Progress 04/30/21 to 08/31/21
Outputs
Target Audience:The target audiences are livestock producers, Animal Science and Veterinary students, Veterinarians, Diagnosticians, Toxicologists, and Regulatory officials. Changes/Problems:Additional work is needed to determine whether it is safe to feed 200 ppm bromide in hay.? What opportunities for training and professional development has the project provided?This project provided numerous opportunities for training and for professional development. Conducted during COVID-19 outbreak when opportunities for research were extremely limited, this project provided employment for undergraduate students in the Animal Science Program who participated to earn internship hours or for experience. About 15 undergraduate students contributed to the project feeding cows, monitoring cows during birth, assessing and recording health of cows and calves. They also helped bottle feed the weak calves. About 8 Veterinary students in their first year volunteered to assist on the projects. With regard to professional development, this project has been very impactful. One of the veterinary students presented an abstract on the project at the national annual AAVLD conference in Denver, CO. The PI has presented results of this project at the Society of Toxicology and AAVLD meetings. In addition, scientists from Iowa State University who optimized an assay to quantify bromide in tissues, presented posters at the AAVLD meeting. How have the results been disseminated to communities of interest?Several abstracts have been published and presentations given at national scientific toxicology and veterinary diagnostic toxicology meetings. Current effort is to publish the work in peer-reviewed journals. What do you plan to do during the next reporting period to accomplish the goals?In the next reporting period we plan to report on outcome of analysis of samples from calves. We posit that since calves are more sensitive, results of tissue bromide analysis in calves in conjunction with hormone analysis and histopathology of the endocrine glands, especially the thyroid glad will guide answer the question whether feeding bromide at 200 ppm to pregnant cows in the 3rd trimester is safe.
Impacts
What was accomplished under these goals?
Major goals of the project Anecdotal field reports had indicated that pregnant cows and their newborn calves are very sensitive to inorganic bromide intoxication. There were no studies reported in the literature on effects of inorganic bromide in pregnant cows and on new born calves exposed in utero. USDA funded this novel project to fulfil two objectives: to determine the level of bromide within the total mixed ration (TMR) that could safely be fed to pregnant cows without negatively impacting the health of the pregnant cows and of fetuses; and to determine bromide residue levels in blood, liver, kidney, urine, milk and muscle of cows and calves afterfeeding them bromide contaminated hay. This objective will address the safety of cattle products to consumers. The study showed that inorganic bromide fed in concentrations exceeding 800 ppm in total mixed ration jeopardized the health on the new born calves exposed in utero. The study confirmed field observations that calves exposed in utero are more sensitive to inorganic bromide intoxication. Calf deaths were reported in the 800 and 3200 ppm groups. Calves in these high dose groups were born weak, had week suckling reflexes and had to be bottle fed. These calves could not stand and find the udder. The consequence of this outcome is that without intervention affected calves would not get colostrum. Therefore, they would be susceptible to infection later in life and likely not survive. We also successfully measured bromide tissue residues in cows. We established normal bromide tissue concentrations. These results were inconclusive as to whether 200 ppm inorganic bromide in total mixed ration was safe for to feed to cattle as additional tissue analysis, including those of calves was needed.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2021
Citation:
Schrunk D, Ivanytska O, Jensen A, Clapham MC, Tell L, Rumbeiha WK. Bromide analysis in Bovine plasma and tissue by ICP-MS. Proceedings of the AAVLD Annual Meeting, Denver, CO
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2021
Citation:
Mastorakis AN, McNabb B, Lima F, Urbano T, Tell L, Rumbeiha KW. In utero inorganic bromide exposure causes locomotor and behavioral abnormalities in newborn calves. Proceedings of the AAVLD annula meeting, Denver CO.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2022
Citation:
Rumbeiha WK, Tell L, Flaws J, Watson KD, Kim D. Characterizing a large animal model of bromide-induced endocrine disruption in neonates. The Toxicologist. A publication of the proceedings of the SOT conference.
