Emerging technologies for identifying superior dairy cows in New Zealand

The performance of animals is determined by the interaction of their genes with environmental circumstances. Accordingly, animals exhibiting superior performance are not necessarily the animals with the best genes nor are they the best choice of parents. Statistical analyses of production records for repeated traits, e.g. lactation yields and reproductive performance, show that part of the variation in performance among animals in the same herd and year is due to genetic differences, and the remainder is due to so-called residual or environmental factors that are not passed on to offspring. These within-herd environmental factors can be partitioned into a component that affects performance throughout an animal's lifetime, and a part that is unique to each observation. The process of animal evaluation from pedigree and performance records partitions the superiority of each cow into these three components. Reliable assessment of the genetic merit of bulls has required progeny testing, and for cows has required observation of their own individual performance. Selection on the genetic or breeding value component has systematically improved animal performance over recent decades, but has been limited by the age at which assessments of genetic merit are available. Emerging molecular technologies can read DNA sequences or measure RNA expression and have allowed the identification of a number of chromosome regions, and a few specific genes in those regions, that influence economic performance. This information allows better characterisation of the relationships between animals and more accurate predictions of genetic merit in bulls without progeny information and in cows that have yet to produce their own performance record. At some stage, enough genes responsible for variation in performance will be identified to allow faster genetic progress through selection of animals at young ages and therefore more rapid turnover of the generations. Mechanisms that modify gene expression have been identified and these may ultimately allow animals to be selected at an early age for lifetime productivity, accounting for processes that modify gene expression and lead to differences in performance that are not reflected by DNA sequence information. This review describes the status of these emerging technologies and their likely role in the improvement of dairy cattle.     

Breeding dairy cows for the future in New Zealand

A brief history of the breeding of dairy cattle in New Zealand is provided. Dairy farming in New Zealand is unique compared with the majority of dairy systems in the developed world. New Zealand has a dependence on grass-based diets and a strict requirement for a 365-day calving interval. Four main areas are discussed: future traits to evaluate, advances in genetic evaluation technologies, impacts of crossbreeding, and future progeny testing schemes. These areas are not independent, e.g. the trend of increasing numbers of crossbred cattle in the national herd will have major impacts on the design of breeding schemes. It is foreseeable that in the future there will be improvements in the national breeding goal to better reflect on-farm profitability, and in the definition of traits and methods of data capture within the national breeding goal. Methods of selection and genetic evaluation that are currently feasible for a small population will become feasible for large populations as computing power improves. Genetic improvement of cows in New Zealand will continue to be a critical component of the increased economic efficiency achieved on dairy farms in this country.     

Risk factors for and reproductive outcomes of phantom cows on New Zealand dairy farms

AIMS: To determine some of the risk factors for cows not observed in oestrus within 35–42 days of an unsuccessful artificial insemination (AI; phantom cows), and the reproductive outcomes and effect of treatment of phantom cows.

MATERIALS AND METHODS: Over 2 years, in dairy herds from the Waikato (n=10) and Canterbury (n=4) regions of New Zealand, pregnancy diagnosis was carried out 35–42 days after AI on cows that had been inseminated in the first 3 weeks after the start of mating (PSM) but had not been seen returning to oestrus. Risk factors for phantom cows were analysed using a generalised linear mixed effect model.

In Year 1, all phantom cows were left untreated. In Year 2, phantom cows were categorised as having a corpus luteum (CL) (CL+ n=120), or having ovarian follicles ≥10 (n=101) or <10 (n=40)?mm in diameter. Cows with a CL were treated with cloprostenol or untreated and placed with bulls. Cows with no CL received intravaginal progesterone (P4) for 7 days, with injection of gonadotrophin-releasing hormone (GnRH) on Days 0 and 9, and cloprostenol on Day 7 followed by AI. Pregnancy diagnosis of all cows took place 100–120 days after PSM and interval to conception and final pregnancy rate determined.

