Objectives were to evaluate the effects of an oral supplement containing soluble Ca, and live yeast in LPS-challenged dairy cows. The trial consisted of 2 experimental periods (P). During P1 (3 d), cows (n = 12) were fed ad libitum and baseline data was collected. At the beginning of P2 (which lasted 96 h), all cows were i.v. challenged with 0.375 mu g/kg BW LPS. Cows were assigned randomly to 1 of 2 treatments: 1) control (CON; no bolus; n = 6) or 2) an oral bolus containing Ca and live yeast (CLY; YMCP Vitall (R) 44.718 g of elemental Ca; TechMix, LLC., Stewart, MN; n = 6), administered -0.5 and 6.5 h relative to LPS infusion. Following LPS administration, circulating Ca decreased in both treatments but supplemental CLY ameliorated the hypocalcemia (48 h area under the curve: -10.8 vs. -1.9 mmol/L x h; P < .01). Lipopolysaccharide decreased dry matter intake (DMI; 60%) similarly for both treatments on d 1, but overall (d 1-4) DMI tended to be reduced less (14 vs. 30%; P = .06) in CLY supplemented vs CON cows. Lipopolysaccharide reduced milk yield (70%; P < .01) from 12 to 24 h, but throughout P2, milk yield from CLY supplemented cows was increased (38%; P = .03) relative to CON cows. Overall during P2, circulating LPS-binding protein and serum amyloid A increased post LPS (3- and 4-fold, respectively, P < .01), but were unaffected by treatment (P >= .68). In conclusion, providing an oral supplement containing Ca and live yeast prior to and following LPS administration markedly ameliorated LPS-induced hypocalcemia and improved DMI and milk yield.

Objectives were to determine the effects of a product containing electrolytes, osmolytes, and energetic compounds (EOEC) on body temperature indices in heat-stressed (HS) Holstein cows. Lactating cows were assigned to 1 of 2 treatments: 1) a control diet (n = 10) or 2) a control diet supplemented with 113 g/d of EOEC (n = 10; Bovine BlueLite® Pellets; TechMix LLC, Stewart, MN). The trial consisted of 2 experimental periods (P). During P1 (4 d), cows were fed their respective treatments and housed in thermoneutral conditions. During P2 (4 d), HS was artificially induced using an electric heat blanket (EHB). Overall, HS markedly increased vaginal temperature (Tv), rectal temperature (Tr), skin temperature (Ts), and respiration rate (RR) (P < .01). There were no dietary treatment differences in Tv, Tr, or RR; however, during P2 EOEC-supplemented cows had increased Ts (0.8 °C; P = .04). Compared to P1, HS decreased DMI and milk yield (45 and 27%, respectively, P < .01) similarly amongst treatments. Relative to P1, circulating insulin decreased (41%; P = .04) in CON cows, whereas it remained unaffected in EOEC-supplemented cows, resulting in a 2-fold increase in EOEC compared with CON-fed cows (P < .01) during P2. Relative to P1, HS increased circulating non-esterified fatty acids (NEFA; 63%; P < .01). During P2, there tended to be a treatment by day interaction on circulating NEFA, as concentrations decreased from d 2 to 4 of P2 in EOEC-fed cows but continued to increase in CON cows. In summary, feeding EOEC altered some key aspects of energetic metabolism and increased Ts.

