A stochastic model for transmission, extinction and outbreak of Escherichia coli O157:H7 in cattle as affected by ambient temperature and cleaning practices.

Many infectious agents transmitting through a contaminated environment are able to persist in the environment depending on the temperature and sanitation determined rates of their replication and clearance, respectively. There is a need to elucidate the effect of these factors on the infection transmission dynamics in terms of infection outbreaks and extinction while accounting for the random nature of the process. Also, it is important to distinguish between the true and apparent extinction, where the former means pathogen extinction in both the host and the environment while the latter means extinction only in the host population. This study proposes a stochastic-differential equation model as an approximation to a Markov jump process model, using Escherichia coli O157:H7 in cattle as a model system. In the model, the host population infection dynamics are described using the standard susceptible-infected-susceptible framework, and the E. coli O157:H7 population in the environment is represented by an additional variable. The backward Kolmogorov equations that determine the probability distribution and the expectation of the first passage time are provided in a general setting. The outbreak and apparent extinction of infection are investigated by numerically solving the Kolmogorov equations for the probability density function of the associated process and the expectation of the associated stopping time. The results provide insight into E. coli O157:H7 transmission and apparent extinction, and suggest ways for controlling the spread of infection in a cattle herd. Specifically, this study highlights the importance of ambient temperature and sanitation, especially during summer.

Pregnancy risk in beef and dairy cows after supplementing semen with transforming growth factor beta-1 at the time of artificial insemination.

Our objective was to determine if the addition of a concentrated human recombinant transforming growth factor beta-1 (TGF) to bovine semen at the time of AI would result in increased risk of pregnancy in beef and dairy cows. Suckled beef cows (n = 1,132) in 11 herds across 2 states and lactating dairy cows (n = 2,208) in one organic-certified herd were enrolled. Beef cows received fixed-time AI (FTAI) following a 7 d CO-Synch + controlled internal drug release estrous synchronization protocol. Dairy cows were inseminated following observation of natural estrus expression. Cows received either no treatment as a control (CON) or 10 ng of TGF in 10 µL added through the cut-end of a thawed straw of semen immediately prior to AI. At the time of FTAI of beef cows, the mean ±â€…SD age was 5.0 ±â€…2.4 yr, BCS was 5.3 ±â€…0.7, and days postpartum was 78.2 ±â€…15.5 d. The overall pregnancy risk (PR) in beef cows was 55.2% to AI and 90.5% season-long. PR in beef cows was not affected (P = 0.27) by the addition of TGF (53.1% vs. 58.1%). Furthermore, there was no difference (P = 0.88) for season-long PR in beef cows that received TGF (91.2% vs. 91.5%). At the time of insemination of dairy cows, the mean ±â€…SD lactation was 3.0 ±â€…1.3 lactations, BCS was 2.9 ±â€…0.3, days in milk was 115.6 ±â€…56.6 d, and cows had received 2.4 ±â€…1.5 inseminations/cow. The overall pregnancy risk to AI in dairy cows was 23.1%. PR to AI for dairy cows was not affected (P = 0.32) by addition of TGF (22.0% vs. 23.8%). In conclusion, PR to AI was not affected by addition of TGF to thawed semen immediately prior to AI in beef or dairy cows.

Seminal plasma is the fluid portion of the ejaculate that is routinely removed or significantly diluted when preparing semen for artificial insemination. Seminal plasma has been shown to elicit changes to the tissues of the uterus at the time of insemination that improves pregnancy outcomes in rodents and swine. Here, we supplemented the molecule of seminal plasma, transforming growth factor beta-1, to semen at the time of artificial insemination in an attempt to improve pregnancy rates in beef and dairy cattle. In total, 3,340 cows were inseminated; half received no treatment, and the other half received a supplementation of transforming growth factor beta-1. We found that supplementing transforming growth factor beta-1 did not improve the pregnancy rate in beef or dairy cattle. We conclude that the pregnancy rate was not affected by the supplementation of transforming growth factor beta-1 to semen at the time of insemination. Future studies should consider the effects of transforming growth factor beta-1 on other pregnancy outcomes, such as calving rate, birth weight, and postnatal growth.

Associations of automated body condition scores at dry-off and through early lactation with milk yield of Holstein cows.