RESULTS: Overall, of cows inseminated in the first 3 weeks after PSM that did not return to oestrus, 610/6,734 (9.1%) were phantom cows. From the final multivariable analysis, treatment for anoestrus, BCS ≤4.0 at mating, being 2 or >6 years of age, and pure-bred, and decreasing interval between calving and mating, until 98 days post calving, were associated with increased odds of being a phantom cow. Compared to all other groups of cows, phantom cows had a longer interval to conception (p<0.001) and a lower final pregnancy rate (p<0.001).

Treatment of CL+ cows or cows with follicles ≥10?mm did not affect reproductive outcomes (p>0.3). For cows with follicles <10?mm treatment decreased the final percentage not pregnant (3/27; 11%; p=0.01) and interval to conception (21 days; p=0.02) compared with controls (7/13; 54% and 37 days, respectively).

CONCLUSIONS AND CLINICAL RELEVANCE: Risk factors for phantom cows were identified that could be manipulated to reduce the number of phantom cows in a herd, in particular increasing BCS. Treatment of the majority of phantom cows did not improve reproductive performance.     

Genetic and crossbreeding parameters for incidence of recorded clinical lameness in New Zealand dairy cattle

AIM: To estimate genetic and crossbreeding parameters for the incidence of recorded clinical lameness in New Zealand dairy cattle.

METHODS: Herd records from 76,357 cows, collected during the 2005/06 to 2008/09 milking seasons from 155 herds in the Livestock Improvement Corporation young sire progeny test scheme, were used to estimate genetic parameters and breed effects for incidence of recorded clinical lameness in HolsteinFriesian, Jersey and crossbred dairy cattle. Recorded clinical lameness was coded “1” for cows that presented at least one event of clinical lameness at any day during the season and “0” for unaffected cows. Genetic parameters were estimated using an animal model across breeds considering all and then only first lactation records. Heritability and repeatability of recorded clinical lameness were calculated from the variance component estimates both with and without logit transformation.

RESULTS: The mean incidence of recorded clinical lameness per herd was 6.3 (min 2, max 34)%. The incidence of recorded clinical lameness in Holstein Friesian cows (mean 6.8, SE 0.24%) was higher than the incidence of recorded clinical lameness in crossbred (mean 6.1, SE 0.19%) and Jersey cows (mean 6.0, SE 0.28%) (p=0.0002). There was no difference in incidence between crossbred and Jersey cows (p=0.96).

Estimates of the heritability of recorded clinical lameness as an untransformed trait were 0.053 (SE 0.014) for first lactation records and 0.016 (SE 0.003) for all lactation records. As a transformed (logit) trait heritabilities were 0.067 (SE 0.024) and 0.044 (SE 0.016) for first and all lactation records, respectively. The repeatability estimates of recorded clinical lameness were 0.071 (SE 0.005) and 0.107 (SE 0.011) for untransformed and logit transformed lactation records, respectively. Sire estimated breeding values for recorded clinical lameness showed the lowest values in Jersey sires, and ranged between -5 and 8%.

CONCLUSIONS: Despite the low heritability of recorded clinical lameness, this study provided evidence that there is significant exploitable animal genetic variation. Selection of specific sires across and within breeds could be an option for increasing genetic resistance to lameness in New Zealand dairy cattle.     

Resynchronising returns to service in anoestrous dairy cows in the South Island of New Zealand

AIMS: To investigate the effect of targeted resynchronisation of cows treated for non-observed oestrus before the planned start of mating (PSM), that were not detected in oestrus or pregnant 23 days after treatment (phantom cows), on the proportion pregnant at 42 days after PSM and the end of mating.