The objective of this study was to evaluate the effects of supplementing a Saccharomyces cerevisiae fermentation product (SCFP) on body temperature indices, metabolism, acute phase protein response, and production variables during heat stress (HS). Twenty multiparous lactating Holstein cows (body weight = 675 ± 12 kg; days in milk = 144 ± 5; and parity = 2.3 ± 0.1) were used in an experiment conducted in 2 replicates (10 cows/replicate). Cows were randomly assigned to 1 of 2 dietary treatments: control diet (CON; n = 10) or the CON diet supplemented with 19 g/d of SCFP (n = 10; NutriTek, Diamond V, Cedar Rapids, IA). Cows were fed their respective diets for 21 d before initiation of the study. The experiment consisted of 2 periods: thermoneutral (period 1; P1) and heat stress (period 2; P2). During P1 (4 d), cows were fed ad libitum and housed in thermoneutral conditions for collecting baseline data. During P2 (7 d), HS was artificially induced using an electric heat blanket (EHB; Thermotex Therapy Systems Ltd., Calgary, AB, Canada). Cows were fitted with the EHB for the entirety of P2. Rectal temperature, respiration rate, and skin temperature were obtained twice daily (0600 and 1800 h) during both periods. Overall, HS increased rectal temperature, skin temperature, and respiration rate (1.4°C, 4.8°C, and 54 breaths/min, respectively) relative to P1, but no dietary treatment differences were detected. Compared with P1, HS decreased dry matter intake and milk yield (36 and 26%, respectively), and the reductions were similar between dietary treatments. Relative to P1, HS increased milk fat content and milk urea nitrogen (17 and 30%, respectively) and decreased milk protein and lactose contents (7 and 1.4%, respectively). Overall, HS increased (52%) plasma cortisol concentrations of CON, but circulating cortisol did not change in SCFP-fed cows. Heat stress increased circulating lipopolysaccharide binding protein and serum amyloid A (SAA; 2- and 4-fold, respectively), and SCFP supplementation tended to decrease peak SAA (∼33%) relative to CON cows. Overall, although HS did not influence circulating white blood cells and neutrophils, SCFP increased circulating white blood cells and neutrophils by 9 and 26%, respectively, over CON in P2. In conclusion, HS initiated an acute phase protein response and feeding SCFP blunted the cortisol and SAA concentrations and altered some key leukocyte dynamics during HS.

On page 9637, the final sentence of the Results should read (correction shown in bold): “There was a treatment × period interaction on circulating platelets, as they remained unchanged in CON cows but decreased in SCFP-fed cows during P2 (31%; P = 0.05; Table 5).” Effects of a Saccharomyces cerevisiae fermentation product on heat-stressed dairy cowsJournal of Dairy ScienceVol. 103Issue 10PreviewThe objective of this study was to evaluate the effects of supplementing a Saccharomyces cerevisiae fermentation product (SCFP) on body temperature indices, metabolism, acute phase protein response, and production variables during heat stress (HS). Twenty multiparous lactating Holstein cows (body weight = 675 ± 12 kg; days in milk = 144 ± 5; and parity = 2.3 ± 0.1) were used in an experiment conducted in 2 replicates (10 cows/replicate). Cows were randomly assigned to 1 of 2 dietary treatments: control diet (CON; n = 10) or the CON diet supplemented with 19 g/d of SCFP (n = 10; NutriTek, Diamond V, Cedar Rapids, IA). Full-Text PDF Open Access

The efficacy of an electric heat blanket (EHB) has previously been confirmed as an alternative method to evaluate heat stress (HS). However, a pair-feeding design has not been used with the EHB model. Therefore, study objectives were to determine the contribution of the nutritional plane to altered metabolism and productivity during EHB-induced HS. Multiparous Holstein cows (n = 18; 140 ± 10 d in milk) were subjected to 2 experimental periods (P); during P1 (4 d), cows were in thermoneutral conditions with ad libitum feed intake. During P2 (4 d), cows were assigned to 1 of 2 treatments: (1) thermoneutral conditions and pair-fed (PF; n = 8) or (2) EHB-induced HS with ad libitum feed intake (n = 10). Overall, the EHB increased rectal temperature, vaginal temperature, skin temperature, and respiration rate (1.4°C, 1.3°C, 0.8°C, and 42 breaths/min, respectively) relative to PF cows. The EHB reduced dry matter intake (DMI; 47%) and, by design, PF cows had a similar pattern and extent of decreased DMI. Milk yield decreased in EHB and PF cows by 27.3% (12.1 kg) and 13.4% (5.4 kg), respectively, indicating that reduced DMI accounted for only ∼50% of decreased milk synthesis. Milk fat content tended to increase (19%) in the EHB group, whereas in the PF cows it remained similar relative to P1. During P2, milk protein and lactose contents tended to decrease or decreased (1.3 and 2.2%, respectively) in both EHB and PF groups. Milk urea nitrogen remained unchanged in PF controls but increased (34.2%) in EHB cows relative to P1. The EHB decreased blood partial pressure of CO2, total CO2, HCO3, and base excess levels (17, 16, 17, and 81%, respectively) compared with those in PF cows. During P2, the EHB and PF cows had similar decreases (4%) in plasma glucose content, but no differences in circulating insulin were detected. However, a group by day interaction was detected for plasma nonesterified fatty acids; levels progressively increased in PF controls but remained unaltered in the EHB cows. Blood urea nitrogen increased in the EHB cows (61%) compared with the PF controls. In summary, utilizing the EHB model indicated that reduced nutrient intake explains only about 50% of the decrease in milk yield during HS, and the postabsorptive changes in nutrient partitioning are similar to those obtained in climate-controlled chamber studies. Consequently, the EHB is a reasonable and economically feasible model to study environmental physiology of dairy cows.

