Cryptosporidium parvum infection and management-based risk factors of dairy calves in Taiwan

Cryptosporidiosis is one of the major causes of diarrhea in calves. Cryptosporidium parvum is considered the most important calf diarrhea pathogen in the Cryptosporidium species. Not only could infected calves spread C. parvum, but infected adult cattle could also shed oocysts. The objectives of this study were (1) to investigate the prevalence of C. parvum in dairy herds in Taiwan, including calves, the dams in delivery enclosure, the floor, and the drinking water; (2) to clarify the relationship of diarrhea, management, and C. parvum infection. Twenty dairy herds in Taiwan were selected by random sampling, including 226 calves and 198 dams, and other environmental samples were collected. A questionnaire was filled out by the farm owners to collect information regarding the management of calves and the delivery enclosure. Hierarchical logistic regression was used to analyze the risk factors for C. parvum infection. The prevalence of C. parvum infection in calves was 26.5% (60/226), while in dams, it was 19.7% (39/198). The C. parvum infection in calves increased with environmental contamination of C. parvum and clinical signs of diarrhea, while it decreased with a yard provided in the delivery enclosure. In conclusion, the management of the delivery enclosure appears to be more important for preventing cryptosporidiosis in calves in Taiwan.     

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.     

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.     

Prevalence of Cryptosporidium parvum infection in southwestern Ontario and its association with diarrhea in neonatal dairy calves

Cryptosporidium parvum infection was detected in 203 (40.6%) of 500 Ontario dairy calves aged 7 to 21 d, on a convenience sample of 51 farms with a history of calf diarrhea. Within-farm prevalence ranged from 0% to 70%, and both shedding and intensity of shedding were significantly associated with diarrhea. This parasite appears to be common in Ontario dairy calves and important as a cause of dairy calf scours in the province.     

奶牛隐孢子虫病流行病学调查及初生犊牛感染试验

应用饱和糖溶液漂浮法和改良抗酸染色法调查郑州、商丘和洛阳3个地区的5个奶牛场、2个专业村的582份粪样，查出阳性样品64份，隐孢子虫总阳性率11％（64／582），发现两种不同形态的卵囊，根据其形态结构等特点鉴定为小球隐孢子虫（C．parvum）和安氏隐孢子虫（C．andersoni）。其中2个场奶牛感染安氏隐孢子虫，有1个场的奶牛感染小球隐孢子虫，且感染强度较小。犊牛感染率较育成牛、成年牛高。并进行小球隐孢子虫分离株对初生犊牛的致病性试验，其结果为潜隐期7天，排卵囊高峰出现在感染后第16天，高峰期5天。剖检后消化道黏膜经抗酸染色鉴定，仅在回肠中段发现卵囊。     

Prevalence of Cryptosporidium parvum infection in Punjab (India) and its association with diarrhea in neonatal dairy calves

A prevalence study was contemplated to determine the prevalence of Cryptosporidium spp. in dairy farms in Punjab, India. The cryptosporidium oocysts were detected from 50 and 25.68% from 80 diarrheic and 74 non-diarrheic animals, respectively. Both shedding and intensity of shedding were significant in calves with diarrhea. The Cryptosporidium spp. appears to be common in dairy calves and an important contributor of calf diarrhea in the Punjab province. The prevalence of the infection peaked in young calves between 0 and 30 days in both the diarrheic and non-diarrheic groups (86.4 and 66.6%, respectively). The percentage distribution of positive samples, with reference to age groups of diarrheic and non-diarrheic animals was negatively correlated with increase in age. High mortality rate and case fatality rate of 35.2 and 44.4% were observed in young calves between 0 and 30 days of age.     

Number of Cryptosporidium parvum oocysts or Giardia spp cysts shed by dairy calves after natural infection

OBJECTIVE: To determine the total number of Cryptosporidium parvum oocysts and Giardia spp cysts shed by dairy calves during the period when they are most at risk after natural infection. ANIMALS: 478 calves naturally infected with C. parvum and 1,016 calves naturally infected with Giardia spp. PROCEDURE: Oocysts or cysts were enumerated from fecal specimens. Distribution of number of oocysts or cysts versus age was used to determine the best fitting mathematic function. Number of oocysts or cysts per gram of feces for a given duration of shedding was computed by determining the area under the curve. Total number of oocysts or cysts was calculated by taking the product of the resultant and the expected mass of feces. Results: Intensity of Cparvum oocyst shedding was best described by a second-order polynomial function. Shedding increased from 4 days of age, peaked at day 12, and then decreased. An infected 6-day-old calf would produce 3.89 x 10(10) oocysts until 12 days old. Pattern of shedding of Giardia spp cysts was best described by exponential functions. Intensity of shedding increased from 4 days of age, peaked at day 14, and then decreased. An infected calf would produce 3.8 x 10(7) cysts from day 50 until day 56. CONCLUSIONS AND CLINICAL RELEVANCE: The large number of oocysts and cysts shed indicates that shedding by dairy cattle poses a risk for susceptible calves and people. Estimates reported here may be useful to aid in designing cost-effective strategies to manage this risk.     

