Prophylactic use of decoquinate for infections with Cryptosporidium parvum in experimentally challenged neonatal calves

OBJECTIVE: To evaluate the effect of daily oral administration of decoquinate to neonatal calves experimentally challenged with various numbers of Cryptosporidium parvum oocysts. DESIGN: Clinical trial. ANIMALS: 75 calves. PROCEDURE: Calves were purchased from a commercial dairy during a 5-week period. Calves were housed in individual hutches and fed milk replacer with or without decoquinate (2 mg/kg [0.9 mg/lb per day]). Calves were randomly assigned to treatment and 1 of 5 challenge groups (0, 50, 100, 1000, or 10,000 C. parvum oocysts in 60 mL of saline [0.9% NaCl] solution administered p.o. on the day after arrival). Calves were maintained in the study for as long as 28 days. Calves were clinically assessed for diarrhea and dehydration. Fecal samples were submitted for oocyst enumeration 3 times each week. RESULTS: Treatment did not affect number of days to first watery feces (diarrhea), number of days to first oocyst shedding, or duration of diarrhea or oocyst shedding. Duration of oocyst shedding was significantly associated with challenge dose of oocysts administered to calves and number of days to first oocyst shedding. Duration of diarrhea and number of days to first oocyst shedding were significantly associated with week of arrival and number of days to first watery diarrhea. CONCLUSIONS AND CLINICAL RELEVANCE: Daily treatment with decoquinate at the dosage used in this study did not affect oocyst shedding or clinical signs associated with cryptosporidiosis. However, there was an indication that if the number of oocysts calves received could be reduced, then the duration of oocyst shedding and, hence, environmental loading of C. parvum oocysts could be reduced.     

Duration of naturally acquired giardiosis and cryptosporidiosis in dairy calves and their association with diarrhea

OBJECTIVE: To determine duration of infection and association of infection with diarrhea for dairy calves with naturally acquired cryptosporidiosis and giardiosis. DESIGN: Cohort study. ANIMALS: 20 Holstein calves on a single dairy farm. PROCEDURE: Fecal samples were collected 3 times/wk for the first 45 days after birth, then weekly until calves were 120 days old and examined for Giardia duodenalis cysts and Cryptosporidium parvum oocysts. Calves were monitored for diarrhea during the first 45 days after birth; during each episode of diarrhea, fecal samples were examined for parasitic, bacterial, and viral pathogens. RESULTS: All 20 calves shed Giardia cysts and Cryptosporidium oocysts at some time during the study. Mean ages at which Giardia cysts and Cryptosporidium oocysts were first detected were 31.5 and 16.3 days, respectively. Mean number of Giardia cysts in feces remained high throughout the study, whereas Cryptosporidium occysts decreased to low or undetectable numbers 2 weeks after infection. Eighteen calves had a total of 38 episodes of diarrhea during the first 45 days after birth. Giardia duodenalis was the only pathogen identified during 6 (16%) episodes, C parvum was the only pathogen identified during 9 (24%) episodes, and G duodenalis and C parvum were identified together during 10 (26%) episodes. CONCLUSIONS: Prevalences of giardiosis and cryptosporidiosis were high in these calves, and both parasites were associated with development of diarrhea. Cryptosporidium parvum was an important pathogen when calves were < 1 month old, but G duodenalis was more important when calves were older. Calves cleared C parvum infections within 2 weeks; however, G duodenalis infections became chronic in these calves.     

Transmission of bovine coronavirus and serologic responses in feedlot calves under field conditions

OBJECTIVE: To compare shedding patterns and serologic responses to bovine coronavirus (BCV) in feedlot calves shipped from a single ranch in New Mexico (NM calves) versus calves assembled from local sale barns in Arkansas (AR calves) and to evaluate the role of BCV on disease and performance. ANIMALS: 103 feedlot calves from New Mexico and 100 from Arkansas. PROCEDURES: Calves were studied from before shipping to 35 days after arrival at the feedlot. Nasal swab specimens, fecal samples, and serum samples were obtained before shipping, at arrival, and periodically thereafter. Bovine coronavirus antigen and antibodies were detected by use of an ELISA. RESULTS: NM calves had a high geometric mean titer for BCV antibody at arrival (GMT, 1,928); only 2% shed BCV in nasal secretions and 1% in feces. In contrast, AR calves had low antibody titers against BCV at arrival (GMT, 102) and 64% shed BCV in nasal secretions and 65% in feces. Detection of BCV in nasal secretions preceded detection in feces before shipping AR calves, but at arrival, 73% of AR calves were shedding BCV in nasal secretions and feces. Bovine coronavirus infection was significantly associated with respiratory tract disease and decreased growth performance in AR calves. CONCLUSIONS AND CLINICAL RELEVANCE: Replication and shedding of BCV may start in the upper respiratory tract and spread to the gastrointestinal tract. Vaccination of calves against BCV before shipping to feedlots may provide protection against BCV infection and its effects with other pathogens in the induction of respiratory tract disease.     