Rumination involves a complex series of muscle contractions that bring a bolus of ingesta to the mouth for further mastication before it is swallowed again. Healthy cows ruminate 8 to 9 h/d. Hypocalcemia is known to disrupt nerve and muscle function. Our hypothesis was that hypocalcemia in periparturient cows would reduce rumination activity. Twenty-six Holstein cows entering their third lactation or greater were fed a control diet [dietary cation-anion difference (DCAD) = +196 mEq/kg of dry matter (DM)] or a low DCAD diet supplemented with anions (DCAD = -9 mEq/kg of DM) prepartum. Periparturient plasma Ca concentration and rumination rate were determined. Four of 12 control cows developed clinical milk fever, necessitating intravenous Ca therapy. Rumination rate decreased in all cows around the time of calving. Rumination rate on the first and second day of lactation was highly correlated with the cow's plasma Ca concentration on the first day of lactation. In one of our statistical models, a normocalcemic cow was defined as a cow whose plasma Ca concentration remained above 2.00 mM. Cows were retrospectively classified as normocalcemic, subclinically hypocalcemic, or clinically hypocalcemic (milk fever). Only 4 cows were considered normocalcemic, and all had been fed the low DCAD diet. Normocalcemic cows spent more time ruminating on the first day after calving than subclinically hypocalcemic cows or cows with milk fever. Cows with milk fever had a lower rumination rate than normocalcemic cows through d 3 of lactation. Rumination activity in cows with milk fever was almost nondetectable in the hours before and after intravenous Ca treatment for an extended period, despite the return of muscle function that allowed the cows to stand and eructate following treatment. Other statistical models using different definitions of normocalcemia gave qualitatively similar results. Diet had a great effect on plasma Ca concentration and rumination rate. Even when cows with clinical milk fever were removed from the control cow data set, cows on the low DCAD diet had significantly greater plasma Ca concentrations in the first 36 h after calving and a higher rumination rate on d 1 of lactation (248 ± 26 min) than control cows (158 ± 32 min).

Precisely studying the biological consequences of heat stress (HS) in agriculturally relevant animals typically requires expensive climate-controlled facilities, infrastructure inaccessible to most researchers. Thus, study objectives were to explore the efficacy of an electric heat blanket (EHB) as an alternative method for evaluating HS and to determine whether EHB-induced hyperthermia affects production parameters similar to natural HS. Lactating Holstein cows (n = 8; 133 ± 3 d in milk; 709 ± 31 kg; 2.6 ± 0.3 parity) were housed in individual box stalls and allowed to acclimate for 3 d. After acclimation, the trial consisted of 2 experimental periods (P). During P1 (3 d), cows were housed in thermoneutral conditions for collecting baseline data. During P2 (7 d), cows were fitted with an EHB. During the entire experiment cows were fed ad libitum, and dry matter intake (DMI) was recorded daily. Cows were milked twice daily (0600 and 1800 h), and milk samples were collected on d 2 and 3 of P1 and d 3 and 7 of P2. Rectal temperature, respiration rate, heart rate, and skin temperature were obtained twice daily (0600 and 1800 h) during both P1 and P2. Overall, there was an increase in rectal temperature and respiration rate at 0600 h (1.0°C and 25 breaths/min, respectively) and 1800 h (1.2°C and 29 breaths/min, respectively) during P2. The EHB decreased DMI and milk yield (25 and 21%, respectively) by the end of P2. During P2, milk protein tended to decrease (4.4%) compared with P1. In contrast, milk urea nitrogen increased (33%) during P2 relative to P1. No other differences were observed in milk composition. In summary, our results indicate that employing an EHB affects physiological and production parameters similarly to natural HS (i.e., increased rectal temperature and respiration rate, decreased DMI and milk yield); thus, the EHB is an effective and inexpensive research tool for evaluating the biological consequences of HS in lactating dairy cows.