Antibody fusions reduce onset of experimental Cryptosporidium parvum infection in calves

Cryptosporidium parvum is one of the main causes of diarrhea in neonatal calves resulting in significant morbidity and economic losses for producers worldwide. We have previously demonstrated efficacy of a new class of antimicrobial antibody fusions in a neonatal mouse model for C. parvum infection. Here, we extend efficacy testing of these products to experimental infection in calves, the principal target species. Neonatal calves were challenged with C. parvum oocysts and concomitantly treated with antibody-biocide fusion 4H9-G1-LL37 over the course of four days. This resulted in reduced severity of the disease when compared to control animals. Overall clinical health parameters showed significant improvement in treated animals. Oocyst shedding was reduced in treated when compared to control animals. Control of oocyst shedding is a prerequisite for breaking the cycle of re-infection on dairy farms. Antibody-biocide fusion products thus have the potential to reduce the impact of the infection in both individual animals and in the herd.     

Prevalence of Cryptosporidium infection in dairy calves in western Washington

The prevalence and intensity of Cryptosporidium infection were examined in 445 Holstein calves at 10 dairy farms in western Washington, near Seattle. Fifty-one percent (176) of calves in the 7- to 21-day-old age group (n = 342) were positive for oocysts in the feces by carbolfuchsin staining. Prevalence and intensity of infection were highest in calves 8 to 14 days old; prevalence was 60% in this group, and 48% of the Cryptosporidium-positive calves had oocyst shedding at a 4+ level. A seasonal pattern in prevalence was not evident.     

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.     

Prevalence and risk factors for Cryptosporidium spp. infection in young calves

A cross-sectional study was designed to investigate the prevalence and risk factors for Cryptosporidium infection in young calves. Forty-one farms in a discrete, densely farmed 100 km2 area of North West England were visited over a 3-week period and 215 faecal samples were collected from young calves. Farms were not selected on the basis of existing scour problems. At the time of sampling, several investigator-observed variables were recorded at the pen, animal and stool levels. Samples were screened and 60/215 were confirmed as positive by PCR of the 18S rRNA gene. Risk factors for infection were explored using multilevel multivariable logistic regression with farm as a random effect. Age was significant in the final model, with a higher risk of infection in calves aged 8-21 days, when compared to those aged 0-7 days. The depth of the bedding was also significant in the final model, with calves housed in bedding 11-15 cm deep being at lower risk of infection than those on beds 0-5 cm deep. Consistency of the faeces was highly correlated with age and colour of the faeces and was not significantly associated with infection when these variables, and clustering at farm-level, were accounted for. This is interesting as Cryptosporidium is considered to be a primary enteropathogen. The results suggest that intervention strategies should be targeted at calves under 21 days old. These animals represent a significant reservoir of infection on the farm and may also pose a risk to public health, assuming that the species and genotypes shed are zoonotic pathogens.     

Antibody reactivity to Cryptosporidium parvum in saliva of calves after experimental infection

Antibodies against Cryptosporidium parvum in the saliva and sera of three calves experimentally infected with this parasite were examined by an indirect immunofluorescence antibody test and immunoblotting. Salivary anti-C. parvum IgA antibody appeared on day 12 post-challenge and had a tendency to increase transiently between days 15 and 30 post-challenge. Salivary anti-C. parvum IgG antibody levels showed a gradual increase along with the change in IgA antibody levels during the infection. In contrast, serum anti-C. parvum IgA antibody levels showed only a slight increase between days 15 and 30 post-challenge. Serum anti-C. parvum IgG antibody levels rose on day 12 post-challenge and one calf maintained relatively high level up to the end of the experiment. In immunoblotting, an antigen with a molecular mass of 15 kDa was found to react strongly to salivary IgA antibody and a 27 kDa antigen to react to serum IgG antibody.     