The objective of this study was to analyze the associations of body condition score (BCS) and BCS change (∆BCS) during the dry period and the first 100 d of lactation with daily milk yield. Examining the involvement of health status in the associations between BCS and milk yield was a secondary objective of this research. Data included 12,042 lactations in 7,626 Holstein cows calving between April 2019 and January 2022 in a commercial dairy operation located in Colorado, USA. BCSs were generated daily by an automated BCS camera system located at the exit of the milking parlor. The assessment points selected for this study were dry-off (BCSdry), calving (BCS1), 7 DIM (BCS7), 14 DIM (BCS14), 21 DIM (BCS21), and nadir (nBCS; defined as the lowest daily BCS from calving to 100 DIM). Subsequently, these BCS were categorized considering quartiles (Q1 = 25% lowest BCS; Q4 = 25% greatest BCS), separately for primiparous and multiparous cows. Changes in BCS were calculated from dry-off to calving (multiparous); and from calving to 7 DIM, 14 DIM, 21 DIM, and nadir and assigned into quartile categories considering Q1 as the 25% of cows with the greatest decrease of BCS. Lactations were classified based on the number of health events before nadir as healthy, affected by one event, or having multiple events. Data were examined in primiparous and multiparous cows separately using ANOVA. The least square means for daily milk at 60 DIM and 305 DIM were calculated by category of BCS and ∆BCS at multiple time points and time periods. Subsequently, lactation curves were created by BCS and ∆BCS categories and by health status. Multivariable models included calving season and BCS1 as covariables. The largest differences in milk yield among categories of BCS and ∆BCS were identified for BCS originated at nadir and for the ∆BCS between calving and nadir. The differences in average daily milk yield between cows in the lowest and the greatest nBCS category (Q1 vs. Q4) were 3.3 kg/d (60 DIM) and 3.4 kg/d (305 DIM) for primiparous cows and 2.4 kg/d (60 DIM) and 2.1 kg/d (305 DIM) for multiparous cows. During the period from calving to nadir, primiparous cows in Q1 (greatest decrease of BCS) produced 4.3 kg/d (60 DIM) and 3.8 kg/d (305 DIM) more than cows in Q4. For multiparous cows, the differences were 3.0 kg/d (60 DIM) and 1.9 kg/d (305 DIM) in favor of Q1 cows. Overall, the associations between BCS and ∆BCS categories and milk yield were not consistent across time and they depended on the parity category. Nonetheless, as the assessment of BCS and ∆BCS approached the nadir, the association between greater milk yield and lower BCS or greater reduction in BCS became more evident.

The objective of this study was to analyze the associations of body condition score (BCS) and BCS change (∆BCS) during the dry period and the first 100 d of lactation with daily milk yield. The involvement of health status in these associations was also evaluated. The relationship between BCS and ∆BCS with milk yield was more evident for nadir BCS (nBCS; defined as the lowest daily BCS from calving to 100 d in milk [DIM]) and for the ∆BCS occurring between calving and nBCS. Cows categorized in the lowest nBCS category and cows with greater loss in BCS from calving to nadir had the greatest average milk yields at 60 DIM and 305 DIM. In agreement, clear patterns for lactation curves were identified when the study cows were categorized by nBCS and ∆BCS for the calving to nBCS period. There was no evident association of health status with the reported relationships between BCS and milk yield.

Factors associated with the time and magnitude of the nadir body condition score in early lactation and its subsequent effects on fertility of Holstein cows.

