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1.

Background

Campylobacteriosis is a zoonotic disease, and animals such as poultry, pigs and cattle may act as reservoirs for Campylobacter spp. Cattle shed Campylobacter spp. into the environment and they can act as a reservoir for human infection directly via contact with cattle or their faeces or indirectly by consumption of contaminated food. The aim of this study was to determine the prevalence, the quantitative load and the genetic strain diversity of Campylobacter spp. in dairy cattle of different age groups.

Results

Faecal samples of 200 dairy cattle from three farms in the central part of Lithuania were collected and examined for Campylobacter. Cattle herds of all three farms were Campylobacter spp. positive, with a prevalence ranging from 75% (farm I), 77.5% (farm II) to 83.3% (farm III). Overall, the highest prevalence was detected in calves (86.5%) and heifers (86.2%). In contrast, the lowest Campylobacter prevalence was detectable in dairy cows (60.6%). C. jejuni, C. coli, C. lari and C. fetus subsp. fetus were identified in faecal samples of dairy cattle. C. upsaliensis was not detectable in any sample. The high counts of Campylobacter spp. were observed in faecal material of dairy cattle (average 4.5 log10 cfu/g). The highest numbers of Campylobacter spp. were found in faecal samples from calves (average 5.3 log10 cfu/g), whereas, faecal samples from cows harboured the lowest number of Campylobacter spp. (average 3.7 log10 cfu/g). Genotyping by flaA PCR-RFLP analysis of selected C. jejuni isolates showed that some genotypes were present in all farms and all age groups. However, farm or age specific genotypes were also identified.

Conclusions

Future studies are needed to investigate risk factors related to the degree of colonisation in cattle. Based on that, possible measures to reduce the colonisation and subsequent shedding of Campylobacter in cattle could be established. It is important to further investigate the epidemiology of Campylobacter in the cattle population in order to assess associated risks to public health.  相似文献   

2.
The prevalence of Cryptosporidium spp. infection in a cross-sectional study of dairy cattle, from two contrasting dairying regions in Tanzania, were determined by staining smears of faecal samples with the modified Ziehl-Neelsen technique. Of the 1 126 faecal samples screened, 19.7% were positive for Cryptosporidium spp. The prevalence was lower in Tanga Region than in Iringa Region. The prevalence of affected farms was 20% in Tanga and 21% in Iringa. In both regions, the probability of detecting Cryptosporidium oocysts in faeces varied with animal class, but these were not consistent in both regions. In Tanga Region, Cryptosporidium oocysts were significantly more likely to be found in the faeces of milking cows. In Iringa Region, the likelihood that cattle had Cryptosporidium-positive faeces declined with age, and milking cattle were significantly less likely to have Cryptosporidium-positive faeces. In this region, 7% of cattle were housed within the family house at night, and this was marginally associated with a higher likelihood that animals had Cryptosporidium-positive faeces. Our study suggests that even though herd sizes are small, Cryptosporidium spp. are endemic on many Tanzanian smallholder dairy farms. These protozoa may impact on animal health and production, but also on human health, given the close associations between the cattle and their keepers. Further studies are required to assess these risks in more detail, and understand the epidemiology of Cryptosporidium spp. in this management system.  相似文献   

3.
The prevalence of enterohemorrhagic Escherichia coli (EHEC) O157 was examined in bovine faeces. EHEC O157 was isolated from the faeces of 42 (13.0%) of 324 cattle. Of the 4 farms and the facilities tested, the 3 farms and the facilities were found positive for EHEC O157. The highest isolation rate among the farms was 33.7%. The prevalence of EHEC O157 in heifers was higher than that in calves and other cattle. No cattle positive for EHEC O157 showed any clinical signs except 2 calves with diarrhea in a veterinary hospital. Almost all isolates possessed the stx gene, and Stx-positive strains carrying both stx(1) and stx(2) genes were predominant. These results indicate that EHEC O157 are distributed in bovine faeces, and that dairy and beef farms in selected regions of Japan are heavily contaminated with the organisms.  相似文献   