Factors associated with shedding of Cryptosporidium parvum versus Cryptosporidium bovis among dairy cattle in New York State

OBJECTIVE: To isolate and speciate Cryptosporidium DNA from fecal samples obtained from dairy cattle in New York State and identify factors associated with whether cattle were shedding Cryptosporidium parvum versus Cryptosporidium bovis. DESIGN: Cross-sectional study. SAMPLE POPULATION: 115 fecal samples positive for DNA coding for the Cryptosporidium 18S rRNA gene from dairy cattle in New York State. PROCEDURES: A PCR assay was used to amplify DNA from fecal samples; amplification products were submitted for bidirectional DNA sequencing. Logistic regression was used to test for associations between various host factors and Cryptosporidium spp. RESULTS: 70 of the 115 (61%) fecal samples were found to have C parvum DNA, 42 (37%) were determined to have C bovis DNA, and 3 (3%) were found to have C parvum deer-type DNA. The presence of diarrhea at the time of fecal sample collection, oocyst count, and breed were associated with whether cattle were infected with C parvum or C bovis, with animals more likely to be infected with C parvum if they had diarrhea, had a high oocyst count, or were Holsteins. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that C parvum and C bovis can be isolated from dairy cattle in New York State and that various factors affect whether cattle infected with Cryptosporidium spp are infected with C parvum or C bovis. Findings also lend credence to the theory that C bovis may be more host adapted and thus less pathogenic to dairy cattle than C parvum.     

Effects of colostral antibody on susceptibility of calves to Cryptosporidium parvum infection

Effects of colostral antibody on susceptibility of calves to Cryptosporidium parvum infection were examined. Six calves were fed pooled colostrum that contained C parvum antibody, 6 times daily (at 4-hour intervals) for 7 days and then milk replacer for 7 days. Colostrum was obtained from healthy cows or cows inoculated parenterally with C parvum oocysts before parturition. Antibody content was determined in serum and colostrum whey, using an ELISA for anticryptosporidia immunoglobulin. Six calves were fed colostrum from healthy cows 1 time, and then milk replacer 6 times daily for 14 days. On day 1, all calves were challenge exposed with C parvum, PO, and were monitored daily for diarrhea and oocyst shedding. Bovine colostrum containing specific antibody to C parvum, at ELISA titers up to 10,240, was not effective in protecting calves against challenge exposure to C parvum.     

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.     

Distribution of Cryptosporidium parvum subtypes in calves in Germany

Cryptosporidium DNA was extracted from 134 faecal specimens from pre-weaned calves from different German Federal States (age range, 3–15 days old), which tested positive for oocysts by microscopic analysis. The 18S rDNA gene and the oocyst wall protein gene (COWP) were used as targets for PCR and RFLP techniques. Cryptosporidium species were identified by using SspI, MboII and RsaI endonucleases for the digestion of 18S rDNA and COWP amplified fragments, respectively. In all samples, restriction patterns corresponding to Cryptosporidium parvum were identified, which is in agreement with abundant literature data indicating C. parvum as the most common species in pre-weaned calves. In order to estimate the genetic heterogeneity among C. parvum calf isolates, 53 samples chosen to represent different German Federal States were successfully subtyped by sequence analysis of the highly polymorphic 60-kDa glycoprotein gene. All isolates belonged to the allele IIa (with seven subtypes), with the exception of one isolate that belonged to the allele IId. Moreover, three novel subtypes of the allele family IIa have been found. This study confirms the utility of genotyping and subtyping tools in characterizing the transmission of Cryptosporidium spp. This is the first molecular epidemiological report about subtyping of Cryptosporidium bovine isolates in Germany.     

Malabsorption of vitamin A in preruminating calves infected with Cryptosporidium parvum.

Serum retinol, retinyl palmitate, and total vitamin A concentrations, and jejunoileal morphology were examined in neonatal calves infected with Cryptosporidium parvum. Group-1 calves served as noninfected controls and, after an adjustment period, were given 50 ml of saline solution i.v. every 12 hours for 6 days. Group-2 calves were inoculated with 10(7) C parvum oocysts and, after the onset of diarrhea, were given 50 ml of saline solution i.v. every 12 hours for 6 days. Group-3 calves were inoculated with 10(7) C parvum oocysts and, after the onset of diarrhea, were treated with difluoromethylornithine (DFMO, 200 mg/kg of body weight i.v., q 12 h) for 6 days. Group-4 calves were naturally infected with C parvum. Jejunoileal biopsy specimens were excised from calves of groups 1-3 at 3 and again at 15 to 16 days of age. During the course of diarrhea and 3 days after saline or DFMO administration, water-miscible retinyl palmitate was administered orally (2,750 micrograms/kg) to each calf in each group. Cryptosporidium parvum infection was associated with significant (P < or = 0.05) reduction in postadministration serum retinol, retinyl palmitate, and total vitamin A concentrations in calves of groups 2, 3, and 4. Cryptosporidium parvum infection caused significant (P < or = 0.05) reduction in villus height. Decreased villus height, villus blunting and fusion, and attenuation of the intestinal mucosa were associated with reduced absorption of vitamin A, as indicated by lower peak postadministration retinyl palmitate concentration in C parvum-infected calves. Intravenous administration of DFMO to group-3 calves did not improve retinol absorption.(ABSTRACT TRUNCATED AT 250 WORDS)     

