Risk of infection with Cryptosporidium parvum and Cryptosporidium hominis in dairy cattle in the New York City watershed

OBJECTIVE: To determine the risk posed by Cryptosporidium parvum and Cryptosporidium hominis from dairy cattle in the New York City watershed (NYCW). SAMPLE POPULATION: Samples from cattle at risk for shedding Cryptosporidium organisms on randomly selected dairy farms in the NYCW. PROCEDURE: Feces were collected for 4 years from calves at risk for infection on 37 dairies. Oocysts were detected by use of centrifugation concentration-flotation microscopy. The DNA was directly isolated from fecal samples and used to amplify fragments of the small subunit ribosomal RNA and thrombospondin-related adhesion protein C-2 genes by use of nested polymerase chain reaction assays. Small subunit ribosomal RNA fragments were restriction digested by the enzyme Vspl and thrombospondin-related adhesion protein C-2 fragments were digested by Eco91l to distinguish between C hominis (formerly known as genotype 1) and C parvum (formerly known as genotype 2). RESULTS: Of 437 fecal samples examined, 214 contained oocysts. Amplicons were generated for 200 samples. We can be certain, with 95% confidence, that cattle in the NYCW did not harbor C hominis. CONCLUSIONS AND CLINICAL RELEVANCE: Cryptosporidium infections in cattle are under examination because of the potential contamination of public waters by manure. Although cattle may be the source of zoonotic infection via C parvum, they pose little risk for C hominis (the strain commonly isolated from humans in waterborne outbreaks of disease). Other sources of oocysts should be considered when investigating outbreaks attributable to contaminated urban drinking water because cattle pose only a small risk via shedding of C hominis.     

Incidence of Cryptosporidium parvum in the dairy cattle population in a New York City Watershed

A longitudinal study of 2-year duration was conducted to determine the risk, as measured by incidence rate, of Cryptosporidium parvum infection among dairy cattle in the Catskill/Delaware Watershed of New York City (NYC), and the factors that predispose animals to the likelihood of infection. A proportional sampling scheme with follow up at quarterly farm visits was employed for heifers and cows. Additionally, all calves born on the 39 study farms were sampled once during the first four weeks of life and at least once more before weaning. Samples were analyzed for the presence of C. parvum using a quantitative centrifugation concentration flotation technique and a C. parvum-specific enzyme-linked immunosorbent assay (ELISA). Of the 9914 fecal samples collected, 747 were found to contain C. parvum. The average number of oocysts detected was 1.3x10(5)/g (range: 1.0/g--8.2x10(6)/g). The average age at time of first detection of the organism was 15.0 days with a standard deviation of 6.59 days. The age range of animals infected with C. parvum in the study population was 3--60 days (inclusive). The unadjusted (crude) incidence rate of C. parvum among the entire study population was 2.05 per 1000 animal-days. The unadjusted incidence rate among pre-weaned calves was 15.55 per 1000 animal-days. After controlling for age and prior protozoal risk level, no seasonal impact on the incidence of C. parvum was detected among animals less than 61 days by negative binomial regression. A seasonal impact was identified among the oocyst counts of infected animals after controlling for age and prior protozoal risk level.     

Risk factors associated with Cryptosporidium parvum infection in cattle

A 2-year, cross-sectional study was conducted to identify risk factors for Cryptosporidium sp. infection in bovine farms in central Italy. Faecal samples were collected on 248 farms, from 2024 calves and analysed using ELISA and immunofluorescent assay (IFA) commercial kits. In all 101 samples confirmed to be positive with IFA, the aetiological agent was identified as Cryptosporidium parvumand a large genetic variability was detected by subtype analysis. The prevalence of farm infection ranged from 3.4% to 35.6%. Univariate analysis showed a number of putative risk factors, including the type of farm, stalling of calves, late supply of colostrum, number of heads and contact between calves and adults. However, multivariate analysis confirmed that the higher risk for calves was associated with housing calves separately from their dams, a characteristic practice of dairy herd, whereas calves being nursed by their dams, a characteristic of cow-calf herd resulted as a protective factor.     

Severe respiratory disease in dairy cattle in New York State associated with bovine respiratory syncytial virus infection

Severe respiratory disease associated with bovine respiratory syncytial virus (BRSV) infection has been identified in dairy cattle in New York State. The cases identified occurred in dairy calves and heifers. The disease was characterized in 4 animals by pathologic changes including interstitial pneumonia, necrotizing bronchiolitis with multinucleated syncytial epithelial cells and interstitial emphysema. BRSV antigen was demonstrated in lung samples or was isolated in tissue culture in all 4 cases. A retrospective survey of 6279 bovine diagnostic accessions between 1977 and 1982 revealed 66 cases of interstitial pneumonia, often with concurrent bronchiolitis. In this 5 year period, only 1 case in 1981 had interstitial pneumonia and bronchiolitis with pathologic features consistent with BRSV infection. It is concluded that pathogenic BRSV has entered New York State and that it is contributing to clinical respiratory disease in dairy cattle.     