4.
Faecal samples were collected from 734 cattle selected at random from 60 dairy farms in Galicia (NW Spain). The animals studied were classified into 12 age groups: <1 month (53); 1-5 months (30); 6-11 months (31); 12-16 months (72); 17-20 months (64); 21-24 months (96); 3 years (94); 4 years (74); 5 years (67); 6 years (67); 7-8 years (63) and 9-13 years (23). Oocysts of Cryptosporidium spp. were identified in 104 animals (14.2%) distributed throughout all of the age groups and from 40 different farms (66.7%). The percentage of cattle infected ranged between 58.5% in calves <1 month and 7.9% in 7 to 8-year-old cows, i.e. the percentage of infection decreased significantly (P < 0.05) with increasing age. The intensity of infection in animals older than 1 month ranged between 10 and 5924 oocysts/g of faeces and there were no significant differences between the different groups. Cysts of Giardia duodenalis were identified in 221 animals (30.1%) from 56 farms (93.3%). The parasite was detected in all age groups, at rates of infection ranging between 21.8% (9-13 years) and 56.7% (1-5 months), although these differences were not statistically significant. The intensity of infection ranged between 7 and 15 412 cysts/g of faeces, with the number of cysts shed being significantly higher (P < 0.05) in calves <1 month than in calves aged 1-5 months. Significant associations between parasitisation by Cryptosporidium spp. or G. duodenalis and the consistency of the faeces were only found in calves aged <1 month and 1-5 months. Concurrent infections were more prevalent in the groups of calves of 1-5 months (23.3%) and 6-11 months (25.8%).  相似文献   

5.
In this study, the prevalence and distribution of various Arcobacter spp. were investigated in samples taken from the cloacae of healthy domestic geese raised in Turkey. A membrane filtration technique with a non-selective blood agar was employed after enrichment in Arcobacter enrichment broth (AEB) to isolate a wide range of Arcobacter spp. In addition, the isolates were characterized phenotypically and identified at species level using a multiplex-PCR assay. A total of 90 cloacal swab samples taken from geese, collected on three farms (18, 25, 47 samples, respectively), were examined. Of the samples examined, 16 (18%) were found positive for Arcobacter. One Arcobacter species was isolated from each bird. Of the 16 Arcobacter isolates, 7 (44%), 7 (44%) and 2 (12.5%) were identified by m-PCR as A. cryaerophilus, A. skirrowii and A. butzleri, respectively. The present study indicates that domestic geese can harbour a variety of Arcobacter spp. in their cloacae. The presence of Arcobacter in geese may be of significance as reservoirs in their dissemination. Detailed research is needed for better understanding of the epidemiology and zoonotic potential of this emerging pathogen.  相似文献   

6.
Epidemiologic investigations of Salmonella infections in dairy cattle often rely on testing fecal samples from individual animals or samples from other farm sources to determine herd infection status. The objectives of this project were to evaluate the effect of sampling frequency on Salmonella isolation and to compare Salmonella isolation and serogroup classification among sample sources on 12 US dairy farms sampled weekly for 7-8 weeks. Three herds per state were enrolled from Michigan, Minnesota, New York and Wisconsin based upon predefined herd-size criteria. Weekly samples were obtained from cattle, bulk tank milk, milk filters, water and feed sources and environmental sites. Samples were submitted to a central laboratory for isolation of Salmonella using standard laboratory procedures. The herd average number of cattle fecal samples collected ranged from 26 to 58 per week. Salmonella was isolated from 9.3% of 4049 fecal samples collected from cattle and 12.9% of 811 samples from other sources. Serogroup C1 was found in more than half of the samples and multiple serogroups were identified among isolates from the same samples and farms. The percentage of herd visits with at least one Salmonella isolate from cattle fecal samples increased with overall herd prevalence of fecal shedding. Only the three herds with an average fecal shedding prevalence of more than 15% had over 85% of weekly visits with at least one positive fecal sample. The prevalence of fecal shedding from different groups of cattle varied widely among herds showing that herds with infected cattle may be classified incorrectly if only one age group is tested. Testing environmental sample sources was more efficient for identifying infected premises than using individual cattle fecal samples.  相似文献   