METHODS: Farm staff from eight herds in two regions of the South Island of New Zealand identified 1,819 cows not showing oestrus by 10 days before PSM. These cows were treated with intravaginal progesterone for 7 days, and I/M gonadorelin 10 days and 1 day before PSM. Three days before PSM they were injected with cloprostenol and equine chorionic gonadotrophin, with fixed time artificial insemination (FTAI) at PSM. By 23 days after PSM, 1,218 cows had not returned to oestrus. Of these, 161 cows confirmed not pregnant by transrectal ultrasonography were randomly assigned to no treatment (control group; n=74) or were resynchronised 25 days after PSM using the same treatment programme as above, with FTAI 35 days after PSM (n=87). All cows that returned to oestrus were artificially inseminated until 42 days after PSM, when natural mating was used. All cows were examined using transrectal ultrasonography 80 to 90 days after PSM to confirm conception dates.

RESULTS: Of the 1,819 anoestrous cows treated before PSM, 526 (29 (95% CI=23.1–34.0)%) had not been observed in oestrus by 23 days after PSM and had not conceived, so were diagnosed as phantoms cows. For resynchronised cows, 42/87 (48 (95% CI=37.8–58.8)%) were pregnant by 42 days after PSM compared to 21/74 (28 (95% CI=18.1–38.7)%) control cows (p=0.009). At the end of mating 58/87 (67 (95% CI=56.6–76.7)%) cows in the resynchronised group were pregnant and 46/74 (62 (95% CI=50.9–73.2)%) in the control group (p=0.554). The hazard of conception from 21 to 42 days after PSM was 1.9 (95% CI=1.07–3.12) times greater for resynchronised than control cows (p=0.026).

CONCLUSION: In cows not observed in oestrus and treated before PSM, resynchronisation increased the proportion pregnant by 42 days after PSM.

CLINICAL RELEVANCE: The benefit of resynchronisation depends on the number of anoestrous cows before PSM and the number of phantom cows after PSM. However at the herd-level it is likely that providing advice to reduce the known risk factors for cows not being observed in oestrus before the PSM may well be more cost effective than identifying and treating a sub-population of phantom cows.     

Signals for identifying cows at risk of subacute ruminal acidosis in dairy veterinary practice

Controlling rumen disorders is critical to ensure successful dairy herd health management. Lactation diets of dairy cows are commonly rich in concentrates and low in physically effective fibre. Feeding of these diets increases the risk of rumen disorders with far‐reaching consequences for cattle health, welfare and sustainability of dairy production. The term subacute ruminal acidosis or SARA is often used as a synonym for poor rumen health. Being subclinical, SARA lacks of clear symptoms and is therefore difficult to diagnose and to control in the practice. This review article summarises common and identifies new direct and indirect cow signals related to SARA. We have performed a scientific evaluation and interpretation of each of these cow signals by highlighting their advantages and disadvantages from the practitioner's point of view. The gold standard of SARA cow signals still remains direct measurement of ruminal pH. However, continuous pH monitoring is cost‐intensive and often biased by sensor drift. Single‐point ruminal pH measurements by oral stomach tubing or rumenocentesis have strong limitations. Therefore, there is a need for reliable and robust markers of SARA that are easily accessible and inexpensive. Such indirect parameters are the observation of chewing and feeding activities, as well as the monitoring of milk, faecal, urine and blood variables. Also, novel technologies that allow rapid and non‐invasive measurement of the rumen mucosa thickness and ruminal motility patterns might provide advantages in SARA diagnosis. Due to several constraints of these indirect diagnostic tools, such as limited specificity and sensitivity, we strongly recommend using a combination of the signals to reliably identify cows at risk of SARA in a dairy herd.     