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.     

Association between management practices and within-herd prevalence of Cryptosporidium parvum shedding on dairy farms in southern Ontario

To identify management practices associated with an increased within-herd prevalence of Cryptosporidium parvum shedding on dairy farms in southern Ontario, fecal samples were taken from 1089 calves aged 7-28 days, from 119 herds. Information on management practices was obtained by administering a questionnaire compiled using a modified Delphi technique. Data were analyzed using univariable and multivariable negative binomial regression. Overall, 30% of the calves in the study were shedding C. parvum oocysts, with at least one positive calf detected in 77% of herds. Within-herd prevalence ranged from 0 to 80%. Predictors significantly associated with an increased prevalence of shedding in multivariable modelling were the use of calf scour prophylaxis in cows (risk ratio [RR] 1.70, P<0.01) and calves (RR 1.38, P=0.02) and the feeding of milk replacer in the first week of life (RR 1.40, P=0.02). In contrast, the presence of concrete flooring in calf housing areas (RR 0.59, P<0.01) and the use of soap or detergent when washing calf feeding utensils (RR 0.61, P<0.01) appeared to be protective.     

Epidemiologic study of Giardia sp. infection in dairy cattle in southeastern New York State

To identify animal and management factors associated with the risk of Giardia sp. infection in dairy cattle in southeastern New York State, an observational analytical epidemiologic study was conducted. A random sample of 2943 animals in 109 dairy herds located in five counties of southeastern New York was selected from the target population. Fecal samples were collected from animals in the study population and examined for the presence of Giardia sp. using a quantitative centrifugation concentration flotation technique. Data on each animal, and on general management, maternity, preweaning, and postweaning practices were collected by personal interview with the farmer or farm manager. The significance of association of these factors, within each management practice group, with the risk of infection was evaluated using the logistic regression analysis. Weighted indices for each of these four groups were developed, and were evaluated together with the age of the animal and season of sampling for their significance of association with the risk of infection using mixed effect logistic regression analysis. Only the maternity management practices, age of the animal, and the season of sampling were significantly associated with the risk of infection with Giardia sp. Summer housing of bred heifers, on pasture or in tie stalls, was associated with increased risk of infection. Calves that were fed fresh colostrum or separated immediately after birth from the dam were at decreased risk of infection. The risk of infection decreased with the age of the animal and varied by the season of sampling. There was no significant extra binomial variation in the risk of infection in this data. If consideration is to be given to reducing the risk of infection with this parasite, management must be a top priority, especially in younger animals.     

Prevalence of Giardia sp. Cryptosporidium parvum and Cryptosporidium andersoni (syn. C. muris) [correction of Cryptosporidium parvum and Cryptosporidium muris (C. andersoni)] in 109 dairy herds in five counties of southeastern New York

A cross-sectional study was undertaken to determine the prevalence of Giardia sp. (G. duodenalis group), Cryptosporidium parvum and Cryptosporidium andersoni (C. muris) [corrected] in dairy cattle in three different age groups, and to evaluate the association of age and season with prevalence. One hundred and nine dairy farms, from a total of 212 farms, in five counties of southeastern New York volunteered to participate. On these farms, 2943 fecal samples were collected from three defined age groups. The farms were randomly assigned for sampling within the four seasons of the year. Each farm was visited once during the study period from March 1993 to June 1994 to collect fecal samples. Demographic data on the study population was collected at the time of sampling by interviewing the farm owner or manager. At collection, fecal samples were scored as diarrheic or non-diarrheic, and each condition was later related to positive or negative infection with these parasites. Fecal samples were processed using a quantitative centrifugation concentration flotation technique and enumerated using bright field and phase contrast microscopy. In this study, the overall population prevalence for Giardia sp. was 8.9%; C. parvum, 0.9%; and C. muris, 1.1%. When considering animals most at the risk of infection (those younger than 6 months of age) Giardia sp. and C. parvum was found in 20.1 and 2.4% of the animals, respectively. Giardia sp. and C. muris were found in all age groups. There was no significant seasonal pattern of infection for any of these parasites.     

