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。     

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.     

Specific IgA antibody response to coproantigens of Cryptosporidium parvum in serum and saliva of calves after experimental infection

The antibody response to coproantigens of Cryptosporidium parvum was examined in saliva and sera of calves experimentally infected with C. parvum. Coproantigens of C. parvum with approximate molecular masses of 17, 15 and less than 14kDa were found in the feces of infected calves on day 3 or later, and 60 and 23kDa coproantigens observed between days 4 and 9 post-infection, respectively. The antibody reactivity to the coproantigens was mainly attributable to IgA class antibodies in saliva and was detectable during the convalescent phase of infection. A 15kDa protein isolated from the feces of infected calves by immunoaffinity adsorption using a monoclonal anti C. parvum antibody was recognized by IgA antibodies present in the saliva during the convalescent phase of infection. These results suggest that this coproantigen may be released from C. parvum sporozoites and may induce IgA antibody production in the mucosal immune system of infected calves.     

Effect of colostral antibody on bovine leukemia virus infection of neonatal calves

Pairs of newborn calves were exposed to bovine leukemia virus (BLV) when they were given their 1st colostrum feeding. Calves that were given 10(6) BLV-infected lymphocytes in colostrum free of BLV-specific antibody became infected. Calves that were fed 10(7), 10(8), or 10(9) infected lymphocytes in colostrum that contained BLV-specific antibody did not become infected. One of 2 calves inoculated intradermally with 250,000 infected lymphocytes was protected by colostral antibody, but the other was not. Colostral antibody titers in the unprotected calf decreased normally until the calf was 4 months old and then increased markedly; this pattern indicates that the presence of colostral antibody may have prolonged the latent period of the BLV infection.     

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 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.     

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.     

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.     

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.     

Excretion patterns of mucosally delivered antibodies to p23 in Cryptosporidium parvum infected calves

Fecal samples obtained at intervals from six calves with acute cryptosporidiosis contained antibodies of multiple isotypes to p23. IgM-, IgA-, and IgG(1)-isotype anti-p23 appeared before IgG(2)-isotype antibodies. All anti-p23 antibodies had declined by 2 months after infection. One calf that failed to shed oocysts following initial exposure developed IgG(1)-isotype anti-p23 antibodies. One calf that died following exposure to Cryptosporidium parvum oocysts lacked detectable anti-p23 antibodies. Re-inoculation with C. parvum resulted in a brief, marked recall response to p23.     

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.     

Effect of Cryptosporidium parvum infection on the absorptive capacity and paracellular permeability of the small intestine in neonatal calves

Cryptosporidium parvum is recognized as one of the most important pathogens causing enteritis and severe diarrhoea in calves up to 1 month of age. Although the infection may be responsible for some mortality, its impact is mainly associated with the impairment of intestinal functions and lower performance of animals. The aim of this study was to determine the effect of cryptosporidiosis on the intestinal functions in neonatal experimentally infected Holstein calves. Absorption tests with d-xylose and retinyl-palmitate, and the lactulose/mannitol test of intestinal permeability were simultaneously performed in 1-week intervals from challenge to full recovery. In infected animals, reduced intestinal absorptive capacity for both D-xylose and retinyl-palmitate was observed on day 7 post-infection (p.i.). At the same time, a more than 100% elevation of intestinal permeability was observed in the infected calves. All intestinal functions, except absorption of retinyl-palmitate, were significantly affected and changes were detected up to day 14 p.i. In contrast, results of all tests obtained on day 21 p.i. suggest full recovery of the infected intestine. Significantly, growth of the calves which had recovered from cryptosporidiosis was still affected between days 14 and 21 p.i.     

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)     

Cryptosporidium parvum infection in orphan lambs on a farm open to the public

A longitudinal survey was undertaken on an open farm to investigate the occurrence of Cryptosporidium species infection in orphan lambs obtained from three local flocks. During an initial pilot study, Cryptosporidium oocysts were detected by a fluorescent antibody test (fat) in the faeces of two of 21 lambs aged between one and three weeks derived from one flock (flock A). Pooled pen samples of faeces were collected weekly from lambs derived from each flock; oocysts were detected by fat in 24 (49.0 per cent) of 49 samples from lambs from flock A, 18 (30.5 per cent) of 59 samples from lambs from flock B and 14 (29.8 per cent) of 47 samples from lambs from flock C. Oocyst counts of 1 x 10(3) to more than 2 x 10(6) per gram of faeces were detected in lambs up to 12 weeks old, with the peak counts occurring at six weeks of age in the lambs from flocks A and B and at four weeks of age in those from flock C. The oocysts were confirmed by molecular analysis as Cryptosporidium parvum. Virtually all the infections were subclinical.     