Gene expression profiling of peripheral mononuclear cells in lame dairy cows with foot lesions

Lameness is a major health issue and likely the single most common cause of pain and discomfort in dairy cattle. Appropriate treatment is delayed or neglected due, in part, to lack of reliable detection. Assessment of cows with lameness is currently limited to subjective visual scoring systems based on locomotion and posture abnormalities. These systems are unreliable to detect lameness, and therefore, a large number of cows remain undiagnosed. The objective of this research was to search for potential biomarkers for lameness-associated painful inflammatory foot lesions in dairy cattle using microarray-based gene expression profiling of peripheral blood mononuclear cells (PBMC). BOTL5 microarrays spotted in duplicate with cDNA representing bovine immune response genes were interrogated with cDNA samples in an eight-array, balanced complete block design with dye swap. Samples from eight lame cows with inflammatory foot lesions and from eight sound cows were pair-matched by age, weight, days in lactation, and pregnancy status at time of PBMC collection and directly compared with each other on individual arrays. Statistical analysis of resulting fluorescence intensity data revealed 31 genes that were putatively differentially expressed in lame versus sound cows (P < 0.05). Of these, BLASTn analysis and gene ontology information showed that 28 genes had high similarity or homology to known human and/or rodent genes. Validation of 15 of these genes known to be important in inflammation and pain was carried out using relative quantitative real-time RT-PCR, which confirmed the up-regulation of interleukin (IL)-2 (12.68 ± 1.47-fold increase) and IL-10 (2.39 ± 0.55-fold increase), matrix metalloproteinase-13 (MMP-13) (10.44 ± 1.14-fold increase), and chemokine C–C motif receptor-5 (CCR5) (5.26 ± 1.05-fold increase), in lame relative to sound cows (P ≤ 0.05). Similarly, granulocyte-macrophage colony-stimulating factor receptor alpha chain precursor (GM-CSF-R-alpha) (2.30 ± 0.63-fold increase) and IL-4 (2.06 ± 0.59-fold increase) showed a tendency (P = 0.10) for up-regulation in lame compared to sound cows. PBMC co-expression of IL-2, MMP-13, CCR5 and IL-10, and potentially IL-4 and GM-CSF-R-alpha appears to be a promising, objective sign of lameness-related inflammatory foot lesions in dairy cattle. In conclusion, this study revealed potential biomarkers of the presence of foot lesions that could boost diagnostic accuracy of lameness and, ultimately, help identify animals in need of pain relief.     

Detection of bovine herpesvirus type 4 antibodies and bovine lymphotropic herpesvirus in New Zealand dairy cows

AIM: To detect the presence of bovine herpesvirus (BoHV) type 4 in New Zealand dairy cows with clinical metritis.

METHODS: Serum samples taken from 92 dairy cows with clinical metritis, each from a different farm, were tested for the presence of antibodies against BoHV-4 using a commercially available, indirect ELISA. Peripheral blood mononuclear cells (PBMC) were collected from 10 BoHV-4 seropositive cows, and PBMC were examined by a pan-herpesvirus nested PCR to detect herpesvirus. PCR products were sequenced directly and a proportion of the PCR products were cloned and sequenced to identify the virus present.

RESULTS: Antibodies to BoHV-4 were detected in 23/92 (25%) serum samples. The pan-herpesvirus PCR was positive in 8/10 PBMC samples. Cloning and sequencing identified that all of the eight PCR-positive PBMC contained bovine lymphotropic herpesvirus (BLHV); no BoHV-4 DNA was detected.

CONCLUSIONS: This study reports the finding of the presence of apparent antibodies to BoHV-4, and BLHV DNA in New Zealand dairy cows affected by metritis.

CLINICAL RELEVANCE: Bovine herpesvirus type 4 and BLHV are reported to have the potential to cause reproduction failure in cows. This is the first report of apparent BoHV-4 antibodies, and BLHV in New Zealand. The importance and epidemiology of these viruses in cattle in New Zealand requires further investigation.     