Natural and experimental infection of neonatal calves with Clostridium difficile

Clostridium difficile toxins were associated with calf diarrhea in a recent retrospective study; however, no causal relationship has been prospectively investigated. This infection study tested whether the oral inoculation of neonatal calves with a toxigenic strain of C. difficile (PCR-ribotype 077) results in enteric disease. Fourteen 6-24 h old male colostrums-fed Holstein calves, received either three doses of C. difficile (1.4 x 10(8) +/- 3.5 x 10(8) cfu) (n = 8) or sterile culture broth (n = 6). Calves were euthanized on day 6 or after the onset of diarrhea, whichever came first. Fecal and intestinal samples were blindly cultured for C. difficile, and tested for its toxin A/B (C. difficile TOX A/B II ELISA, Techlab). PCR-ribotyping was used to compare inoculated and recovered isolates. Diarrhea was observed in all control calves and 3/8 of inoculated calves (p = 0.03), but it did not occur in calves that tested positive for C. difficile toxins. Fecal toxins were identified only from two controls. PCR-ribotyping confirmed the presence of C. difficile PCR-ribotype 077 in samples of all inoculated calves, but not from controls. The identification of five other PCR-ribotypes in 3/8 (37.5%) and 2/6 (33.3%) of inoculated and control calves, respectively, indicated early natural infection (< or = 24h of age). Five of 14 cecal samples had C. difficile (p = 0.01). In conclusion, the oral administration of C. difficile PCR-ribotype 077 to neonatal calves resulted in fecal/intestinal colonization but not in detection of toxins, or signs of enteric disease. Further studies are required to investigate the clinical relevance of C. difficile in calves.     

A case-control study of selected pathogens including verocytotoxigenic Escherichia coli in calf diarrhea on an Ontario veal farm.

A case-control study of diarrheal disease in veal calves was conducted over a three month period on a single large veal farm in southern Ontario. One hundred diarrheic calves (cases) were identified by visual examination of their feces. Each case was matched to two nondiarrhetic controls from the same room on the same day, and a fecal sample was obtained from each animal. Fecal consistency of cases and controls was observed daily for one week following sample collection. Control calves which developed diarrhea during that period were excluded from the study. Breed, sex and the date and nature of antimicrobial drugs administered to each calf were recorded. Moisture content of fecal samples was measured by weighing samples before and after oven drying. Samples were screened for verocytotoxigenic Escherichia coli (VTEC) using a Vero cell assay, for enterotoxigenic E. coli (ETEC) using an immunoblot procedure with anti-K99 monoclonal antibodies, and for Salmonella species using modified semi-solid Rappaport-Vassiliadis medium. A latex agglutination test was used to detect rotaviruses, and samples were examined for cryptosporidia using sucrose wet mounts. No VTEC were identified in cases or controls. One calf was positive for Salmonella and three were positive for ETEC. Rotaviruses were detected in four cases and four controls. A significant positive association was found between diarrhea and infection with Cryptosporidium. This study thus provided no evidence of an association between diarrhea and infection with either VTEC, ETEC, Salmonella spp. or rotaviruses in the population examined. On the other hand our results do suggest that Cryptosporidium infection may promote transient diarrheal disease in veal calves in Ontario.     