7.
Salmonella in cattle herds may behave as epidemic or endemic infections. An intensive longitudinal sampling study across all management groups and ages on six dairy farms in the UK was used to examine patterns of Salmonella shedding, following the prior identification of either Salmonella Dublin (SD) (three farms) or Salmonella Typhimurium (ST) (three farms) on the premises in the context of clinical salmonellosis. Individual faeces, pooled faeces and environmental samples (total 5711 samples), taken approximately every six weeks for 15-24 weeks, were cultured for Salmonella. SD was detected at low frequency (on any visit, 0.5-18.3 per cent of samples positive) and most consistently in calves. By contrast, ST was isolated at higher frequency (on any visit, 6.8-75 per cent of samples positive), and in higher numbers, up to 10(7) cfu/g faeces. Significantly more samples from calves were positive for ST than were positive for SD (50.6 per cent v 3.1 per cent; P?相似文献   

8.
Coccidial oocysts were detected in 35% of 445 cattle in four medium-scale and 20 small-scale dairy farms in Morogoro municipality, Tanzania. The highest prevalence (56%) was observed in animals aged between 5 and 18 months, whereas lower prevalences were observed in calves (29%) aged between 12 days and 4 months and adults (30%). No coccidial oocysts were detected in calves less than 12 days old. The oocyst output was high in calves, followed by weaners; adults had the lowest oocyst output. The number of oocysts per gram of faeces was significantly higher (P < 0.001) in diarrhoeic animals than in non-diarrhoeic animals, and more so in young calves. Eimeria species infecting the animals included Eimeria bovis (68%) and Eimeria zuemii (57%), Eimeria ellipsoidalis (25%), Eimeria cylindrica (23%), Eimeria auburnensis (22%), Eimeria alabamensis (12%) and Eimeria subspherica (5%). Mixed infections involving two or three species were common. Our findings indicate that eimeriosis is common in cattle in Morogoro municipality.  相似文献   

9.
The prevalence of verocytotoxin-producing Escherichia coli (VTEC) O157 in 12-30-month-old beef finishing cattle in Scotland was determined using 1g faeces samples enriched in buffered peptone water, followed by immunomagnetic separation (IMS) and isolation on sorbitol MacConkey agar with cefixime and tellurite supplement (CT-SMAC). A validated questionnaire was used to collect information that could be associated with the samples. Generalised Linear Models and Generalised Linear Mixed Models were used to identify factors associated with shedding both between and within groups. A total of 14,856 samples were collected from 952 farms, of which 1231 were positive for VTEC O157. Prevalence levels were calculated with 95% confidence intervals as follows: 7.9% (6.5%, 9.6%) of animals sampled were estimated to be shedding VTEC O157, while 22.8% (19.6%, 26.3%) of farms were estimated as having at least one animal shedding in the group sampled. The median percentage of animals shedding in positive groups was 25% (20%, 32%). An increased probability of a group containing a shedding animal was associated with larger numbers of finishing cattle, the presence of pigs on the farm, or the farm being classed as a dairy unit stocking beef animals. Farms that spread slurry on grazing land were more likely to have shedding animals, while those that spread manure were at lower risk. Groups with older animals were less likely to be identified as positive. There was no significant regional difference in group shedding probabilities, but the proportion of positive groups dropped over two successive years of the study. Higher mean levels of shedding in positive groups were associated with animals being housed rather than at pasture, and this effect was stronger in groups which had recently had a change in housing or diet. Farms with animals at pasture had lower mean prevalence where water was supplied from a natural source, as had farms with higher numbers of finishing cattle. There remained unexplained variability in mean prevalence levels on positive farms in different areas of Scotland.  相似文献   