The objective of this study was to identify variables associated with the time and magnitude of nadir BCS (nBCS) during early lactation and to characterize the effects of these two measurements on the subsequent pregnancy at first artificial insemination (P/AI1) and pregnancy loss of Holstein cows. A retrospective observational study was completed using data collected from 12,042 lactations in 7,626 Holstein cows calving between April 2019 and January 2022 in a commercial dairy operation located in Colorado, USA. Scores generated by an automated BCS camera system at dry-off (BCSdry), calving (BCS1), nadir (nBCS; defined as the lowest daily BCS from calving to 100 DIM), and 21 d post nadir were selected for the analyses and subsequently categorized into quartiles (Q1 = lower nBCS). Changes in BCS were calculated from dry-off to calving (multiparous cows); from calving to nadir; and from nadir to 21 d post nadir and assigned into quartile categories considering Q1 as the 25% of cows with greater loss (pre nadir) or smaller gain (post nadir) of BCS. Data were examined using logistic regression, ANOVA, and time to event analyses. Initial univariable models were followed by multivariable models that considered parity category, calving season, BCS1, and average milk yield up to 30 DIM (M30) as covariables. Sick cows and high producing cows had increased DIM at nadir, while lower BCSdry, greater M30, and occurrence of disease were associated with lower nBCS. Cows that conceived at AI1 had their nBCS earlier (45.1 ± 0.32 d) than cows that remained open after first AI (46.4 ± 0.35 d). On the contrary, DIM at nBCS did not differ by pregnancy loss status. The logistic regression analyses reported smaller odds of P/AI1 in cows in the lower nBCS categories. Likewise, cows with larger loss in BCS between calving and nadir had smaller odds of P/AI1. Although cows that lost their pregnancy had lower nBCS, this association was not consistent across the analyses. The time to and magnitude of nBCS, together with the ∆BCS from calving to nadir could be useful measures to evaluate the transition and early lactation status of dairy cows.

The transition from gestation to lactation requires adjustments in nutrient consumption that are challenging for the postpartum dairy cow. As milk energy output increases more rapidly than energy intake, most cows mobilize fat and labile protein from body energy reserves, which results in reductions in body condition score during early lactation. The objective of this study was to identify variables associated with the extent of the lossin body condition score (BCS) during early lactation and to characterize the effects of the nadir BCS on the subsequent pregnancy at first artificial insemination and pregnancy loss in Holstein cows. Parity, disease presentation, and milk yield were associated with the time and magnitude of nadir BCS. Sick cows and high producing cows had increased DIM at nadir, while lower BCS at dry-off, greater milk yield, and occurrence of disease were associated with lower nadir BCS. Overall, cows with later and lower nadir BCS and greater loss of BCS between calving and nadir had poorer fertility. Monitoring nadir BCS could be a useful tool for evaluating the transition and early lactation status of dairy cows.

Genome-wide association and genomic prediction for a reproductive index summarizing fertility outcomes in U.S. Holsteins.

Subfertility represents one major challenge to enhancing dairy production and efficiency. Herein, we use a reproductive index (RI) expressing the predicted probability of pregnancy following artificial insemination (AI) with Illumina 778K genotypes to perform single and multi-locus genome-wide association analyses (GWAA) on 2,448 geographically diverse U.S. Holstein cows and produce genomic heritability estimates. Moreover, we use genomic best linear unbiased prediction (GBLUP) to investigate the potential utility of the RI by performing genomic predictions with cross validation. Notably, genomic heritability estimates for the U.S. Holstein RI were moderate (h2 = 0.1654 ± 0.0317-0.2550 ± 0.0348), while single and multi-locus GWAA revealed overlapping quantitative trait loci (QTL) on BTA6 and BTA29, including the known QTL for the daughter pregnancy rate (DPR) and cow conception rate (CCR). Multi-locus GWAA revealed seven additional QTL, including one on BTA7 (60 Mb) which is adjacent to a known heifer conception rate (HCR) QTL (59 Mb). Positional candidate genes for the detected QTL included male and female fertility loci (i.e. spermatogenesis and oogenesis), meiotic and mitotic regulators, and genes associated with immune response, milk yield, enhanced pregnancy rates, and the reproductive longevity pathway. Based on the proportion of the phenotypic variance explained (PVE), all detected QTL (n = 13; P ≤ 5e - 05) were estimated to have moderate (1.0% < PVE ≤ 2.0%) or small effects (PVE ≤ 1.0%) on the predicted probability of pregnancy. Genomic prediction using GBLUP with cross validation (k = 3) produced mean predictive abilities (0.1692-0.2301) and mean genomic prediction accuracies (0.4119-0.4557) that were similar to bovine health and production traits previously investigated.

Association between body condition score fluctuations and pregnancy loss in Holstein cows.