Recombinant bovine interleukin-12 stimulates a gut immune response but does not provide resistance to Cryptosporidium parvum infection in neonatal calves

This study was undertaken to determine if administration of recombinant bovine interleukin-12 (rBoIL-12) could stimulate a cellular immune response that protected calves from an oral challenge inoculation with Cryptosporidium parvum oocysts. In a first experiment, rBoIL-12 intraperitoneally administered as a single dose 1 day before challenge inoculation, did not alter the course of infection. The percentage of immune competent cells and levels of cytokine gene expression in the ileo-cecal mucosa and in the draining lymph nodes of treated calves were similar to those of untreated control calves. However, when rBoIL-12 was subcutaneously administered daily from 2 days before infection to 2 days after infection, a consistent increase of T lymphocytes and an higher expression of interferon-gamma (IFN-gamma) was detected. Again, treatment did not alter the course of infection. Similar results were obtained when rBoIL-12 was administered daily for 4 days beginning 2 days after oral inoculation. These data indicate that although rBoIL-12 stimulated a strong immune response in the gut of neonatal calves, the response was not able to provide protection from challenge inoculation with C. parvum oocysts.     

Prevalence of Cryptosporidium infection in calves in France

Two multicentre surveys were conducted in France to estimate the prevalence of Cryptosporidium infection in calves using qualitative ELISA for detection of Cryptosporidium coproantigens and oocysts. The first survey involved 4-12-day-old calves in six dairy-calf distribution centres, collecting calves from seven Administrative Regions (Aquitaine, Bretagne, Franche-Comté, Lorraine, Normandie, Nord, Pays de Loire). For each region, 20 calves were selected every month for 12 consecutive months (October 1995-September 1996). Prevalence of Cryptosporidium infection was 17.9% (Confidence Intervals (C.I.) 95%=[16.1%; 19.8%]) among the 1628 selected calves, of which only 5.3% had diarrhoea. The second survey conducted between November 1995 and May 1996 involved 4-21-day-old calves examined by veterinary practitioners who selected 189 livestock farms of dairy- or suckler-type in ten Administrative Departments (Allier, Cantal, Creuse, Doubs, Ille-et-Vilaine, Maine-et-Loire, Manche, Pas-de-Calais, Sa?ne-et-Loire, Vendée). Cryptosporidia were detected in 105 (55.6%) of the farms. Among the 440 calves examined, of which 398 (90.5%) presented diarrhoea, cryptosporidia were found in 191 animals, i.e. a prevalence of 43.4% (C.I. 95%=[38. 8%; 48.0%]). Breed of calves and type of housing had very little impact on prevalence in this survey. Some regional variations could be noticed, even if cryptosporidia infection is widespread. Monthly variations could be related to seasonal peaks in calving with a lower infection rate during summer.     

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.     

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.     

延边地区牛隐孢子虫感染情况的初步调查

隐孢子虫病是由隐孢子虫 ( Cryptosporidium,CS)寄生于人或动物胃肠道上皮细胞内所引起的一种人兽共患的原虫病。因其在婴幼儿、免疫缺陷患者 ,尤其是艾滋病病人中感染率较高 ,且危害严重而倍受重视。家畜中常见的临床症状为间歇性水泻、脱水和厌食 ,还可能有进行性消瘦和减重。人表现持续性腹泻 ,伴有营养不良、腹痛、发热和呕吐。因此 CS已被确定为引起家畜和人类腹泻的重要病因之一。据国外报道 ,CS在犊牛中感染率相当普遍和严重。由肠道疾病致死的牛中 ,30 %是由 CS造成的 ,因而给畜牧业造成较大的经济损失。在国内 ,于 1 986年陈义…