10.
The objective of this study was to develop a multiplex polymerase chain reaction (PCR) to detect and differentiate food-borne pathogens of the three genera Campylobacter, Arcobacter and Helicobacter in a single step procedure. One common reverse primer and three genus-specific forward primers were designed by hybridizing to the 16S rRNA of selected reference strains. Besides the species with significance as food-borne pathogens isolated from poultry meat--Campylobacter jejuni, Campylobacter coli, Arcobacter butzleri and Helicobacter pullorum--several other members of these genera were tested to determine the specificity of the designed multiplex PCR. In total, 20 ATCC and NCTC reference strains of Campyobacter, Arcobacter and Helicobacter were used to evaluate the PCR. Specific amplificates were obtained from all thermophilic species of Campylobacter as well as from species of Arcobacter and Helicobacter. No amplification product was obtained from the non-thermophilic Campylobacter, C. hyointestinalis and C. fetus. Furthermore, a total of 43 field strains of the three genera isolated from poultry, pigs, cattle and humans were investigated using this PCR. To confirm the classification of 10 H. pullorum strains the 16S rRNAs were sequenced. The developed PCR is a helpful diagnostic tool to detect and differentiate Campylobacter, Arcobacter and Helicobacter isolated from poultry and poultry products.  相似文献   

11.
Infections with internal parasites are one of the most important causes of reduced productivity in first-grazing season cattle (FGSC). In conventional herds, nematode infections can be controlled by prophylactic anthelmintic treatments, but this is prohibited in organic production. The purpose of this investigation was to monitor the status of internal parasitism on 15 organic cattle enterprises in southwestern Sweden during the 1997 and 1998 grazing seasons, and to estimate the benefits of some management practices in parasite control. On each farm, the numbers of Eimeria alabamensis oocysts per gram of (opg) faeces were counted in seven fresh dung pats collected from the paddock 8-10 days after the turnout of FGSC. Faecal samples from 5 to 15 FGSC were also analysed for nematode eggs per gram (epg) faeces at four occasions during each grazing season. In addition, the FGSC and one group of second-grazing season cattle (SGSC) were weighed at turnout and housing and at the same time blood samples were collected, and analysed for serum pepsinogen concentration and antibodies against the lungworm, Dictyocaulus viviparus. On seven farms, 1-6 samples with more than 100000 opg were found, indicating considerable pasture contamination by E. alabamensis. However, clinical signs of coccidiosis were not observed. The highest outputs of nematode eggs were observed 45-55 days after turnout. More than 500 epg were only observed in 12 (2.2%) of the calves in 1997 and in three (0.6%) animals in 1998. Only 1% of the serum samples had pepsinogen values exceeding 3.6 U tyrosine, indicative of subclinical Ostertagia ostertagi infection. Lungworm infection was detected in five and nine herds in 1997 and 1998, respectively. The number of seropositive animals on these farms ranged between one (10%) and seven (70%). Clinical signs of dictyocaulosis were observed on two farms. The results indicate that dictyocaulosis is a problem in organic dairy herds in Sweden. On the other hand, the study shows that good management such as usage of parasite safe pastures and supplementary feeding may help control gastrointestinal parasites.  相似文献   

12.
The present study was carried out to assess the prevalence of motile Aeromonas spp. in the faeces of clinically healthy sheep, cattle and horses and evaluate their susceptibility to some anti-microbial agents. Rectal swabs from 120 sheep, 85 cattle and 20 horses were examined for Aeromonas species using alkaline peptone water (pH 8.4) as the enrichment medium and Aeromonas Selective Agar containing 5 mg/l ampicillin as the isolation medium. Identification and antibiotic resistance of motile Aeromonas strains was performed using Gram Negative Enteric ID panel. Motile aeromonads were isolated from 12 (10%) sheep, 7 (8.2%) cattle and 1 (5%) horse. Of these 20 aeromonad isolates, 13 were A. caviae, 6 were A.sobria and 1 was A. hydrophila. Aeromonas species in the faeces of livestock might pose a public health problem for humans who are in direct contact with contaminated animals. However, further studies should be performed on aeromonads relating to their transmission between animals and humans.  相似文献   