Calf-level risk factors for neonatal diarrhea and shedding of Cryptosporidium parvum in Ontario dairy calves

This work was conducted to investigate calf-level factors that influence the risk of neonatal diarrhea and shedding of Cryptosporidium parvum oocysts in calves, on dairy farms in Ontario with histories of calf diarrhea or cryptosporidiosis. Fecal samples were collected weekly for 4 weeks from each of 1045 calves under 30 days of age on 11 dairy farms in south-western Ontario during the summer of 2003 and the winter of 2004. A questionnaire designed to gather information on calf-level management factors was administered on farm for each calf in the study. Samples were examined for C. parvum oocysts by microscopy, and a subset of specimens was also tested for enterotoxigenic Escherichia coli, Salmonella, bovine rotavirus and bovine coronavirus. The consistency of each sample was scored and recorded at the time of collection in order to assess the presence or absence of diarrhea. In addition, a blood sample was taken from each calf upon enrollment in the study, for assessment of maternal antibody transfer and for polymerase chain reaction testing for persistent bovine viral diarrhea virus infection. Using the GLLAMM function in Stata 9.0, multilevel regression techniques were employed to investigate associations between management practices and the risk of C. parvum shedding or diarrhea. C. parvum oocysts were detected in the feces of 78% of the 919 calves from which all four fecal samples had been collected. Furthermore, 73% of the 846 calves for which all four fecal consistency scores had been recorded were diarrheic at the time of collection of at least one sample. Significant predictors of the calf-level risk of C. parvum shedding included the use of calf diarrhea prophylaxis in pregnant cows, and the type of maternity facilities in which the calves were born. Factors associated with an increased risk of diarrhea were leaving the calf with the dam for more than an hour after birth, and the birth of a calf in the summer as opposed to winter. Calves shedding C. parvum oocysts had 5.3 (95% CI 4.4, 6.4) times the odds of diarrhea than non-shedding calves, controlling for other factors included in the final multivariable model. Furthermore, infected calves shedding more than 2.2 x 10(5) oocysts per gram of feces were more likely to scour than infected calves shedding lower numbers of oocysts (OR= 6.1, 95% CI 4.8, 7.8). The odds of diarrhea in calves shedding oocysts that had been allowed to remain with their dams for more than an hour were higher than the odds of diarrhea in shedding calves that had been separated from their dams within an hour after birth.     

A longitudinal study of bovine coronavirus enteric and respiratory infections in dairy calves in two herds in Ohio

This prospective longitudinal study examined the epidemiology and disease syndrome associated with bovine coronavirus (BCV) infections in a cohort of 8 conventional calves from 0 to 120 days of age, in two dairy herds in Ohio. The periods of respiratory shedding of BCV were determined by direct immunofluorescent (DIF) staining of nasal epithelial cells and ELISA of nasal swab supernatant fluids. The periods of fecal shedding of BCV were determined by ELISA and immunoelectron microscopy (IEM). The isotype-specific antibody titers to BCV in serum (at selected intervals between 0 and 120 days of age) and the post-suckling (24 to 48 h after birth) total immunoglobulin levels were examined by ELISA and zinc sulfate turbidity tests, respectively. Of the 8 calves studied, 4 had evidence of BCV respiratory (by DIF or ELISA) or enteric infections (by IEM or ELISA) in association with diarrhea or rhinitis, even though 7 of 8 calves showed increases in one or more serum antibody isotypes to BCV and 6 of 8 calves showed BCV respiratory or enteric antigen shedding by ELISA. Serological antibody titer increases occurred in 3 calves before 30 days of age and in 4 calves after 30 days of age; two of the latter calves had a second rise in serum antibody titers to BCV after the initial rise. A serological antibody titer increase was not observed in one calf. This suggests that BCV infections may be very common in a closed herd and may occur in older calves, although many may be subclinical and some may be recurrent. There were no statistically significant correlations between total serum immunoglobulin levels or BCV antibody isotype titers in serum (24-48 h after birth) and clinical disease or infection by BCV; however, calves with low levels of IgA BCV antibodies in serum (24-48 h after birth) had a significantly greater average number of days with diarrhea than those calves having high levels of IgA BCV-specific antibodies in serum.     

Epidemiology of Clostridium difficile on a veal farm: prevalence, molecular characterization and tetracycline resistance

Concern has been raised about the potential for Clostridium difficile to be a bovine and foodborne pathogen, yet limited study has been performed in cattle, and none in veal calves. This study evaluated the epidemiology and microbiology of C. difficile on one veal farm. Rectal swabs were obtained from calves within 48 h of arrival and at one, 17 and 21 weeks later. Selective culture for C. difficile was performed. Isolates were characterized by PCR ribotyping and PCR for tcdA, tcdB and cdtA. Tetracycline resistance and resistance genes were investigated. Multivariable logistic regression models were constructed to determine the relationship between shedding of the bacterium and specific ribotypes and the independent variables: time of sampling and area of housing. Calves were twice more likely to test positive 1 week after arrival (51%) when compared to initial results (32%). Shedding at 17 and 21 weeks was significantly lower (2% at both samplings). Ribotype 078 was the most common. Twelve different ribotypes were present initially with only three ribotypes found subsequently. Seventy-six percent (40/53) of isolates initially recovered were tetracycline resistant compared to 93% (81/87) from 2nd sampling. Tetracycline resistance genes were detected in 24% (13/53) of isolates during 1st and in 55% (50/91) during 2nd sampling. The high prevalence of pathogenic C. difficile in veal calves could be of zoonotic concern. The low prevalence before slaughter may be of importance for the evaluation of foodborne risks. Oxytetracycline administration to calves may have an impact on prevalence of C. difficile colonization.     