Genetic parameters for total lactation yields of milk,fat, protein,and somatic cell score in New Zealand dairy goats

The aim of this study was to estimate genetic parameters for lactation yields of milk (MY), fat (FY), protein (PY), and somatic cell score (SCS) of New Zealand dairy goats. The analysis used 64,604 lactation records from 23,583 does, kidding between 2004 and 2017, distributed in 21 flocks and representing 915 bucks. Estimates of genetic and residual (co) variances, heritabilities, and repeatabilities were obtained using a multiple‐trait repeatability animal model. The model included the fixed effects of contemporary group (does kidding in the same flock and year), age of the doe (in years), and as covariates, kidding day, proportion of Alpine, Nubian, Toggenburg, and “unknown” breeds (Saanen was used as the base breed), and heterosis. Random effects included additive animal genetic and doe permanent environmental effects. Estimates of heritabilities were 0.25 for MY, 0.24 for FY, 0.24 for PY, and 0.21 for SCS. The phenotypic correlations between MY, FY, and PY ranged from 0.90 to 0.96, and the genetic correlations ranged from 0.81 to 0.93. These results indicate lactation yield traits exhibit useful heritable variation and that multiple trait selection for these traits could improve milk revenue produced from successive generations of New Zealand dairy goats.     

Factors associated with colostrum quality in individual cows from dairy herds in the Waikato region of New Zealand

AIMS: To examine associations between various cow-level factors and quality of first-milking colostrum (measured as Brix), and to evaluate herd-level associations between vaccination against calf diarrhoea and colostrum quality, in cows from dairy herds in the Waikato region of New Zealand.

METHODS: A single colostrum sample was collected, by complete udder evacuation, from each of 20 cows from 29 dairy herds in the Waikato region of New Zealand during the 2016 spring calving period. Vaccination pre-partum with a calf diarrhoea vaccine was used in 15 herds. Each colostrum sample was tested using a digital Brix refractometer. The body condition score of each cow was recorded at the time of sample collection and farmers provided records of clinical mastitis and facial eczema from the previous 12 months, as well as the age and breed of cows. Associations between cow-level variables in non-vaccinated herds and Brix were examined using a multivariable linear mixed model and estimated marginal means obtained for different categories.

RESULTS: Mean Brix of 281 samples from cows in non-vaccinated herds was 18.7 (SD 0.26)%; 63/281 (22.4%) samples had Brix ≥22% and 152/281 (54.1%) had Brix ≥18%. Mean Brix of colostrum samples from cows aged ≥6 years (20.2 (95% CI=19.1–21.2)%) was higher than for samples from 2-year-old cows (18.6 (95% CI=17.3–19.9)%) (p=0.005). Colostrum that was collected at the first milking on the day of calving had higher Brix (20.0 (95% CI=19.1–20.9)%) than colostrum collected from cows that calved the previous day (17.5 (95% CI=16.5–18.4)%) (p<0.001). Mean Brix of colostrum samples from cows which produced ≥8?L (18.2 (95% CI=17.1–19.2)%) tended to be lower than from cows which produced <8?L first-milking colostrum (19.1 (95% CI=18.3–20.0)%) (p=0.08). Among vaccinating herds, 9/15 (60%) had ≥60% colostrum samples with Brix ≥18% compared with 4/14 (29%) of non-vaccinating herds (p=0.04).

CONCLUSIONS AND CLINICAL RELEVANCE: Colostrum quality, as measured by Brix, was associated with the total volume of first-milking colostrum, interval from calving to colostrum collection and cow age. Vaccination against calf diarrhoea was associated with a higher proportion of colostrum samples with adequate Brix. Careful selection of colostrum donor cows should ensure newborn calves are fed adequate quality colostrum which should be beneficial in preventing failure of passive transfer of IgG. Testing of colostrum from individual cows with a Brix refractometer is advocated for the selection of colostrum for feeding newborn calves.     

Accuracy of fetal age estimates using transrectal ultrasonography for predicting calving dates in dairy cows in seasonally calving herds in New Zealand

AIM: To describe the accuracy of transrectal ultrasonography for predicting calving dates in dairy cows under typical New Zealand conditions and to assess potential risk factors for differences between predicted and actual calving dates.