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Progress 04/30/21 to 08/31/21
Outputs
Target Audience:The target audiences for this project were: 1) USDA which needed to confirm that inorganic bromide is toxic to pregnant cows. USDA also needed to know what is the highest non toxic dose in hay if it was toxic; 2) cattle producers who had made cursory field observations and had linked consumption of bromide contaminated hay to difficulties in calving, assisted parturition, and weak calves. They also needed confirmation that problems observed in their farms were indeed caused by inorganic bromide intoxication. Beyond these two key audiences, results of this project are of interest to veterinarians and veterinary students. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Three undergraduate student in Animal Science participated as interns which was required for their degree completion. One veterinary student presented this work at AAVLD. The PI has attended and presented this work at two international conferences *AAVLD and the Annual Society of Toxicology meeting). Eight first year veterinary students volunteered on the project to gain practical experience in veterinary toxicology. How have the results been disseminated to communities of interest?This work has been presented at 2 internatiional conferences. A manuscript is in preparation for publication in peer reviewed literature. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
Results: Key findings from the study are now briefly summarized. All 20 cows, except Tag #1 in the control group which aborted a dead fetus, delivered live calves. Except for 2/4 (50%) of cows in the 3200-ppm group, all cows which went to full term calved unassisted. For the 2 cows in the 3200-ppm group that were assisted, calves were pulled by a veterinarian following prolonged labor (> 2-3 hrs). Moderate effort was used to pull calves. The rest of the cows presented with normal labor (calved within 1 hour of labor). Two cows gave birth to twins: Tag 20 from the 200-ppm group and Tag 18 from the 3200-ppm group. The cause of abortion in the 1 cow from the 0-ppm control group was determined to be uterine fungal infection. In this experiment calves were the most impacted by bromide exposure. Two calves from bromide-fed cows died, Tag 7A from the 800-ppm group and Tag 10A from the 3200-ppm group. They died within 10 days of birth from colitis. No other calves died. The most outstanding observation was that 6/6 (100%) of the calves in the 3200-ppm group and 1/4 calves (25%) in the 800-ppm group were born very lethargic, depressed, and had difficulty to stand. Whereas calves in the control group stood and walked within 22 to 88 mins after birth, one calf in the 50 and one calf in the 200 ppm groups stood and walked after 88 mins. Remarkably, 75% of the calves in the 800 ppm and 100% of the calves in the 3200-ppm group stood after 88 mins. One calf in the 3200-ppm group stood after 822 mins (13.7 hrs).In parallel to weakness and inability to stand after birth, the most severely affected calves had very weak suckling reflexes. One calf from the 800-ppm group and the rest of the calves in the 3200-ppm group were bottle fed colostrum and milk replacer because they could not suckle. It is important to note that calves need to suckle colostrum from their dams within 4 hrs of birth in order to get optimum immunoprotection. This was impaired in some of the 800-ppm dose group and all of calves in the 3200-ppm group. Three factors accounted for inability to suckle: 1) inability to stand; 2) weak suckle; and behavioral impairment (disorientation) with inability to find the udder. Calves were disorientated and ataxic for 2-4 days after birth, unable to access the udder necessitating bottle feeding or intubation for as long as 4 days after birth. There was a dose-dependent trend in suppression of calf growth rate starting with 200 ppm group and with the calves in the 3200-ppm group most affected. However, this data was not statistically significant, likely because of the small sample size of calves in this study. Nonetheless, growth suppression starts with the 200-ppm group. In cows, we observed prolonged labor, hind limb ataxia and behavioral impairment. Prolonged labor was noted in 2 of 4 cows in the 3200-ppm group. Naturally, cows have a natural instinct to groom their calves at birth to keep them clean. They also have a natural instinct to be protective of their calves. Cows in the 800 and 3200 ppm groups lost aggressiveness against caretakers, which instinct was strong in cows in the rest of the groups. Cows in the high dose groups also did not care to groom their calves as shown in Tag 18 from the 3200-ppm group. Plasma Chemistry A