Epidemiologic factors and isotype-specific antibody responses in serum and mucosal secretions of dairy calves with bovine coronavirus respiratory tract and enteric tract infections.

Blood, feces, and nasal swabs specimens were collected 12 to 24 hours after birth and then 3 times/week (blood only once per week) from one group of 10 calves until they were 10 weeks old and from a second group of 10 calves until they were 10 to 20 weeks old. Colostrum was collected from all calves' dams and tears from 5 randomly selected calves in the first group. All fecal and nasal specimens were assayed for bovine coronavirus (BCV) antigens by ELISA. Nasal epithelial cells were examined for BCV antigens by direct immunofluorescence. Isotype antibody titers to BCV in all samples from 5 calves in group 1 were evaluated by ELISA. Zinc sulfate turbidity (ZST) values were determined on the first serum samples taken from all calves in group 1. To determine whether any correlation existed between ZST values, isotype antibody titers to BCV (12 to 24 hours after birth), number of respiratory sick days, number of enteric sick days, or days to first shedding of virus, a Spearman rank order correlation coefficient was done. Bovine coronavirus respiratory tract and enteric tract infections were common on this farm. Most initial infections developed when calves were 1 to 3 weeks old; however, there were also multiple incidences of shedding of viral antigens or seroconversions at later times during the study. Persistence of infection or reinfection of the upper respiratory tract with BCV was common. Colostral antibody titers to BCV (IgG1) were in all cows at moderate amounts; however, calf serum antibody titers and ZST values (12 to 24 hours after birth) were highly variable.(ABSTRACT TRUNCATED AT 250 WORDS)     

牛病毒性腹泻病毒BVDV-JL株的分离与鉴定

本研究从吉林某牛场表现严重腹泻症状濒死牛的胸腺病料样品中分离一株病毒,该病毒在MDBK细胞中盲传4代无细胞病变产生,而通过RT-PCR和间接免疫荧光试验、微量血清中和试验检测表明该分离病毒株为牛病毒性腹泻病毒(BVDV),并命名为BVDV-JL.将BVDV-JL株F4代细胞培养液(10<'7.13>TCID<,50>/...     

Development of nasal, fecal and serum isotype-specific antibodies in calves challenged with bovine coronavirus or rotavirus

Unsuckled specific pathogen free calves were inoculated at 3-4 weeks of age, either intranasally (IN) or orally (O) with bovine coronavirus or O plus IN (O/IN) or O with bovine rotavirus. Shedding of virus in nasal or fecal samples, and virus-infected nasal epithelial cells were detected using immunofluorescent staining (IF), ELISA or immune electron microscopy (IEM). Isotype-specific antibody titers in sera, nasal and fecal samples were determined by ELISA. Calves inoculated with coronavirus shed virus in feces and virus was detected in nasal epithelial cells. Nasal shedding persisted longer in IN-inoculated calves than in O-inoculated calves and longer than fecal shedding in both IN and O-inoculated calves. Diarrhea occurred in all calves, but there were no signs of respiratory disease. Calves inoculated with rotavirus had similar patterns of diarrhea and fecal shedding, but generally of shorter duration than in coronavirus-inoculated calves. No nasal shedding of rotavirus was detected. Peak IgM antibody responses, in most calves, were detected in fecal and nasal speciments at 7-10 days post-exposure (DPE), preceeding peak IgA responses which occurred at 10-14 DPE. The nasal antibody responses occurred in all virus-inoculated calves even in the absence of nasal shedding of virus in rotavirus-inoculated calves. Calves inoculated with coronavirus had higher titers of IgM and IgA antibodies in fecal and nasal samples than rotavirus-inoculated calves. In most inoculated calves, maximal titers of IgM or IgA antibodies correlated with the cessation of fecal or nasal virus shedding. A similar sequence of appearance of IgM and IgA antibodies occurred in serum, but IgA antibodies persisted for a shorter period than in fecal or nasal samples. Serum IgG1 antibody responses generally preceeded IgG2 responses and were predominant in most calves after 14-21 DPE.     

