Monitoring selected behaviors of calves by use of an ear-attached accelerometer for detecting early indicators of diarrhea

One of the most important diseases in calves worldwide is neonatal calf diarrhea (NCD), which impairs calf welfare and leads to economic losses. The aim of this study was to test whether the activity patterns of calves can be used as early indicators to identify animals at risk for suffering from NCD, compared with physical examination. We monitored 310 healthy female Holstein-Friesian calves on a commercial dairy farm immediately after birth, equipped them with an ear tag–based accelerometer (Smartbow, Smartbow GmbH), and conducted daily physical examinations during the first 28 d of life. The Smartbow system captured acceleration data indicative of standing and lying periods and activity levels (active and inactive), shown as minutes per hour. We categorized calves as diarrheic if they showed fecal scores of ≥3 on a 4-point scale on at least 2 consecutive days. Incidence of diarrhea was 50.7% (n = 148). A mixed logistic regression model showed that lying [odds ratio (OR) = 1.19], inactive (OR = 1.14), and active (OR = 0.92) times, 1 d before clinical identification of diarrhea (d ?1), were associated with the odds of diarrhea occurring on the subsequent day. Receiver operating characteristics curve showed that lying time at d ?1 was a fair predictor for diarrhea on the subsequent day (area under curve = 0.69). Average lying time on d ?1 was 64.8 min longer in diarrheic calves compared with their controls. Median lying and inactive times decreased, and active time increased with age over the study period. The 24-h pattern of behavior indices based on the output of the Smartbow system followed periods of resting and active times, and showed that between 2200 h and 0600 h, calves spent the greatest percentage of time lying and inactive. These results showed that the accelerometer system has the potential to detect early indicators associated with NCD. In future studies, additional data for the development and testing of calf- and event-specific algorithms (e.g., for detecting milk intake, playing behavior) should be collected, which might further improve the early detection of diarrhea in calves.     

Effect of preweaning disease on the reproductive performance and first-lactation milk production of heifers in a large dairy herd

Raising replacement heifers represents a substantial cost in dairy farms, with reproductive efficiency being one of the main factors driving the total rearing cost. Diseases during the preweaning period, such as diarrhea or bovine respiratory disease (BRD), are reported at high incidence risks worldwide. However, the long-term effects of disease before weaning on productive and reproductive performance remain controversial. This retrospective cohort study explored the extent to which diseases such as diarrhea or BRD during the preweaning period affected average daily gain (ADG), herd removal, reproductive indices, and 305-d mature equivalent milk production (305ME) of replacement heifers. The health, growth, and production records of 2,272 female calves from a large dairy herd were used; 487 and 926 of the calves had BRD and diarrhea, respectively, before weaning. The reproductive variables age at first service, age at successful service, age at calving, and pregnancies per artificial insemination were calculated. Associations of disease status with reproduction indices, removal from herd, ADG, and 305ME were evaluated by survival analysis and mixed models. Heifers with a history of BRD before weaning were less likely to be inseminated or achieve first calving than heifers without BRD. However, BRD status did not change the age at first insemination of calving among those being inseminated or reaching first calving, respectively. There were also no differences in ADG, 305ME, and pregnancies per artificial insemination between heifers with and without a history of BRD. A history of diarrhea preweaning was not associated with changes in the probability of being inseminated, having a confirmed pregnancy, or reaching first calving. However, heifers affected by diarrhea required more inseminations to become pregnant. Diarrhea status was also associated with a 325-kg reduction in 305ME in the first lactation and a 50 g/d reduction in ADG. Collectively, our results demonstrate some long-term effects of diseases before weaning on the reproductive efficiency of heifers and first-lactation 305ME production, but with limited effect on the time to achieve critical reproductive performance indicators.     

Health benefits of orally administered anti-IL-10 antibody in milk-fed dairy calves