Background Coagulase negative Staphylococcus (CNS) species are currently the most prevalent intra-mammary pathogens causing subclinical mastitis and occasional clinical mastitis or persistent infection in lactating dairy cattle. More than 10 CNS species have been identified, but they are generally managed as one group on most dairies in the United States. However, improved management decisions and treatment outcomes may be achieved with better understanding of the prevalent species, pathogenicity and strain diversity within and across dairies. Methodology A total of 604 CNS isolates were cultured from milk samples collected during a dry-cow treatment clinical trial conducted on 6 dairy herds in 4 states in the US. All the study cows were randomized to receive 1 of the 3 different intra-mammary antimicrobial infusions (Quatermaster, Spectramast DC or ToMorrow Dry Cow) at dry-off. Milk samples were collected at dry-off, calving (0–6 days in milk, DIM), post-calving (7–13 DIM) and at mastitis events within the first 100 DIM. The CNS isolates were identified to species level by partial sequencing of the rpoβ gene, and genetic relatedness within species was investigated by phylogenetic analysis of the pulse-field gel electrophoresis profiles of the isolates. Results The major CNS species identified were S. chromogenes (48.3%), S. haemolyticus (17.9%), S. simulans and S. epidermidis (each at 6.5%). Other CNS species identified at lower frequencies included S. hominis, S. auricularis, S. sciuri, S. spp KS-SP, S. capitis, S. cohnii, S. warneri, S. pasteuri, S. xylosus, S. hyicus, S. equorum, S. microti, S. rostri, S. gallinarum, S. saprophyticus and S. succinus. Phylogenetic analyses of the major species types demonstrated an association between genetic relatedness and epidemiological distributions of S. chromogenes, S. simulans, S. haemolyticus and S. auricularis. Additionally, identical strains of S. chromogenes and S. simulans were isolated from the same udder quarter of several cows at consecutive sample stages. The rest of the minor species had no deducible genetic-epidemiological link. Discussion The observed association between genetic and epidemiological distributions indicated animal-adapted nature of four CNS species, suggesting possible host-adapted and environmental transmission of these species. Multi-stage isolation of the same udder quarter strain was evidence for chronic intra-mammary infection. Conclusion The different CNS species and strains circulating on US dairy herds were genetically diverse. Four species identified were likely udder-adapted pathogens, 2 of which caused persistent infection. Our findings are important in guiding the design of effective mastitis control strategies.

Abstract Heat stress (HS) negatively impacts production parameters, including milk yield and composition, growth and reproduction. Precisely studying HS typically requires expensive climate-controlled facilities, resources often inaccessible to most scientists. Therefore, we evaluated the efficacy of using an electric heat blanket (EHB) as an alternative and cost-effective method to study HS and determine whether EHB-induced hyperthermia affects physiological and production parameters similar to natural HS. This was the first proof of concept study examining this model. Results indicated increased body temperature indices (i.e., rectal temperature and respiration rate) and reduced DMI and milk yield, as well as altered milk composition similar to natural and climate-controlled HS studies. In experiment 2, we examined the EHB model in combination with a pair-feeding (PF) design. By employing the PF design, we clearly illustrated that a lowered nutritional plane explains only approximately 50% of the decreased milk yield in HS cows, with the remaining portion due to changes in post-absorptive changes in nutritional physiology. In addition, the EHB blunted adipose tissue mobilization and increased plasma biomarkers of muscle catabolism. Results from this study validated that the EHB is a valid model to study HS; therefore, experiment 3 was designed to assess nutritional HS mitigation strategies using the EHB model. Dietary electrolyte, osmolyte, and energetic compounds (EOEC) were evaluated as re-hydration therapy on body temperature indices in HS lactating Holstein cows. Skin temperature was increased in EOEC-supplemented cows relative to controls which suggests that EOEC increased heat dissipation. In addition, EOEC-supplemented cows had increased glucose and insulin levels compared with controls which is due to the energetic compounds (i.e., dextrose) present in the EOEC supplement. Collectively, employing EHB model provides an excellent new platform for discovery research and evaluating pragmatic HS mitigation strategies.