Evaluation of concurrent shedding of bovine coronavirus via the respiratory tract and enteric route in feedlot cattle

OBJECTIVE: To assess the relationship between shedding of bovine coronavirus (BCV) via the respiratory tract and enteric routes and the association with weight gain in feedlot cattle. ANIMALS: 56 crossbred steers. PROCEDURES: Paired fecal samples and nasal swab specimens were obtained and were tested for BCV, using antigen-capture ELISA. Paired serum samples obtained were tested for antibodies to BCV, using antibody-detection ELISA. Information was collected on weight gain, clinical signs, and treatments for enteric and respiratory tract disease during the study period. RESULTS: Number of samples positive for bovine respiratory coronavirus (BRCV) or bovine enteric coro navirus (BECV) was 37/224 (17%) and 48/223 (22%), respectively. Some cattle (25/46, 45%) shed BECV and BRCV. There were 25/29 (86%) cattle positive for BECV that shed BRCV, but only 1/27 (4%) cattle negative to BECV shed BRCV. Twenty-seven of 48 (56%) paired nasal swab specimens and fecal samples positive for BECV were positive for BRCV. In contrast, only 10/175 (6%) paired nasal swab specimens and fecal samples negative for BECV were positive for BRCV. Only shedding of BECV was associated with significantly reduced weight gain. Seroconversion to BCV during the 21 days after arrival was detected in 95% of the cattle tested. CONCLUSIONS AND CLINICAL IMPLICATIONS: Feedlot cattle infected with BCV after transport shed BCV from the respiratory tract and in the feces. Fecal shedding of BCV was associated with significantly reduced weight gain. Developing appropriate control measures for BCV infections could help reduce the decreased weight gain observed among infected feedlot cattle.     

Influence of diet on the shedding of Candida glabrata by experimentally infected preweaned calves

The influence of three diets, comprising dam's milk (DM) from the same farm, commercial milk replacer with (CMRL) or without 3.2% lactose (CMR), on the duration and intensity of Candida glabrata shedding in the faeces of preweaned calves following experimental oral infection was examined. Shedding of other potential enteric pathogens was also monitored. The duration and intensity of C. glabrata shedding in DM-fed calves were reduced significantly compared with the calves fed the two diets based on milk replacers. Consequently, feeding calves with DM might disrupt the infective cycle, resulting in the yeast's elimination from a farm. In the CMR and CMRL groups, the periods of intensive shedding of C. glabrata and rotavirus overlapped but no diarrhoea was associated with the shedding of either microorganism. There was no evidence that lactose diminished colonization in vivo.     

Evaluation of lasalocid and decoquinate against coccidiosis resulting from natural exposure in weaned dairy calves

Eighteen female Holstein calves, raised as natural herd additions under conditions typical of a well-managed midwestern United States dairy farm, were used in a natural-exposure study to determine the anticoccidial efficacies of lasalocid and decoquinate. Calves were allotted to 6 treatment blocks of 3 calves each as they were weaned. Within each block, calves were randomly assigned to be given either lasalocid or decoquinate or to remain as a nonmedicated control. Calves were given medication for 90 days and remained separated from other calves for 120 days. Adjusted weight gains were consistently greater in calves that were given medication; however, differences were not statistically significant. Fecal specimens were obtained from calves at weekly intervals during the study. Overall, oocyst shedding was low. During the medication period, quantitative mean fecal shedding of oocysts was reduced eightfold in calves given decoquinate and four-fold in calves given lasalocid, as compared with nonmedicated control calves. During the period following the medication period, calves that had been controls shed fewer oocysts than did calves that had previously been given medication. A pairwise comparison of the proportion of specimens that were oocyst-positive was made to assess qualitative oocyst shedding among treatment groups. During the medication period, qualitative oocyst shedding (all species, Eimeria bovis, E zuernii, species other than E bovis and E zuernii) was greater in controls than in either lasalocid-or decoquinate-treated groups. Like-wise, lasalocid-medicated calves shed oocysts more frequently than did the decoquinate-medicated group. After medication, qualitative findings were reversed. Little diarrhea was noticed in treatment or control calves during the study.(ABSTRACT TRUNCATED AT 250 WORDS)