METHODS: Data were collected from 116 seasonally calving herds over 2 years in a retrospective single cohort study. Transrectal ultrasonography was undertaken by experienced veterinarians (n=12) to determine if cows were pregnant, and if so to estimate fetal age. Predicted calving date was calculated by adding 282 days to the estimated conception date. Accuracy was assessed using differences between predicted and actual calving dates for each animal. Potential risk factors for animals calving >10 days before or after their predicted calving date were assessed using multinomial logistic regression models.

RESULTS: The study population comprised 83,104 cows over the 2 years of the study; 75,037 (90.3%) cows calved within 10 days of their predicted calving date, 3,683 (4.4%) calved >10 days earlier, and 4,384 (5.3%) >10 days later, than predicted. Risk factors for calving >10 days before or after the predicted calving date included having >1 artificial insemination (AI) before pregnancy diagnosis (p=0.03), where the cow’s most recent AI was <21 days before the end of the herd’s AI period (p<0.01), and where the diagnosis was made at the second or third herd-visit (p<0.01). The probability of calving being >10 days later than predicted also increased when the fetus was ≥13 weeks old at pregnancy diagnosis (p<0.01).

CONCLUSIONS AND CLINICAL RELEVANCE: In this study, >90% of cows diagnosed pregnant by veterinarians using transrectal ultrasonography calved within 10 days of the predicted calving date. In herds where herd reproductive performance is high, it would be expected that more cows would conceive to their first AI, and potentially fewer cows would have AI close to the end of the herd’s AI period, which would increase diagnostic accuracy. Where herd managers rely on accurate predicted calving dates they should be informed about realistic expected accuracy. For greatest accuracy, a complete AI history should be made available to the person performing the pregnancy diagnoses and cows at most risk of having inaccurate predicted calving dates should be identified.     

Economics of a dry-cow anthelmintic drenching programme for dairy cows in New Zealand. Part 1. Overall response in 47 herds

Trials were conducted on 47 seasonal supply dairy farms (greater than 5500 cows) to assess the economics of a dry-cow anthelmintic drenching programme. The programme was administered during the autumn/winter of 1983 and consisted of two treatments, the first shortly after drying off (late April-early June) and the second shortly before calving (mid-July-late August). The effects of the programme on the body condition of cows over winter and their subsequent milk production and reproductive performance were assessed. Overall, the treatment did not result in a worthwhile improvement in cow condition over winter. However, there was a small, but significant overall increase in milk production (2.24 kg milkfat/cow/lactation = 51.5 l milk; P less than 0.01). Young cows (3 years old) did not respond significantly better than mature cows, but high quality cows (as assessed on the basis of the previous season's milk production) responded significantly better than those of poorer quality. The pre-calving condition of cows did not significantly influence the magnitude of their response. Calving data for the year following the trials indicated that conception rates and time of conception were unaffected by the drench programme. At prices prevailing at the time of the trials, the programme proved to be only marginally economic overall (approximately 1 kg milkfat/cow was required simply to cover the cost of the anthelmintic used). However, levels of response in the individual herds involved varied considerably.     

西北肉用绵羊新品群陶滩寒组合的微卫星标记研究

利用微卫星标记技术,分析了无角陶赛特羊、滩羊、小尾寒羊及其杂交后代滩寒F1、陶滩寒F1 5个绵羊群体的遗传多样性,结果表明,7个微卫星位点均为高度多态位点,平均多态信息含量（PIC）达0.7077～0.8395;5个群体平均位点杂合度达0.7454～0.8665。以Nei氏遗传距离的UPGMA和NJ聚类结果表明,滩羊与滩寒F1具有较近的亲缘关系,小尾寒羊与陶滩寒F1具有较近的亲缘关系,可聚为一类;滩羊、小尾寒羊与陶赛特羊具有较远的亲缘关系。     