Administrating lipopolysaccharide (LPS) decreases circulating calcium (Ca) and markedly reduces both feed intake and milk yield in lactating cows. Calcium is involved in immune system activation and live yeast can increase feed intake. Whether supplemental live yeast and Ca benefits immune-challenged cows remains unclear. Therefore, study objectives were to evaluate if providing an oral supplement containing soluble Ca, live yeast and other micronutrients would ameliorate LPS-induced hypocalcemia and production parameters in lactating dairy cows. Providing an oral supplement containing Ca and live yeast prior to and following LPS administration markedly ameliorated the LPS-induced hypocalcemia and improved DMI and milk yield. Overall, utilizing an oral supplement may be a valuable management strategy to improve animal welfare and productivity during and following immunoactivation. Additionally, infusing i.v. LPS appears to be an effective technique to model hypocalcemia and to evaluate dietary strategies aimed at increasing circulating calcium in lactating dairy cows.

The goal of dipping the umbilical cord after birth in calves is to promote healing of the umbilical stump, prevent infection, and encourage the umbilical tissue to detach from the body. Treatment applied to the umbilical area is an important management step for preventing morbidity and mortality in calves. The objective of this study was to compare the effect of 4 umbilical dips on the healing rate, incidence of infection, and age at umbilical cord detachment using newborn Holstein heifer calves (n = 73). Calves were alternately assigned by birth order to 4 treatment groups: 7% iodine, a dry dip formulated using an antibacterial peptide (nisin) mixed with talc (3.105 g of nisin per 100 g of talcum powder on a weight per weight basis), liquid nisin (64 µg/mL), and 4% chlorhexidine mixed with alcohol in a 50:50 solution. Umbilical cords were dipped 30 min after birth. Before initial dipping, umbilical cord diameter (as an indicator of the rate of cord drying and healing rate) was determined using a digital caliper. The caliper measurements were repeated at 24 ± 1, 48 ± 1, and 72 ± 1 h (±standard deviation) of age and were continued daily until the umbilical cord healed and detached from the animal's body. Diagnosed umbilical infections were documented by veterinary staff based on a combination of clinical symptoms (redness, swelling, purulent discharge, painful response (flinch or kicking) to palpation of the umbilical stump) in addition to a lack of appetite and fever. Data were analyzed using MIXED model procedures with fixed effect of umbilical treatment. No treatment differences were noted between dips on the umbilical cord drying rate or days for umbilical cord to detach. Treatment effects were observed on incidence of umbilical infection (incidence of infection for calves across all treatments was 9.0%).

Mastitis research has shown 40-50% of intramammaryinfections (IMI) are contracted during the dry or non-lactating period with greatest percentages occurring during first and last two weeks of dry period. The ability to develop and apply external persistent barrier teat dip products that can persist for these 1 week periods could decrease IMI, thus improving animal health and performance, and product quality and safety.Objective of this study was to evaluate an experimentalsealant product vs. commercial persistent barrier dry cow teatsealant dip with particular interest and comparisons of dip persistency in providing teat end protection, and overall teat end and skin health. Two external teat sealants were applied to 24 animals for assessment of adherence to teatskin/teat end over a period of fifteen days. Overall, substantially better coverage wasobserved for teats treated with the experimental product compared to control product over thefirst week after application. By the third day, the experimental productwas 22 times more likelyto have teats protected than the control product. By day four, this likelihood increased to 56times. Product was observed on teats with the experimental productuntil day 10, while noproduct was visible by day 5 on teats treated with the control product. The studyshowed thatthe experimental productstayed on teats much longer than the control product, with around50% teat ends protected five days after initial application.