13.
Cattle are a natural reservoir for Shiga toxigenic Escherichia coli (STEC), however, no data are available on the prevalence and their possible association with organic or conventional farming practices. We have therefore studied the prevalence of STEC and specifically O157:H7 in Swiss dairy cattle by collecting faeces from approximately 500 cows from 60 farms with organic production (OP) and 60 farms with integrated (conventional) production (IP). IP farms were matched to OP farms and were comparable in terms of community, agricultural zone, and number of cows per farm. E. coli were grown overnight in an enrichment medium, followed by DNA isolation and PCR analysis using specific TaqMan assays. STEC were detected in all farms and O157:H7 were present in 25% of OP farms and 17% of IP farms. STEC were detected in 58% and O157:H7 were evidenced in 4.6% of individual faeces. Multivariate statistical analyses of over 250 parameters revealed several risk-factors for the presence of STEC and O157:H7. Risk-factors were mainly related to the potential of cross-contamination of feeds and cross-infection of cows, and age of the animals. In general, no significant differences between the two farm types concerning prevalence or risk for carrying STEC or O157:H7 were observed. Because the incidence of human disease caused by STEC in Switzerland is low, the risk that people to get infected appears to be small despite a relatively high prevalence in cattle. Nevertheless, control and prevention practices are indicated to avoid contamination of animal products.  相似文献   

14.
A survey was conducted to examine the distribution of Arcobacter species among livestock in Japan. During May 1999 and May 2000, fecal samples from cattle (n=332) and swine (n=250), chicken cloacal swabs (n=234), and vaginal swabs of cattle (n=61) and swine (n=15) were submitted for the isolation of Arcobacter species. Arcobacter species were isolated from 3.6 and 10.0% of the cattle and swine fecal samples, respectively, along with 14.5% of chicken cloacal swabs. No significant seasonal differences were observed. Species-specific polymerase chain reaction assay showed that A. butzleri was the most prevalent species (83.3, 60.0 and 47.1% of the cattle, swine and chicken isolates, respectively), followed by A. cryaerophilus 1B (16.7, 36.0 and 55.9% of the cattle, swine and chicken isolates, respectively). Of the samples from vaginal swabs, 8.1 and 13.3% were positive for Arcobacter in cattle and swine, respectively. This is the first report demonstrating the distribution of Arcobacter species among livestock in Japan.  相似文献   

15.
The prevalence of Blastocystis sp. was examined in fecal samples collected from cattle, pigs, dogs, and a variety of zoo animals (primates, carnivores, herbivores, pheasants, and ducks) by direct observation of fresh fecal suspensions or cultured materials, using light microscopy. The cattle and pigs were randomly sampled from 11 and 12 commercial farms, respectively, located in the western region of Japan. The dog material used in this study was obtained from pets housed in an animal shelter in the city of Osaka. Zoo animals were chosen based on housing conditions that minimized the possibility of intra-zoo transmission of the organism. The prevalence rate among the groups varied greatly. A high prevalence of infection was observed in the farm animal group, ranging from 95% (58/61) in pigs to 71% (39/55) in cattle, whereas the dog fecal samples were completely free of the organism. Prevalence of the organism in the zoo animal were 85% (29/34) in primates, 80% (8/10) in pheasants, 56% (9/16) in ducks, and 0% (0/58) in various carnivores and herbivores. Among the zoo animals infected with Blastocystis, eight species of primates, eight species of pheasants, and four species of ducks were confirmed as new hosts. Since Blastocystis organisms isolated from various animals were morphologically indistinguishable from Blastocystis hominis by light microscopy, further genomic studies are required for analysis of the zoonotic potential or etiological significance of these isolates.  相似文献   