The objective of this study was to characterize the associations between body condition score (BCS) and BCS change (∆BCS), determined by an automated camera system during early lactation and close to artificial insemination (AI), and the subsequent pregnancy loss (PL) in Holstein cows. A secondary objective was to determine the impact of disease on PL, considering multiple time periods relative to AI. Data from 9,430 lactations in 6,884 Holstein cows in a commercial dairy operation located in Colorado, USA were included in this retrospective observational study. Cows were subject to first AI at about 80 DIM (primiparous) and 60 DIM (multiparous), following a double OvSynch protocol. Pregnancy diagnosis was performed via transrectal ultrasonography on day 32 ± 3 after AI and reconfirmed on day 80 ± 3 after AI. Cameras mounted on the sorting gate at each exit (n = 2) of the milking parlor generated BCS on a five-point scale with 0.1 increments. The BCS at calving (BCS1), 21 DIM (BCS21), 56 DIM (BCS56), AI resulting in pregnancy (BCSAI), and 90 d post AI (BCSAI90) were selected for the analyses and subsequently categorized as low (≤ lower quartile), moderate (interquartile range), and high (≥ upper quartile). Changes in BCS were calculated by periods of interest as change from calving to 21 DIM; change from calving to 56 DIM; change from 56 DIM to AI; and change from AI to 90 d post AI and assigned into categories to facilitate the analysis. Data were examined using logistic regression, considering parity category, season at calving and AI, DIM at AI, milk yield up to 60 DIM, and occurrence of disease as covariables. The logistic regression analyses indicated that the odds of PL were greater in cows in the low BCS category relative to cows in the high BCS category at 56 DIM (OR 95% CI = 1.41 [1.12-1.79]), AI (1.31 [1.05-1.65]), and 90 d post AI (1.38 [1.10-1.74]). Likewise, cows with large loss in BCS between calving and 21 DIM (1.46 [1.10-1.94]) and loss in BCS between AI and 90 d post AI (1.44 [1.15-1.81]) had greater odds of PL compared with cows with no loss of BCS within the same period. Occurrence of disease at all the time periods considered in the analysis had a consistent detrimental impact on maintenance of the pregnancy, supporting the concept that pre and postconceptional disease affects embryonic survival. Overall, low BCS, more pronounced reductions in BCS occurring closer to AI, and occurrence of disease resulted in greater PL in this Holstein population.

Maintenance of the pregnancy until term is a key factor for adequate fertility in dairy operations, as early embryonic mortality in lactating dairy cows is significant. The objective of this study was to investigate the association between the changes in body condition scores (BCS) during early lactation and in the proximity of artificial insemination (AI) and embryo mortality in Holstein cows. A secondary objective was to determine the impact of disease on pregnancy loss (PL), considering multiple time periods relative to AI. Our study population included 9,430 lactations in 6,884 Holstein cows that had their daily BCS determined by an automated camera system during early lactation and close to AI. Cows were diagnosed pregnant via transrectal ultrasonography on day 32 after AI and reconfirmed on day 80 after AI. Overall, the dynamics of BCS differed between cows that lost or did not lose their pregnancy. Low BCS and more pronounced reductions in BCS occurring closer to the AI, as well as concurrent disease, resulted in greater levels of PL.

Evaluation of Contamination in Milk Samples Pooled From Independently Collected Quarters Within a Laboratory Setting.

The primary objective of this observational study was to evaluate the prevalence of contamination from independently collected quarter-level milk samples pooled in a laboratory and subjected to bacterial culture. To address this objective, weekly quarter-level milk samples were collected longitudinally from a cohort of 503 primiparous cows from five organic dairy farms during the first 5 weeks after calving. Individual quarter milk samples were pooled in a laboratory using aseptic technique ("lab-pooled") and subjected to bacterial culture. In the sample set of 2,006 lab-pooled milk samples, 207 (10.3%) were classified as contaminated using a standard definition (i.e., growth of three or more distinct microorganisms). Subsequent culturing of corresponding quarter-level milk samples revealed that many of the contaminated lab-pooled sample results (i.e., 46.7%) were the result of intramammary infections with different pathogens across the quarters, rather than actual contamination within any single quarter (i.e., "true contamination"). The odds of true contamination were lower when the lab-pooled sample exhibited growth of three microorganisms compared to more than 3 microorganisms. Our findings suggest that pooling of quarter samples within a laboratory setting may yield lower rates of contamination compared to those previously reported from samples composited on-farm, but that current cut-offs to define contamination may need to be evaluated for use with lab-pooled samples. Further investigation of use of lab-pooled samples may be warranted to reduce costs while still providing useful scientific insight.