The primary objective of this randomized controlled trial was to determine whether anti-IL-10 egg yolk antibodies fed upon arrival to a calf ranch would lower the prevalence of Cryptosporidium parvum shedding in naturally challenged preweaned dairy calves. The secondary objectives included measuring the effect of anti-IL-10 antibodies on calf health, performance, and shedding of less common diarrheal pathogens. A total of 133 calves, enrolled at 24 to 72 h of age, received a daily dose of 0.96 g of egg yolk powder with anti-IL-10 antibodies (MAB, n = 71) or without anti-IL-10 antibodies (MEP, n = 62) split between 2 feedings for the first 11 d on feed at a calf ranch. Daily health evaluations were completed for 15 d after arrival and on d 56. Digital weights were collected at enrollment and d 56, and hipometer weights were collected at enrollment and d 7 and 56. Packed cell volume and serum total protein concentration were measured at enrollment and on d 7 and 14. Fecal pH was measured at enrollment and on d 5 and 14, and fecal pathogen (C. parvum, coronavirus, rotavirus, and Salmonella spp.) shedding was assessed at d 5 and 14. Continuous outcomes were compared between groups using a Student's t-test or Wilcoxon rank sum test. Fecal pathogen shedding at d 14, respiratory disease at d 56, and antibiotic usage were compared using relative risk (RR) and chi-squared test. Fecal pH (median and interquartile range) on d 14 was 6.65 (6.39–6.99) and 6.52 (5.97–6.81) for MAB and MEP, respectively. On d 56, the risk of respiratory disease was lower for MAB compared with MEP (RR = 0.40; confidence interval = 0.16–0.99). The risk for antibiotic treatment was lower for MAB- compared with MEP-treated calves (RR = 0.38; confidence interval = 0.17–0.88). The risk of shedding rotavirus was higher in MAB (RR = 1.38; confidence interval = 1.10–1.81) calves. After multivariable analyses, hipometer weights (least squares means ± standard error) were 1.7 ± 0.8 kg greater on d 56 in MAB compared with MEP; however, ADG was 0.04 ± 0.02 kg/d lower in MAB calves. Total health score, diarrhea days, average respiratory score, packed cell volume, and serum total protein were not affected by feeding anti-IL-10 egg antibodies. In summary, feeding anti-IL-10 antibodies was associated with increased fecal pH, reduced risk of respiratory disease later in the preweaning period, and decreased antibiotic usage despite higher rotavirus infection. These findings might be associated with improved mucosal immunity, enhanced host defenses, or reduced susceptibility and warrant further investigation.     

Short communication: Effect of pomegranate-residue supplement on Cryptosporidium parvum oocyst shedding in neonatal calves

Cryptosporidium parvum is considered one of the most common enteropathogens, responsible for the high incidence of diarrhea and deleterious implications on immunity and health in neonatal calves. The pomegranate is well known for its health-promoting properties. Two experiments were designed to test the antiparasitical and antidiarrheal effects of concentrated pomegranate extract (CPE) supplement in milk in neonatal Holstein calves. Forty-one calves were randomly divided into control (n = 20) and treatment (n = 21) groups. For the first experiment, the treatment group was supplemented with 3.75% CPE in the daily milk ration, between 3 and 14 d of age, whereas the control group received only milk. Fecal samples were collected between d 5 and 13 to quantify Cryptosporidium oocysts, and the duration and intensity of diarrhea were evaluated. Reduced fecal oocyst count and diarrhea intensity and duration were revealed in the 3.75% CPE calves. No difference was noted in average daily gain between groups. In a second experiment, which was designed to test the effect of a lower CPE concentration (0.6% of daily milk allocation), no effects on fecal oocyst count and average daily gain were observed. However, compared with control, the lower CPE group was characterized by a shorter duration of diarrhea and higher weight gain among males at 14 d of age. These results suggest that the CPE supplement-to-calf milk ratio may potentially alleviate intestinal morbidity caused by Cryptosporidium.     

Nutraceutical intervention with colostrum replacer: Can we reduce disease hazard,ameliorate disease severity,and improve performance in preweaned dairy calves?

The objective of this randomized clinical trial was to determine if an intervention with colostrum replacer (CR; Premolac Plus, Zinpro; 125 g/d fed for 3 d) or a placebo milk replacer (Cows Match, Land O'Lakes Inc.; 125 g/d for 3 d) following a triggered alarm could ameliorate disease bouts in dairy calves. The alarm was set to detect negative deviations of milk intake (20% reduction) or drinking speed (30% reduction) in relation to a calf's 12-d rolling average feeding behavior. Calves were enrolled on this study (n = 42 CR, n = 42 placebo) when they triggered an alarm from d 14 to 50 on the feeder before weaning. Once calves were enrolled, calves received a bottle of either 125 g of CR or milk replacer mixed with 1 L of water for 3 d. Calves were enrolled on the automated milk feeder at age 4.0 ± 2.0 d (mean ± SD), were offered 10 L/d of the same milk replacer fed as a placebo, and had ad libitum access to calf starter measured by automated feeders. Weaning began at d 50, with 50% reduction of milk replacer across 14 d, and an additional 20% reduction for another 7 d before weaning at d 70. Calves were health scored from birth to 2 wk postweaning (88 ± 2.0 d of age) daily for bovine respiratory disease (BRD) and diarrhea, and were weighed and scored using lung ultrasonography twice weekly. A BRD score ≥5 and an area of consolidated lung ≥3.0 cm² was considered BRD positive; diarrhea was defined as a watery fecal consistency that sifted through the bedding. The effect of CR on BRD and diarrhea likelihood were calculated with logistic models; the 7 d before and after an alarm were fixed effects, with milk intake as a covariate. A Cox proportional hazards model evaluated the effect of CR on the hazard of being positive for BRD and diarrhea following enrollment, with sex as a fixed effect in the diarrhea model. The effect of CR on average daily gain the week following intervention was evaluated with linear modeling with repeated measures, and birth date was a random effect. After intervention, placebo calves had 1.64 (95% CI: 1.11–2.43) times greater odds of having a BRD bout compared with CR calves for the 7 d following intervention. Moreover, placebo calves had 1.50 (95% CI: 1.11–2.08) times greater odds of having lobar lung consolidation in the 7 d after intervention than CR calves. No difference was found in the likelihood of diarrhea in the 7 d after intervention (odds ratio: 0.91; 95% CI: 0.71–1.16). However, placebo calves had a 2.38 (95% CI: 1.30–4.33) times greater hazard of incurring BRD in the 14 d after intervention. Average daily gain was not associated with an intervention with CR for the 7 d after intervention (placebo: 0.73 ± 0.07 kg/d; CR: 0.70 ± 0.08 kg/d; LSM ± SEM). These results suggest CR may reduce BRD likelihood but did not influence growth in calves offered high allowances of milk. Future research should examine which properties of CR ameliorate BRD in calves.     