16.
Giardia duodenalis and Cryptosporidium spp. are intestinal protozoan parasites that infect a wide range of host species, including humans. Molecular characterization of these parasites has demonstrated that a number of genotypes and species are common to both humans and animals, and that zoonotic transmission may occur. Numerous studies have reported a high prevalence of G. duodenalis and Cryptosporidium spp. in cattle, particularly calves, and these animals are frequently associated with zoonotic transmission. In the present study, a total of 143 faecal samples from adults, heifers and calves were collected from two dairy cattle farms in eastern Ontario, Canada. The prevalence and molecular characteristics of G. duodenalis and Cryptosporidium spp. in these animals were determined in order to investigate the potential for transmission between cattle and humans in this region. Following DNA extractions from faecal samples, nested-PCR protocols were used to amplify fragments of the 16S rRNA gene and the heat-shock protein 70 (HSP-70) gene for determining the prevalence of G. duodenalis and Cryptosporidium spp. infections, respectively. Genotypes of G. duodenalis, and species of Cryptosporidium, were determined by means of DNA sequencing of amplicons, and subsequent sequence alignment. Cattle on both farms showed a high prevalence of G. duodenalis (42.0%) and Cryptosporidium spp. (27.3%). G. duodenalis infections were more prevalent in calves and heifers than in adults, and Cryptosporidium spp. infections were only observed in calves and heifers. The zoonotic genotype, G. duodenalis Assemblage B was isolated from 24.5% of the cattle tested, while G. duodenalis Assemblage E was found in 17.5% of the cattle tested. The overall prevalence of the zoonotic species Cryptosporidium parvum in the animals tested was found to be 21.7%, while only 1.4% were infected with C. bovis. These findings suggest that there is a potential risk of zoonotic and/or zooanthroponotic transmission of G. duodenalis and C. parvum infections between cattle and humans in eastern Ontario, likely by means of contaminated water or food, or through direct faecal-oral transmission in the case of farmers and veterinary staff.  相似文献   

17.
Eleven cattle farms, 8 layer farms, 7 broiler farms and 30 broiler meat samples were investigated in south-eastern Italy throughout 2003 to evaluate the prevalence, the molecular type and antimicrobial resistance of thermophilic Campylobacters. A total of 398 samples were analysed. One Campylobacter isolate for each positive faecal swab and three isolates per positive broiler meat sample were selected for further analysis. Multiplex PCR was performed for species-level identification and PCR-RFLP of the flagellin A gene for genotyping. Resistance to 14 antimicrobials was studied in 188 Campylobacter isolates. Prevalence of campylobacters was high both on farms (100%) and in food samples (73%). On 4/11 cattle farms and on 10/15 poultry farms more than one species was isolated. The presence of more than one genotype was found on 8/11 cattle farms, on 10/15 poultry farms and in 8/22 Campylobacter-positive food samples. High rates of resistance to quinolone were observed: 9/31 (29%) C. jejuni bovine isolates, 4/22 (18%) C. jejuni poultry isolates, and 14/26 (54%) C. coli poultry isolates. Resistance to sulphamethoxazole-trimethoprim was also observed frequently: 18/26 (69%) of the avian C. coli strains, 25/31 (80%) of the C. jejuni strains isolated from poultry and 15/22 (68%) of those isolated from cattle were resistant. There was a significant difference between the rate of resistance to macrolides of C. coli and C. jejuni isolated in poultry, which amounted to 23% and 3%, respectively. This study provided data on the prevalence and antimicrobial resistance of thermophilic campylobacters in south-eastern Italy and confirmed that flaA-typing is an efficient tool to study the epidemiology of Campylobacter strains in short-term investigations.  相似文献   