Current attitudes of veterinarians and producers regarding the use of local and systemic analgesia in beef and dairy cattle in the United States.

OBJECTIVE: To survey cattle producers and veterinarians about the use of analgesia on US cattle operations. SAMPLE: 1,187 members of the following database, electronic mailing lists, and social media groups: FarmProgress master file, American Association of Bovine Practitioners, Academy of Veterinary Consultants, National Milk Producers Federation Farm Evaluators, Dairy Moms Facebook group, and Dairy Girl Network Facebook group. PROCEDURES: An online survey was developed to gather information about the frequency of local and systemic analgesia use for common painful procedures and diseases in cattle < 2, 2 to 12, and > 12 months old. Respondents also rated their extent of agreement with each of 10 statements related to pain management in cattle. The survey was available from June 11 to August 10, 2018. Descriptive data were generated. Logistic regression was used for comparisons among cattle age groups and respondents on the basis of their industry role. RESULTS: In general, frequency of analgesia use increased as cattle age increased, regardless of the procedure or disease. The odds of analgesia use were lower for men, compared with women, and greater for veterinarians, compared with producers. Many respondents indicated they were cognizant of the benefits of analgesia use in cattle but perceived federal regulations and drug costs as impediments to the implementation of pain mitigation protocols on cattle operations. CONCLUSIONS AND CLINICAL RELEVANCE: Results provided insight into current perceptions and use of analgesia in cattle, which can be used to guide implementation of pain mitigation protocols on US beef and dairy cattle operations.

Effect of peripartal administration of mycobacterium cell wall fraction on health and fertility of Holstein cows under organic-certified management.

The main objective of this study was to evaluate the effect of peripartal administration of a commercially available nonspecific immune stimulant (mycobacterium cell wall fraction; MCWF [Amplimune, NovaVive Inc., Napanee, ON, Canada]) on the incidence of disease during early lactation and subsequent fertility of dairy cows. A second objective was to characterize the dynamics of circulating white blood cells (WBC) and metabolic markers following treatment administration. Cows in an United States Department of Agriculture (USDA) organic-certified dairy herd were blocked by parity and, based on sequential calving dates, randomly assigned to receive two injections (5 mL s.c.) of either a placebo (saline solution) as a control (CON; n = 71) or MCWF (n = 65) at enrollment (7 d before expected calving) and within 24 h after calving. Blood samples were collected from a subsample of the study population (MCWF = 16; CON = 18) for WBC count at enrollment, at day 2 post enrollment, and at days 1, 3, 7, and 14 after calving. Serum fatty acids, beta-hydroxybutyrate, and Ca concentrations were determined at days 1 and 7 postpartum (MCWF = 21; CON = 21). Main outcome variables included incidence risk of peripartal and early lactation health disorders and pregnancy at first artificial insemination (AI), at 100, and at 150 days in milk (DIM). In addition, the average daily milk yield up to 90 DIM and death and live culling before 305 DIM were compared. Treatment effects were assessed using multivariable logistic regression, time-to-event analyses, and repeated measures analysis of variance (ANOVA). A treatment effect on the incidence risk of some of the health disorders in the study was established. Incidence risk of metritis and clinical mastitis <28 DIM was smaller in MCWF than in CON cows (36.9% vs. 50.7% and 6.3% vs. 19.7%, respectively). On the contrary, the incidence risk of respiratory disease <28 DIM was smaller in CON (0%) than in MCWF (7.7%). Reproductive performance of multiparous cows was affected by MCWF administration: pregnancy at first AI and pregnancy at 100 and 150 DIM were greater in MCWF than in CON (35.6% vs. 19.2%; 51.1% vs. 25.0%; and 64.4% vs. 40.4%, respectively). Overall, median intervals from calving to pregnancy were 90 vs. 121 d in MCWF and CON cows, respectively. No treatment effects on the dynamics of circulating WBC or in postpartum metabolic status were established. No differences for milk yield or for the proportion of cows that survived up to 305 DIM were determined, although cows in MCWF left the herd earlier than cows in CON. In conclusion, incidence risks of metritis and mastitis in early lactation were smaller in cows receiving MCWF, whereas the incidence risk of respiratory disease was smaller in CON. Fertility significantly improved in MCWF compared with CON cows. As this study was performed in an organic-certified dairy, specific health and reproductive management practices may affect the external validity of the current findings.