limited panel (pH, bicarbonate, TCO2, calcium, glucose, creatinine, blood urea nitrogen, sodium, potassium, and chloride) was performed in calves at birth. Abnormalities were noted in only two parameters i.e., chloride and glucose. All calves in the control group (0 ppm bromide) had chloride and glucose concentrations within normal ranges (97-109 mEq/l and 56-74 mg/100 mL respectively). Also, all calves from the 50 ppm groups had both chloride and glucose in normal ranges. In the 200-ppm group however, chloride was marginally elevated in one calf (21a) at 112 mEq/L. Strikingly, chloride was severely elevated in 50% of calves in the 800-ppm group and in 100% of calves in the 3200-ppm group. This is called pseudo hyperchloremia because most analyzers misread bromide as chloride. Pseudo hyperchloremia has been reported in cases of bromide intoxication but this is the first report of this condition in new borne calves. As for glucose, hypoglycemia was noted in some calves. Plasma glucose concentration was below normal range (56-74 mg/100 mL) only in the 3200-ppm dose group. In this group, blood glucose concentrations were in the range of 38-42 mg/100 mL a few hours after birth. Hormone Analysis Results We measured progesterone and pregnancy specific protein B in cows only. The plasma concentration of progesterone was not different between control and any bromide treatments (50 PPM, 200 PPM, 800 PPM, and 3200 PPM) at enrolment (P = 0.99), 28 days after treatment (P = 0.77), 56 days after treatment (P = 0.99), and 84 days after treatment (P = 0.91). Similarly, the plasma concentration of pregnancy specific protein B was not different between control and any bromide treatments (50 PPM, 200 PPM, 800 PPM, and 3200 PPM) at enrolment (P = 0.27), 28 days after treatment (P = 0.60), 56 days after treatment (P = 0.57), and 84 days after treatment (P = 0.56). These results suggest that inorganic bromide exposure did not affect hormonal and pregnancy status in cows. Plasma Bromide Results Normal blood plasma bromide concentration was < 10 ppm in control (0 ppm) and in cows of all other groups at the start of the study (baseline). However, plasma bromide concentration was still above normal range in all bromide fed cows at the time the study was terminated on day 30 post-calving. Although plasma bromide results were interesting, the most dramatic results were found in calves (Fig. 8). In all bromide-fed groups plasma bromide concentration was consistently higher in calves than in cows at the time the experiment was terminated. For example, on average, plasma bromide concentration in the 3200 group cows was about 100 ppm; yet in calves it was > 1000 ppm (normal is < 10 ppm). Another interesting observation was that the calf: cow plasma bromide ration was at per (1:1) at parturition/birth but the ratio was much higher at study termination on day 30. Specifically, the calf/cow ratio of plasma bromide in various groups was 3.67, 7.50, 11.37, and 13.86 respectively for the 50, 200, 800, and 3200 ppm groups. This suggests either: a) calves have difficulty excreting bromide; b) calves were still exposed to bromide through dam milk during the 30-day period; or c) both. A limited data set from this study indicated that bromide is passed in milk in a dose-dependent manner. In tissues normal bromide concentration was < 10 ppm. No bromide residues were found in calf tissues (> 10 ppm), including muscle, 30 days post-calving in the 50-ppm group. Bromide concentration in muscle was > 10 ppm in the 200 and 800 ppm groups. In cowsbromide residue concentration in muscle was within normal range (< 10 ppm) in all groups except the 3200-ppm group. Notably, the trend of the ratio of tissue bromide in calves to that in cows is similar to that in blood plasma i.e., higher in calf tissues at euthanasia than in cow. euthanasia. Organ Weights There were no statistically significant differences in tissue organ weights of cows exposed to bromide compared to control although thyroid glands of cows in the 3200-ppm group were generally larger in weight compared to the other groups. In calves, there was a statistically significant increase in calf adrenal weight in the 3200-ppm group compared to the 50-ppm group. Although not statistically different, the weights of calf thyroid glands were smaller in calves from bromide administered groups than controls. There were no other changes in organ weights.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2022
Citation:
The Toxicologist. Proceedings of the Society of Toxicology meeting 2022.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2022
Citation:
Proceedings of the Annual AAVDL meeting in minneapolis, MN
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