Economics of a dry-cow anthelmintic drenching programme for dairy cows in New Zealand. Part 2. Influence of management factors and other herd characteristics on the level of response

The results of a series of trials conducted on 47 seasonal supply dairy farms in the southern North Island of New Zealand, which indicated a statistically significant overall production response of 2.24 kg milkfat/cow/lactation following a 2-treatment dry-cow anthelmintic drenching programme (Bisset et al., 1987), were subjected to further analyses in an attempt to identify any management factors or herd characteristics which may have influenced the levels of production response to treatment in individual herds. The management factors examined included calf drenching practices, types of grazing system, mean pre-calving condition of the herds, and 'nutritional status' of the herds over the milking season. The influence of herd quality, herd size and geographical location were also examined. Considered separately, only calf drenching practice had a significant influence on the level of herd response to treatment. Increases in milkfat production/cow due to the treatment programme were significantly greater (P less than 0.05, t-test) on farms where calves had received a minimal number of drench treatments (less than or equal to 2) between weaning and the end of March (mid-autumn) (+5.32 kg/cow/lactation, P less than 0.01) (= 108.2 l milk), than on farms where calves had received regular 3-4 weekly treatments over the same period (+0.42 kg/cow/lactation, N.S.) While none of the other factors, considered in isolation, appeared to significantly influence the production response of herds, observations on possible interactive effects suggested that the grazing system employed was probably of importance in so far as it determined the level of exposure of cows to calf-contaminated pasture. Thus, the greatest mean response to the treatment programme occurred in herds where cows were overwintered on areas grazed during the milking season by calves which had received less than or equal to 2 drench treatments from weaning until the end of March (+5.95 kg milkfat/cow/lactation, P less than 0.01).     

A review of diagnostic tests for diagnosing failure of transfer of passive immunity in dairy calves in New Zealand

ABSTRACT

The aim of this review is to critically assess the test characteristics and practicality of published data on direct and indirect tests for diagnosing failure of transfer of passive immunity (FPT) in dairy calves in New Zealand, to provide recommendations for veterinary practitioners, and to examine the recommended sample size for assessing herd-level prevalence of FPT and the confidence in the results obtained. The definition of FPT is based on measurement of concentrations of IgG in serum of neonatal calves after colostrum intake. The gold standard method for measurement of concentrations of IgG is radial immunodiffusion. However its cost, requirements for laboratory equipment, and the time taken to obtain results have meant that alternative tests have been developed. The turbidimetric immunoassay and ELISA also directly measure concentrations of IgG. Indirect tests include measurement of concentrations of total proteins (TP) in the laboratory or using a refractometer, γ-glutamyl transferase (GGT) activity, and the zinc sulfate turbidity (ZST) test. Of the indirect tests, measurement of concentrations of TP in the laboratory or using a refractometer combine high specificity and sensitivity with a consistent association with concentrations of IgG in calves between 1–7 days of age. Using a refractometer is less accurate than direct measurement in a laboratory, but is still a suitable test if low cost and speed are important. Although GGT activity is strongly associated with concentrations of IgG in serum, the relationship varies with time after birth. Therefore the target thresholds change with time, increasing error compared to the measurement of concentrations of TP in serum. Similarly, factors other than total concentrations of IgG have a significant effect on the association with ZST test, complicating interpretation. Thus, when direct measurement of concentrations of IgG is not feasible, the recommendation is that concentrations of TP in serum should be used as the diagnostic test for diagnosis of FPT, providing calves are not dehydrated. Using a sample size of 12 calves is suitable for estimating whether the herd-level prevalence of FPT is <20% or >20%, if there are no calves or >5 calves diagnosed with FPT, respectively, but is limited in diagnostic confidence when 1–4 calves test positive. Diagnostic interpretation can be significantly improved if tests of FPT are used alongside information on the likely risk of FPT on the tested farm.