Association between CARD15/NOD2 gene polymorphisms and paratuberculosis infection in cattle.

Paratuberculosis represents a major problem in farmed ruminants and at the present is considered a potential zoonosis. The disease is caused by Mycobacterium avium subsp. paratuberculosis, and susceptibility to infection is suspected to have a genetic component. Caspase recruitment domain 15 (CARD15) gene encodes for a cytosolic protein implicated in bacterial recognition during innate immunity. Crohn's disease (CD) is an idiopathic inflammatory bowel disease in humans comparable in many features to bovine paratuberculosis involving an abnormal mucosal immune response. The association between mutations in the CARD15 gene and increased risk of Crohn's disease has been described. The objective of this candidate gene case-control study was to characterize the distribution of three polymorphisms in the bovine CARD15 gene and test their association with paratuberculosis infection in cattle. Three previously reported single nucleotide polymorphisms (E2[-32] intron 1; 2197/C733R and 3020/Q1007L) were screened for the study population (431 adult cows). The statistical analysis resulted in significant differences in allelic frequencies between cases and controls for SNP2197/C733R (P<0.001), indicating a significant association between infection and variant allele. In the analysis of genotypes, a significant association was also found between SNP2197/C733R and infection status (P<0.0001); cows with the heterozygous genotype were 3.35 times more likely to be infected than cows with the reference genotype (P=0.01). Results suggest a role for CARD15 gene in the susceptibility of cattle to paratuberculosis infection. These data contribute to the understanding of paratuberculosis, suggest new similarities with Crohn's disease and provide new information for the control of bovine paratuberculosis.

Relationships among quarter milk leukocyte proportions and cow and quarter-level variables under different intramammary infection statuses.

The use of milk leukocyte differential (MLD) test has been proposed as a complement to somatic cell count (SCC) to assess the presence and the severity of intramammary infection. However, detailed information regarding the behavior of MLD under different physiological or pathological stages of the cow is nonexistent. The objective was to analyze the association between milk leukocyte proportions provided by a commercial automated MLD test and multiple cow and quarter-level variables. The study population consisted of 104 Holstein cows (32 primiparous and 72 multiparous) in one farm. Cows were categorized by days in milk as early (<50 DIM; n=29), middle (50-250 DIM; n=25), and late lactation (>250 DIM; n = 50). Milk from 416 quarters was collected and analyzed for lymphocytes (LYM), neutrophils (NEU), and macrophages (MAC) counts using an automated milk fluorescence microscopy system. Concurrently, a sterile composite milk sample was collected from each cow for pathogen identification through microbiological culture. Culture results were classified as no growth (NOG), gram-negative (GN), gram-positive (GP), or other (OTH). Milk leukocyte proportions varied depending on the level of total leukocyte counts (TLC; P < 0.001). Similarly, leukocyte ratios (NEU:LYM, NEU:MAC, and phagocyte:LYM) were different for multiple TLC categories (P < 0.05). There was no association between parity number and MLD; however, cows in early lactation had the greatest proportions of NEU and LYM. Leukocyte ratios varied depending on parity number and stage of lactation. Cows in the medium milk-yield category had the smallest proportions of NEU and LYM, and there was significant variation in leukocyte ratios, depending on the level of milk yield. In healthy quarters, MLD were not associated with quarter position; however, the NEU:MAC ratio was greater in rear quarters than in front quarters. In quarters with TLC >100,000, NEU% was greater in rear quarters than in front quarters (P = 0.03). For quarters with pathogen growth, TLC was greatest for GN followed by OTH and GP (P < 0.001). Milk LD depended on the isolated pathogen group, although the magnitudes of the differences were small. Although the changes in the proportions of leukocytes in milk were associated with categories of TLC, levels of milk yield, and mastitis-causing pathogen groups, the deviations were small in magnitude. Additional research is necessary to determine the potential applications for this methodology.