18.
The objective of this study was to compare the occurrence and the genotypes and species of Giardia duodenalis and Cryptosporidium spp. in beef and dairy cattle from farms in the Regional Municipality of Waterloo, Ontario, in an effort to determine the potential for zoonotic transmission from these animals. Pooled manure samples were collected from 45 dairy cattle farms and 30 beef cattle farms. The presence of Giardia cysts and Cryptosporidium oocysts was determined by immunofluorescence microscopy, while nested-PCR and DNA sequencing were used to determine genotypes and species. The overall farm prevalence was very high for both Giardia and Cryptosporidium, and was similar for dairy cattle farms (96 and 64%, respectively) and beef cattle farms (97 and 63%, respectively). However, on dairy cattle farms, G. duodenalis and Cryptosporidium spp. were detected in 44% and 6% of total pooled pen manure samples, respectively, with the occurrence of both parasites being generally higher in calves than in older animals. Most Giardia isolates were identified as either the host-adapted genotype G. duodenalis Assemblage E or the zoonotic Assemblage B. Cryptosporidium parvum and Cryptosporidium andersoni were the most frequently identified species in dairy cattle, while the non-zoonotic species Cryptosporidium ryanae and Cryptosporidium bovis were also found. On beef cattle farms, 72% and 27% of the total pooled pen manure samples were positive for Giardia and Cryptosporidium, respectively, with no obvious correlation with age. All Giardia isolates in beef cattle were identified as G. duodenalis Assemblage E, while all Cryptosporidium isolates were identified by sequence analysis as C. andersoni, although microscopic analyses, and subsequent restriction fragment length polymorphism analyses, indicated that other Cryptosporidium species were also present. The results of this study indicate that although Giardia and Cryptosporidium were identified in a higher overall percentage of the pooled beef cattle manure samples than in dairy cattle, firmly established zoonotic genotypes and species were much more common in dairy cattle than in beef cattle in this region. Dairy cattle, and especially dairy calves, may, therefore, pose a greater risk of infection to humans than beef cattle. However, these results may also provide evidence of potential zooanthroponotic transmission (human to animal).  相似文献   

19.
OBJECTIVE: To identify the optimum pooling rate for pooled faecal culture (PFC) as a diagnostic tool in bovine Johne's disease control, for detection of cattle shedding low concentrations of Mycobacterium avium subsp paratuberculosis (Map). METHOD: Thirteen target animals were selected by delayed growth of Map from initial individual radiometric faecal cultures (first growth index at 5 weeks or later). A procedure based on radiometric culture and IS900 polymerase chain reaction and restriction endonuclease analysis confirmation was then used for PFC. RESULTS: Eight samples (stored for up to 17 months at -80 degrees C) yielded Map on subsequent culture, either from undiluted faeces or those mixed with normal cattle faeces at dilution rates from 1 in 5 to 1 in 50. From a regression equation, culture-positive animals were considered to be shedding relatively low levels of Map (< 6 x 10(4)/g of faeces). Pooling dilutions of more than 1 in 5 reduced PFC sensitivity. A minimum incubation period of 10 weeks at a dilution of 1 in 5 is recommended to detect such infected cattle. This pooling rate in radiometric culture is probably capable of detecting cattle shedding < or = 5 x 10(3) Map organisms/g of faeces, representing an estimated inoculum per culture vial of fewer than 20 viable organisms. CONCLUSION: Map was detected in more than 50% of the stored faecal samples from cattle shedding low concentrations of the organism. A pooling rate of 5 samples per pool is required to reliably detect infected low-shedder cattle using PFC based on radiometric culture.  相似文献   

20.
To determine the prevalence of Giardia genotypes in 12–24 month old dairy heifers, fecal specimens were collected from two farms each in Vermont, New York, Pennsylvania, Maryland, Virginia, North Carolina, and Florida. Specimens, cleaned of fecal debris and concentrated using CsCl density gradient centrifugation, were subjected to PCR and DNA sequence analysis. Prevalence of Giardia infection, ranged from 11% to 75% on 14 farms with an average prevalence of 36% (204 positive cattle out of 571 examined). DNA sequence analysis of the 16S rRNA gene revealed 91% of the 204 Giardia isolates were Assemblage E, and 9% were Assemblage A. The prevalence of these genotypes varied greatly from farm to farm, with four farms having exclusively Assemblage E Giardia. Overall, Assemblage E was present in 33% of all animals tested and Assemblage A was present in 3% of the animals. Thus, while many of the heifers were infected with a genotype that is not known to be infectious for humans, 1–2 year old heifers on 10 of 14 farms did harbor zoonotic Assemblage A Giardia. Therefore, heifers cannot be overlooked as a potential source of human infectious cysts in the environment, with some farms representing a much higher risk than others.  相似文献   

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