Effects of ad libitum milk intake on dairy calves

Conventional milk feeding for calves (by bucket twice daily at a total of 10% of body weight) was compared with feeding milk for ad libitum consumption from a nipple. Calves were weaned gradually between d 37 and 42 by diluting the milk with water, and body weight and feed consumption were followed until d 63. Calves fed ad libitum drank 89% more milk than calves fed conventionally during the preweaning period, but the ad libitum-fed calves ate only 16% as much calf starter and 17% as much hay. Consumption of starter and hay increased rapidly after weaning, and treatment differences disappeared. Probably as a result of the much higher intake of milk, the ad libitum-fed calves gained 63% more weight than the conventionally fed calves before weaning, resulting in a 10.5-kg weight advantage on d 35. During and immediately after weaning, the rate of weight gain slowed for both treatment groups, but recovered by approximaely d 49. There were no treatment differences in weight gains over the weaning or postweaning periods, and at the end of the experiment on d 63, the calves fed ad libitum maintained an advantage in mean (+/- SEM) body weight (89.07 +/- 2.47 kg vs 81.07 +/- 2.47 kg for the conventionally fed calves). Incidence of diarrhea was low and did not differ between treatment groups. We conclude that ad libitum nipple feeding of milk to dairy calves can allow for increased milk intake and weight gain with no detrimental effects on intake of solid food after weaning.     

The effect of neomycin inclusion in milk replacer on the health,growth, and performance of male Holstein calves during preweaning

The prophylactic use of oral antimicrobials, such as neomycin, in milk replacer (MR) or whole milk is a common practice in calf-rearing that is thought to aid in preventing disease. Heavy reliance on antimicrobials is of concern not only because of the development of antimicrobial resistance, but also because of the potentially negative effects on health. The objective of this study was to investigate the effects of neomycin on calf health and growth performance. One hundred and sixty calves (approximately 3–10 d of age), distributed across 2 experimental periods, were stratified by body weight (BW) and serum total protein, and assigned to 1 of 3 treatments: control (CON; nonmedicated MR, n = 60), short-term antibiotic (ST; neomycin mixed in MR from d 1–14, n = 50), or long-term antibiotic (LT; neomycin in ?MR from d 1–28, n = 50). Arrival BW (47.69 ± 0.87 kg) and serum total protein (5.67 ± 0.09 g/dL) were not different between treatment groups. Neomycin in ST and LT was dosed in MR at a rate of 20 mg/kg of BW and was adjusted weekly according to BW. Calf BW was measured weekly for 49 d, and health indicators (fecal score, attitude score, respiratory score, and rectal temperature), MR intake, starter intake, and the use of additional electrolytes and antimicrobials were recorded daily. Calves in the CON group experienced a higher proportion of days with diarrhea (20.32 ± 0.02%) compared with ST (14.70 ± 0.02%) or LT (13.80 ± 0.02%) calves, as well as longer bouts of diarrhea (7.45 ± 0.38 d, 5.69 ± 0.46 d, and 5.62 ± 0.45 d for CON, ST, and LT calves, respectively). Calves in the CON group also experienced higher fecal scores (score of 0.64 ± 0.04) than ST (score of 0.53 ± 0.04) or LT (score of 0.49 ± 0.04) calves, especially at d 7. However, no differences were observed in other health-related measures. The time to reach first diarrhea and first respiratory illness was not different between treatments, nor was the time to recover from respiratory illness. The time to intervention with additional electrolytes or antimicrobials was not different between treatment groups. Furthermore, growth performance, feed intake, and feed conversion ratio were not different. No differences were found when comparing ST and LT, except in the defined daily dose of total antimicrobials received. Calves in the LT group received a higher overall dose than ST calves, and both ST and LT calves received a higher dose than CON calves, which received no prophylactic antimicrobials. Given that there were no differences in performance variables and no additional health benefits aside from reduced fecal scores in calves fed neomycin, current practices involving the use of antimicrobials on dairy and veal operations need to be considered more prudently.     