Evaluation of milk components as diagnostic indicators for rumen indigestion in dairy cows.

OBJECTIVE To identify milk component alterations that might be useful for detecting cows with rumen indigestion. DESIGN Prospective case-control study. ANIMALS 23 Holstein cows with rumen indigestion (cases) and 33 healthy cohorts (controls) from 1 herd. PROCEDURES Cases were defined as cows between 30 and 300 days postpartum with a > 10% decrease in milk yield for 2 consecutive milkings or > 20% decrease in milk yield from the 10-day rolling mean during any milking, abnormally decreased rumen motility, and no other abnormalities. Each case was matched with 2 healthy cows (controls) on the basis of pen, parity, days postpartum, and mean milk yield. Some cows were controls for multiple cases. All cows underwent a physical examination and collection of a rumen fluid sample for pH measurement at study enrollment. Individual-cow milk yield and milk component data were obtained for the 16 milkings before and after study enrollment. Rumen motility and pH and milk components were compared between cases and controls. RESULTS Rumen motility for cases was decreased from that of controls. Cases had an abrupt increase in milk fat percentage and the milk fat-to-lactose ratio during the 2 milkings immediately before diagnosis of rumen indigestion. Receiver operating characteristic analyses revealed that a 10% increase in the milk fat-to-lactose ratio had the highest combined sensitivity (57%) and specificity (85%) for identifying cows with rumen indigestion. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that a positive deviation in the milk fat-to-lactose ratio might be useful for identifying cows with rumen indigestion.

Lymphatic fluid for the detection of Mycobacterium avium subsp. paratuberculosis in cows by PCR, compared to fecal sampling and detection of antibodies in blood and milk.

Johne's disease (JD), caused by Mycobacterium avium subsp. paratuberculosis (MAP), can cause considerable economic losses in affected herds. Early diagnosis of JD is hampered by the chronic nature of the disease with a slow subclincal progression. The aim of the present study was to challenge the hypothesis that lymphatic fluid is of diagnostic value in the early stages of the disease. Lymphatic fluid from 122 animals was collected and tested for MAP by nested PCR for IS900 and compared to the results of testing for MAP in feces (culture), blood and milk (ELISA) in 110 of these samples. MAP was detected by PCR in 27.1% of the lymph samples. Agreement between the tests was poor: 6.9% of the lymph positive cows were also positive in all other tests applied, and 69.0% had negative results in fecal culture, blood and milk ELISA. Resampling of 25 cows after 8 to 12 and 16 to 20 months revealed 20.0% lymph positive animals at the first, 5.5% at the second and 27.8% at the third sampling, respectively. Only one cow showed positive lymph-PCR results at more than one sampling date. Lymph-positive cows had a 7.2 times greater likelihood of being culled within 8 to 12 months after sampling, compared to negative cows, mainly due to other health issues than JD. It can be concluded, that lymphatic fluid might be promising for the detection of early MAP-infection in cows, but further studies to elucidate the potential of this diagnostic approach are needed.

Association of hematologic variables and castration status at the time of arrival at a research facility with the risk of bovine respiratory disease in beef calves.

OBJECTIVE: To determine the association of CBC variables and castration status at the time of arrival at a research facility with the risk of development of bovine respiratory disease (BRD). DESIGN: Retrospective cohort study. ANIMALS: 1,179 crossbred beef bull (n = 588) and steer (591) calves included in 4 experiments at 2 University of Arkansas research facilities. PROCEDURES: Calves underwent processing and treatments in accordance with the experiment in which they were enrolled. Castration status and values of CBC variables were determined at the time of arrival at the facilities. Calves were monitored to detect signs of BRD during a 42-day period. RESULTS: The areas under the receiving operator characteristic curves for CBC variables with significant contrast test results ranged from 0.51 (neutrophil count) to 0.67 (eosinophil count), indicating they were limited predictors of BRD in calves. The only CBC variables that had significant associations with BRD in calves as determined via multivariable logistic regression analysis were eosinophil and RBC counts. The odds of BRD for bulls were 3.32 times the odds of BRD for steers. CONCLUSIONS AND CLINICAL RELEVANCE: Results of this study indicated that low eosinophil and high RBC counts in blood samples may be useful for identification of calves with a high risk for development of BRD. Further research may be warranted to validate these variables for prediction of BRD in calves. Calves that were bulls at the time of arrival had a higher risk of BRD, versus calves that were steers at that time.