Cross-sectional study of faecal shedding of Giardia duodenalis and Cryptosporidium parvum among packstock in the Sierra Nevada Range

Faecal specimens from 305 horses and mules used as packstock at one of 17 commercial or governmental (National Park Service, US Forest Service) operations were examined for Giardia duodenalis and Cryptosporidium parvum using immunofluorescent microscopy. Fourteen packstock (4.6%) were shedding G. duodenalis cysts, with herd-level prevalences ranging 0-22%. Number of packstock in the corral, size of corral and density of packstock in the corral were associated with the odds of shedding G. duodenalis cysts. None of the horses had detectable C. parvum oocysts. Assuming a sensitivity of at least 43% and a specificity of 100% for our assay, the estimated maximum true prevalence of shedding of C. parvum for packstock would be < or = 2.3% of the population. These data suggest that faecal dispersal of C. parvum on back country watersheds is unlikely with packstock.     

Prevalence of Cryptosporidium parvum infection and pattern of oocyst shedding in calves in Japan

Cryptosporidium parvum infection and the pattern of oocyst shedding were observed in calves. A total of 480 fecal samples were collected from 30 calves (age, < or =30 days) over a period of 10 months from June 1998 to March 1999. A sucrose centrifugal flotation technique revealed 28/30 (93%) calves were passing Cryptosporidium oocysts. Oocyst shedding was first detected on the sixth day after birth, with 8% of the calves testing positive. This rate increased day by day and reached approximately 80% by day 15. Oocyst shedding varied from 1 to 13 days, with a mean of 7 days. Calves infected with C. parvum had a significantly higher rate of diarrhea (33%) than non-infected calves (8%) (P<0.05), suggesting C. parvum infection as the likely cause. The mean number of oocysts excreted by calves < or =30 days old was approximately 6x10(7) per gram of feces. These results indicated that one calf would excrete some 6x10(11) oocysts in the first month after birth, taking both the quantity of feces in a day and the period of excretion into consideration. Accordingly, it is clear that calves are important in the spread of cryptosporidiosis to calves and humans.     

Factors associated with seroprevalence of Neospora caninum in dairy cattle in southeastern Brazil

Neosporosis is an infectious disease caused by Neospora caninum, an obligate intracellular cyst-forming protozoan considered a major cause of miscarriage in dairy cattle in many parts of the world. This cross-sectional study evaluated the relationship between reproductive abnormalities and seropositivity to N. caninum in 1,204 dairy cows from 40 farms located in the state of Minas Gerais, southeastern Brazil. Producers were interviewed, and blood samples were collected to perform indirect immunofluorescence tests (IFAT 1:200). Association between reproductive abnormalities and seropositivity in cattle was evaluated with generalized estimating equations. The true herd-level seroprevalence of N. caninum was 95 % (83.3–99.1), while the individual-level true seroprevalence was 21.6 % (19.2–24.2). Several reproductive abnormalities were significantly associated with seropositivity to N. caninum: occurrence of repeated estrus (p?=?0.02; OR?=?3.84; 95 % CI?=?1.239–11.893), repeated miscarriages (p?=?0.001; OR?=?2.54; 95 % CI?=?1.423–5.402), and temporary anestrus (p?=?0.001; OR?=?3.44; 95 % CI?=?1.976–5.994). Furthermore, loose dogs (p?=?0.041; OR?=?2.20; 95 % CI?=?1.033–4.672) when fed raw meat (p?=?0.001; OR?=?1.91; 95 % CI?=?1.443–2.519) are risk factors for N. caninum infection. We observed that seropositivity to N. caninum in cattle increases risk of miscarriage by almost twice throughout the reproductive life of cows (p?=?0.004; OR?=?1.978; 95 % CI?=?1.249–3.131). Serologic evidence in this study indicates that N. caninum infection is widely distributed among dairy herds and significantly associated with reproductive disorders, especially miscarriage, repeated estrus, and temporary anestrus.     

Factors associated with the spread of clinical vesicular stomatitis in California dairy cattle

In 1983, vesicular stomatitis entered the state of California through infected cattle purchased in Idaho and sold to 5 California dairies. This study examined management, environmental, and host factors which were thought to be associated with disease spread. The use of coarse roughage and hard-pelleted concentrates, the presence of uneaten feed potentially contaminated with virus-laden saliva, increased interpen movement of cows, poor ground surface conditions, poor milking hygiene, and poor teat sanitation were management factors observed to be associated with clinical signs. The amount of milk production, age, or days in milk of individual cows seemed to have an important role in the spread of clinical disease. Recommendations are given for control and prevention of clinical signs and, therefore, the severity of disease during epizootics of vesicular stomatitis in California dairies.     