猪流行性腹泻病毒冻干qLAMP检测体系的建立

为建立猪流行性腹泻病毒（Porcine Epidemic Diarrhea Virus,PEDV）实时荧光定量LAMP（Quantitative LAMP,qLAMP）与冻干技术结合的快速检测方法。该实验根据PEDV的高度保守序列设计了引物并进行筛选,建立PEDV的qLAMP检测方法。筛选棉子糖、甘露醇和牛血清蛋白作为冻干辅料并优化,将最佳的配方与等温扩增试剂（除甜菜碱）混合后进行冷冻干燥处理。优化主干燥升温方式和二次干燥时间,制作出一条适合等温扩增试剂的冻干曲线,并以此制备PEDV qLAMP冻干检测试剂盒。结果显示：通过对冻干成品性状、溶解性及剩余水分含量的测定,确定了主干燥后期采用一步升温的方式进行升温,解析干燥时间为3.5 h。PEDV qLAMP冻干检测体系的最低检测限为每微升157 copies,与工艺优化前相比,其检测速率得到提升。对猪瘟病毒、猪伪狂犬病毒等5种病原无扩增反应,重复性实验变异系数均小于5%,常温条件下保存30 d,在4 ℃条件下至少保存12个月。该研究通过使用冻干技术建立了LAMP检测方法,具有现场操作简便快捷、检测效果好、储存时间长,适用于基层现场快速检测。     

北京市水产品污染与感染病例中副溶血性弧菌血清型和毒力基因型的比较研究

目的对北京市夏季市售水产品污染与感染病例中副溶血性弧菌血清型和毒力基因型进行比较研究,为评估食品安全风险监测的目的与意义提供思路,为北京市水产品副溶血性弧菌污染与临床感染病例的关联性研究提供技术支持。方法对采集的水产品样品和哨点医院腹泻患者粪便样本进行副溶血性弧菌的分离鉴定,采用血清玻片凝集法对分离出的副溶血性弧菌进行血清分型,PCR方法检测菌株的tlh、tdh、trh基因。结果 2014年7~9月共采集水产品样品164份,检出副溶血性弧菌80份,总污染率为48.78%;其中淡水产品污染率为38.78%(19/49),平均菌量浓度为66.63 MPN/g;海水产品污染率为53.04%(61/115),平均菌量浓度为38.14 MPN/g。80株副溶血性弧菌分属于9个血清群,其中O2群28株,占35.00%,O1群11株,占13.75%,O5群10株,占12.50%。80株菌tlh基因均为阳性,只有1株菌携带tdh毒力基因,所有菌株trh毒力基因均为阴性。哨点医院腹泻病人粪便样本中分离鉴定副溶血性弧菌21株,血清型O3∶K6占61.90%(13/21),O4:K8占28.57%(6/21);毒力基因型tdh(+)/trh(-)占95.24%(20/21),tdh(-)/trh(-)占4.76%(1/21)。结论来源于食品样品的副溶血性弧菌绝大部分不具备致病性,而导致消费者腹泻的副溶血性弧菌绝大部分携带致病性毒力基因,表明目前的食品安全风险监测结果不能作为评估副溶血弧菌导致的食源性疾病暴发和散发的依据。     

广州某生猪养殖场大肠杆菌流行性及抗生素的耐药性

本研究采集广州某规模化养殖场腹泻仔猪、健康仔猪及环境样本共147份,据国标GB 4789.6-2016分离鉴定大肠杆菌,用K-B法分析分离株对12种抗生素的敏感性,并用PCR检测其对应的耐药基因。结果共检出大肠杆菌115株;分离菌株依次对复方新诺明,左氧氟沙星、加替沙星、环丙沙星呈高水平耐药,耐药率30%;对庆大霉素、头孢他啶、氨曲南也呈现一定水平耐药(20%~30%);对多粘菌素B、替加环素、亚胺培南耐药水平较低(15%),对美罗培南敏感;46.09%菌株呈现多重耐药(≥3种抗生素),且腹泻仔猪源菌株多重耐药率显著高于健康仔猪源及环境源菌(p0.05)。分离株耐药基因携带严重,其中ant(3')-Ⅰa、aac(3')-Ⅰb、sul1、aac(3)-Ⅱb,blaTEM、blaCTX、tet M基因检出率50%,因此,这可能是本研究菌株的主要耐药机制。研究结果表明该养殖场大肠杆菌污染严重,耐药水平高,有关部门需加强养殖中腹泻仔猪防治用药的监管与指导。