The collection of lymphatic fluid from the bovine udder and its use for the detection of Mycobacterium avium subsp. paratuberculosis in the cow.

The objective of the current study was to evaluate the feasibility of lymph collection from the bovine udder and to investigate if the lymphatic fluid might be of diagnostic value in cows infected with Mycobacterium avium subsp. paratuberculosis (MAP), the etiologic agent of paratuberculosis. Lymph fluid collection was attempted from 58 cows, and the reactions of the cows as well as the level of difficulty of the procedure were recorded in 56 animals. Lymph samples (51 in total) were tested for the presence of MAP by nested polymerase chain reaction. Collection of the lymphatic fluid caused no or mild signs of discomfort in 94.6% of the cows; in 51.8% of cows, lymphatic fluid was attained on the first attempt, while sample collection was unsuccessful in 12.1%. Mycobacterium avium subsp. paratuberculosis was detected in 43.1% of all lymph samples. The bacterium was present in 66.7% of cows with clinical Johne's disease, in 42.8% of asymptomatic cows with a positive or suspicious enzyme-linked immunosorbent assay (ELISA) result in blood, and in 38.7% of cows with a negative ELISA result in blood. The present study shows that the procedure was well tolerated by most cows and can easily be performed on farm. The current report of the isolation of MAP from lymph fluid suggests that the present approach could be used for the early detection of Johne's disease in cattle.

Risk haplotype analysis for bovine paratuberculosis.

Paratuberculosis (Johne's disease), caused by Mycobacterium avium subsp. paratuberculosis, is an important disease for bovines, although its genetic basis is poorly understood. In this study, three candidate genes were typed to study the associations between single nucleotide polymorphisms (SNPs) and paratuberculosis susceptibility (measured in a 1 or 0 form) at the haplotype level. A significant risk haplotype, constructed by a variant allele (C) at the first SNP and a common allele (A) at the second SNP, within the CARD15 gene was detected to trigger genetic effects on paratuberculosis infection in an overdominace manner. Marginally significant haplotypes were identified for the other two genes. The results obtained will provide scientific guidance about the selection and prediction of resistance types in bovines.

Candidate gene polymorphisms (BoIFNG, TLR4, SLC11A1) as risk factors for paratuberculosis infection in cattle.

Paratuberculosis (Johne's disease) imposes a significant problem to the world dairy and beef industries and today is considered a potential zoonosis. The disease is caused by Mycobacterium avium subsp. paratuberculosis and is characterized by progressive weight loss and profuse diarrhoea. Susceptibility to infection is suspected to have a genetic component, and moderated values for heritability of infection have been reported. Interferon gamma is an inducible cytokine with a crucial role in the innate host response to intracellular bacteria. Toll-like receptors are trans-membrane structures responsible for coordination of innate and adaptive immune responses. The solute carrier family 11 member 1 (SLC11A1, formerly NRAMP1) gene plays an important role in innate immunity, preventing bacterial growth in macrophages during the initial stages of infection. The objective of this candidate gene case-control study was to characterize the distribution of polymorphisms in three candidate genes related to the immune function; interferon gamma (BoIFNG), toll-like receptor 4 (TLR4), and SLC11A1 genes and to test their role as potential risk factors for paratuberculosis infection in cattle. The statistical analysis demonstrated significant differences in allelic frequencies between cases and controls for BoIFNG-SNP(1)2781 and SLC11A1 microsatellites, indicating a significant association between infection and variant alleles. In the analysis of genotypes, a significant association was also found between infection status and BoIFNG-SNP(1)2781 and SLC11A1-275-279-281 microsatellites. However, when variables such as breed and age were included in the multivariate logistic regression analysis, a tendency toward statistical significance for the effect of polymorphisms in the odds of infection was only found for alleles SLC11A1-275 and 279.