Heat stress (HS) is an annual environmental issue which negatively affects a variety of production parameters including milk yield and composition, growth, and reproduction. A study was conducted toexplore the efficacy of an electric heat blanket (EHB) as an alternative method to study HSand to determine whether EHB-induced hyperthermia affects production parameters similar to natural HS.During this study, the animals were fitted with an ear tag based behavior and temperature monitor (Cow Manager, Agis Automatisering, Netherlands) toevaluate behavior and ear temperature changes through the heat stress study and understand the potential use of behavior monitors in the field for early heat stress identification and mitigation. Also 24 hr continuous time lapse video was captured on all animals during their time in the trial barn. Data from this trial is being analyzed. This paper presents behavior and temperature graphs, data, and comments from one animal over her trial period. When animals were moved to the trial barn, ear temperatures increased to 80-950F and remained there for the 2 week barn period indicating increased blood flow to the ear for heat dissipation. Ear temperatures were highest during the blanketed period. There were some daily and diurnal changes associated with external temperatures (barn had curtains). Mostlyanimals rested with heads toward outside cooler walls, evidencing animal behavior to mitigate heat load. Ear temperatures dropped immediately and returned to normal when returned to colder herd free stall barn. Compared to baseline daily normal activity % (eating, ruminating, non-active, active, and highly active), changes were seen when animal were moved to the barn or in the barn (increased activity, perceived increased eating but related to more head movement due to environment adjustments rather that truly eating). During week 1 in the barn (warmer barn but no blankets), behavioral changes captured included decreased eating times (15-18% vs. 25-27% normally) but uniquely increased rumination times (which were associated with increased respiration and panting activity rather than rumination). Once the blanket was applied for 1 week, further behavioral changes were seen with daily eating times decreased to 8-10% (also changes when animals ate -mostly only when moved to milk) but rumination times increased (65-75%) but was associated with increases in respiration and panting movement rather than rumination. When blankets were removed and animals returned to normal freestall, ear temperatures immediately adjusted and dropped to normal but behavior changes to normal took 1-6 days. Overall, the electric blanket heat stress model was successful and behavior and ear temperature monitors were very successful in quantifying changes associated with heat stress (earlywith slightly warm temperatures through higher temperatures associated under the blanket). Accelerometers were excellent in measuring movement but newer algorithms need to separate random head movements from true eating, and mouth movements with no other movements associated with panting from rumination to improve diagnostic performance. Overall, the monitors look like an excellent technology to observe changes in early moderate heat stress and provide a tool for implementing early heat stress management and mitigation strategies.

An eight week trial was conducted to evaluate the teat conditioning efficacy of oneexperimental iodine post milking teat dip formula compared to a commercial iodine product.Teat skin and teat end roughnesswere scored for each teat 2X/week. A total of 48 cows were scored during thestudy period, but only 36 cows had full records for analyses. Results showed noconcerning teat skin condition irritation events. There were no differences betweenproducts in teat end condition. However it was observed that teat end conditionworsened over time for both products, most likely as a result ofchanges intemperature during winter.

A 4 week trial was conducted at the ISU dairy to evaluate the teat conditioning efficacy of two experimental organic acid teat dip formulas. Teat skin and teat end thickness and roughness were scored on each teat 2X/week. Results showed that teat skin was maintained at the optimum level (score 1) on all teats of all cows for both experimental products over the full trial period. Teat end condition remained stable (and excellent) during the test period and no significant differences were observed between treatments, scoring dates or the interaction treatment*score date. It is concluded that both experimental products have similar teat condition efficacy properties.

Heat stress (HS) is an annual environmental issue which negatively affects a variety of production parameters including milk yield and composition, growth, and reproduction.However, precisely studying HS typically requires expensive climate controlled facilities; infrastructure inaccessible to most researchers.Thus, study objectiveswere to explore the efficacy of an electric heat blanket (EHB) as an alternative method to study HSand to determine whether EHB-induced hyperthermia affects production parameters similar to natural HS.Utilizing the EHB increased body temperature indices (rectal temperatures and respiration rate)and reduced dry matter intakeand milk yield. Our results indicate that employing the EHB affects production parameters similarly to natural HS and thus the EHB is an effectiveandinexpensive research tool to evaluate the biological consequences of HS in lactating dairy cows.

Mastitis research has shown that 40-50% of intramammaryinfections (IMI) are contracted during the dry or non-lactating period with the greatest percentages of these occurring during the first and last two weeks of the dry period. The ability to develop and apply external persistent barrier teat dip products (like a liquid bandage) that can persist for these 1 week periods could decrease IMI, thus improving animal health and performance, and product quality and safety.The objective of this study was to evaluate and compare 3persistent barrier dry cow teatsealant dips with particular interest and comparisons of dip persistency in providing teat end protection, and overall teat end and skin health. Cows dipped with alldipshad significantly greater persistency and protection compared to previous 4 trials (last 2 years).All dips were easy to apply and showed excellent teat health. Films were very consistent and very flexible over time (limited ripping, shredding or flaking).Overall, Dip C showed greatest persistency in the first 5-7days post dipping which is a very critical time period.