Correlated time to event data: Modeling repeated clinical mastitis data from dairy cattle in New York State

Mastitis is the most prevalent production disease in dairy herds worldwide and is considered to be the most economically important disease of dairy cattle. Modeling the risk of cows contracting mastitis is therefore of great interest for both targeting prevention programs and evaluating treatment protocols. Clinical mastitis (CM) is a disease of recurrent nature, thus correlation between the subsequent events within one cow may be present. This would violate the assumption behind most statistical time-to-event models. In the case of time to event models, the semi-parametric Cox regression models have become the default tool in modeling the time to an event. Limited methods are currently available to evaluate marginal and random (frailty) effects to account for multiple correlation sources. The objective of this study was to explore the implications of using several Cox or related semi-parametric or parametric models to estimate the hazard for CM in the presence of correlation between events. We evaluated the Andersen-Gill model which uses robust standard errors to account for the correlation, the Conditional Anderson-Gill model that uses stratification to account for event dependence, the Frailty model that introduces a random term to account for unobserved (cow level) heterogeneity, and a related generalized linear mixed model that uses Poisson regression to allow multi-level modeling of time-to-event data. We analyzed data on the occurrence of CM from five dairy farms in New York State. Data were from 8206 cows with 721, 275, 119, and 57 first, second, third, and fourth occurrences of CM, respectively, in the same lactation. The analysis of our sample dataset demonstrated that both cow- and farm-level correlation are present in the case of CM. The Conditional Frailty model was able to model one source of correlation in a random effect and one in a fixed effect. Poisson modeling allowed for simultaneous estimation of within cow correlation and within herd correlation.     

The prevalence of Cryptosporidium, and identification of the Cryptosporidium horse genotype in foals in New York State

To date, little is known about the prevalence, genotypes and zoonotic potential of Cryptosporidium spp. affecting horses, especially in North America. A cross-sectional study was conducted in New York, USA between February 25th and May 1st 2009. Fecal samples were collected from three hundred and forty nine 1-10-week-old foals and their dams on 14 different broodmare farms. All fecal samples were screened for Cryptosporidium spp. using a direct immunofluorescence assay (DFA). DNA extraction and PCR-RFLP analysis of the small-subunit (SSU) rRNA gene were performed on all the foal samples. PCR-positive samples were subtyped by DNA sequencing of the 60-kDa glycoprotein (gp60) gene. On DFA, 13/175 (7.4%) foal samples and 3/174 (1.7%) mare samples were designated positive for Cryptosporidium spp., whereas on SSU rRNA-based PCR, 9/175 (5.1%) foal samples were positive. Cryptosporidium PCR-positive foals were significantly older (13-40 days, median age of 28 days) compared with negative foals (4-67 days, median 18 days, p=0.02). The number of foals with diarrhea or soft feces was not significantly different between positive and negative foals (p=0.09). PCR-RFLP analysis of the SSU rRNA gene and DNA sequencing of the gp60 gene identified the parasite as subtype VIaA14G2 of the horse genotype. This is the first report of a group of foals affected with the Cryptosporidium horse genotype, which has recently been detected in humans. As other contemporary molecular studies have identified C. parvum in foals, it seems that equine cryptosporidiosis should be considered a zoonosis.     

Serologic evidence of coronavirus infection in New York and New England dairy cattle with winter dysentery.

Acute and convalescent sera were collected from 8 dairy herds with classic clinical features of winter dysentery. An enzyme-linked immunosorbent assay was used to measure coronavirus antibody titers, employing calf diarrhea coronavirus as antigen. Twenty-two of the 35 animals tested (63%) showed a greater than or equal to 4-fold seroconversion. Adult cattle in all 8 herds seroconverted. These findings complement previously reported immunoperoxidase and electron microscopic evidence, suggesting an etiologic role for an enteric coronavirus in this disease.     

Environmental factors associated with lameness in dairy cattle

An epidemiological survey of the incidence of different foot diseases causing lameness was undertaken on 80 French dairy farms between 1979 and 1980. A total of 29.5 cases of lesions per 100 cows was recorded in that period, which represents a higher incidence that of clinical mastitis (20.37).

Data concerning the incidence of foot lesions, farm structures, herd management system, feeding and soil characteristics were analysed using the ξ² test of independence and one-way analysis of variance. Results showed that a high incidence of infectious and metabolic foot disorders was associated with loose housing, feeding based on maize silage, absence of a footbath, foot trimming and mineral supplementation.

A relationship was also observed between articular lesions and grass-silage feeding. Tied-housed cows fed hay were less affected. Regular hoof trimming at pasture and a long time spent checking the animals were found to have a positive effect on foot health. A low incidence of foot disorders was also associated with a larger variety in dietary components. A high incidence of lameness was reported in large herds kept on rough floors.