./L within 6 days of initial exposure to the herbicides. Compared to controls, the glyphosate formulation reduced the snout-vent length of frogs during the pesticide exposure (at Day 21), and the atrazine formulation reduced gain in mass up to Day 94. No treatment affected survival, splenosomatic or hepatosomatic indices, the densities and sizes of hepatic and splenic melanomacrophage aggregates, the density and size of hepatic granulomas, proportions of circulating leucocytes, the ratio of neutrophils to lymphocytes, or the ratio of leucocytes to erythrocytes. Histological assessment of samples collected at Day 94 revealed no evidence of Bd infection in any Bd-exposed frogs, while real-time PCR detected only one case of light infection in a single atrazine- and Bd-exposed frog. Frogs exposed to Bd shed their skin significantly more frequently than Bd-unexposed frogs, which may have helped them resist or clear infection, and could explain why no interaction between the herbicides and Bd was detected. The results suggest that these frogs were resistant to Bd infection and that pre-exposure to the herbicides did not alter this resistance. The effects seen on the growth following herbicide exposure is a concern, as reduced growth can lower the reproductive success and survival of the amphibians.

Identification and quantitation of reaction intermediates and residuals in lipase-catalyzed transesterified oils by HPLC.

A high-performance liquid chromatography (HPLC) unit equipped with size exclusion column and a refractive index detector was used for simultaneous monitoring, identification, and quantitation of the reaction components from lipase-catalyzed transesterification of three oils. The procedure simultaneously separated and detected the unreacted triacylglycerols (TAG), diacyl-, and monoacyl-glycerol (DAG and MAG) co-products, residual alcohol as well as free fatty acid (FFA) based on retention times. The chromatograms showed well separated and resolved peaks. The elution of the components from the transesterification reaction in increasing order was: TAG < DAG < FFA < MAG. Generally, higher alcohol ratios decreased the conversion of TAG in all the oils studied with between 14% and 94% of TAG remaining at all the treatment combinations. Higher amount of salmon skin oil (SSO) TAG was generally converted to DAG than Rothsay composite (RC) and olive oil (OO) TAG. Relatively higher amount of OO DAG was converted to MAG than SSO and RC with only 5-14% DAG remaining in OO. RC and OO generally accumulated less MAG, and this was reflected as lower MAG levels in RC (<6%) and OO (<14%) compared with SSO (<27%). For the various treatment combinations and the three oils used in this study, the least amount of FFA was recorded in transesterified OO with a maximum of approximately 4%. This HPLC method can be used as a simple and fast technique to analyze the reaction components and products of transesterification reactions without the need for additional derivatization steps.