The occurrence of Cryptosporidium parvum,Campylobacter and Salmonella in newborn dairy calves in the Manawatu region of New Zealand

AIM: To determine the occurrence of Cryptosporidium parvum oocysts, Campylobacter spp and Salmonella spp in faecal samples taken from newborn dairy calves on 24 dairy farms in the Manawatu region of New Zealand.

METHODS: A cross-sectional study was conducted during the 2002 calving season. Faecal samples were collected from 185 newborn calves from a convenience sample of 24 dairy farms. The samples were tested microscopically for the presence of C. parvum oocysts, and bacteriologically for the presence of Campylobacter spp and Salmonella spp.

RESULTS: Infections with C. parvum were identified in 33/156 (21.2%) calves from 10 farms. More than 10⁶ oocysts/g (OPG) faeces were detected in calves from four farms. Campylobacter spp were isolated from 58/161 (36%) calves from 18 farms; in particular, C. jejuni subsp jejuni was isolated from 11/161 (6.8%) calves from seven farms. Salmonellae were not detected.

CONCLUSIONS: Despite the short and concentrated calving pattern and the long interval between calving seasons characterising most dairy farms in New Zealand, C. parvum is widespread among calves. Campylobacter spp, especially C. jejuni, rapidly colonise the intestinal tract of newborn calves.

RELEVANCE: This study provided an estimate of the ecological impact of newborn dairy calves with regard to the potentially zoonotic enteric pathogens most frequently isolated from human gastrointestinal infections in New Zealand.     

A rare Cryptosporidium parvum genotype associated with infection of lambs and zoonotic transmission in Italy

An outbreak of cryptosporidiosis occurred in a mixed sheep/cattle farm of Central Italy in October 2011. A total of 450 ovines (250 sheep and 200 lambs) and 140 bovines (130 cows and 10 calves) were housed in two separated units, at the time of the outbreak. About half of the lambs had diarrhea due to Cryptosporidium sp. with a mortality rate of 80%; calves were not infected. Genomic DNA was extracted from an archived slide and from fecal specimens, and the parasite was identified as Cryptosporidium parvum by PCR and sequence analysis at the CpA135 gene. Genotyping at the GP60 gene showed the presence of a very rare genotype, IIaA20G2R1. Shortly after the outbreak was identified, the son of the farm's owner, aged 18 months, experienced an acute gastroenteritis and was hospitalized due to recurrent episodes of diarrhea, fever, vomiting and lack of appetite. The feces tested negative for bacteria and viruses, whereas cryptosporidiosis was diagnosed by microscopy and an immunochromatographic test. Molecular typing identified the C. parvum genotype IIaA20G2R1 in the feces of the child. This is the first case of transmission of cryptosporidiosis in Italy involving lambs as source of oocysts infectious to humans.     

The occurrence of Cryptosporidium parvum, Campylobacter and Salmonella in newborn dairy calves in the Manawatu region of New Zealand

AIM: To determine the occurrence of Cryptosporidium parvum oocysts, Campylobacter spp and Salmonella spp in faecal samples taken from newborn dairy calves on 24 dairy farms in the Manawatu region of New Zealand. METHODS: A cross-sectional study was conducted during the 2002 calving season. Faecal samples were collected from 185 newborn calves from a convenience sample of 24 dairy farms. The samples were tested microscopically for the presence of C. parvum oocysts, and bacteriologically for the presence of Campylobacter spp and Salmonella spp. RESULTS: Infections with C. parvum were identified in 33/156 (21.2%) calves from 10 farms. More than 10(6) oocysts/g (OPG) faeces were detected in calves from four farms. Campylobacter spp were isolated from 58/161 (36%) calves from 18 farms; in particular, C. jejuni subsp jejuni was isolated from 11/161 (6.8%) calves from seven farms. Salmonellae were not detected. CONCLUSIONS: Despite the short and concentrated calving pattern and the long interval between calving seasons characterising most dairy farms in New Zealand, C. parvum is widespread among calves. Campylobacter spp, especially C. jejuni, rapidly colonise the intestinal tract of newborn calves. RELEVANCE: This study provided an estimate of the ecological impact of newborn dairy calves with regard to the potentially zoonotic enteric pathogens most frequently isolated from human gastrointestinal infections in New Zealand.