Natural infection with zoonotic subtype of Cryptosporidium parvum in Capybara (Hydrochoerus hydrochaeris) from Brazil

A total of 145 capybara (Hydrochoerus hydrochaeris) fecal samples from the state of S?o Paulo, Brazil, were screened for Cryptosporidium spp. oocysts using the malachite green method. Eight samples (5.52%) showed positive results and were further submitted to nested PCR reaction for amplification of fragments of 18S rRNA gene and 60-kDa glycoprotein gene for determination of species, alleles and subtypes of Cryptosporidium. Sequencing of the PCR products of the 18S rRNA gene fragments and 60-kDa glycoprotein gene fragments showed that for both genes all Cryptosporidium isolates from capybara were respectively 100% genetically similar to a bovine isolate of C. parvum and to C. parvum subtype IIaA15G2R1. To the best of our knowledge this is the first report of Cryptosporidium infection in this rodent. The finding of zoonotic C. parvum infection in a semi-aquatic mammal that inhabits anthroponotic habitats raises the concern that human water supplies may be contaminated with zoonotic Cryptosporidium oocysts from wildlife.     

Levels of detection of Cryptosporidium oocysts in mussels (Mytilus galloprovincialis) by IFA and PCR methods

Cryptosporidium spp. are monoxenous protozoan parasites that cause gastrointestinal diseases in humans and animals. Shellfish harvesting areas can become contaminated by the infectious stage of the parasite and humans are therefore at risk of infection either by consumption of shellfish, or by taking part in recreational activities in these areas. In the present study we determined the levels of detection, by IFA and PCR techniques, of Cryptosporidium oocysts in mussels experimentally contaminated with a theoretical number of oocysts. There was a significant correlation between the results obtained by both techniques (P<0.05). IFA and PCR were also applied to a total of 222 samples of mussels (Mytilus galloprovincialis) destined for human consumption. In the naturally contaminated samples, we detected a 31.1% of contamination and only Cryptosporidium parvum (previously denominated C. parvum genotype II) was identified.     

A questionnaire-based evaluation of the veterinary cordon fence separating wildlife and livestock along the boundary of the Kruger National Park, South Africa

Veterinary cordon fences are used in Southern Africa to separate wildlife from domestic animals in order to prevent transmission of infectious diseases. Such fences are a control method recognized by the World Organization for Animal Health (OIE) for establishing disease-free zones in beef exporting countries. However, few studies have evaluated the ecological impact of these physical barriers or their effectiveness at a multispecies level. We examined the permeability of one such barrier, along 357 km of the western and southern boundary of the Kruger National Park (KNP) during 2007. Information was gathered using a semi-structured questionnaire implemented among 32 teams of fence maintenance workers. Data were analyzed to identify (a) the main causes of fence damage, (b) the seasonality, location and duration of fence repairs, (c) high permeability areas for elephant (Loxodonta africana), kudu (Tragelaphus strepsiceros), impala (Aepyceros melampus), buffalo (Syncerus caffer) and warthog (Phacochoerus africanus), and (d) the influence of fence electrification, rivers and elephant damage on the frequency of observation of wildlife species outside the KNP estimated during the year. Human and elephant damage were the most common reasons for fence repairs. Elephant and buffalo were the most and least common large mammal species reported observed outside the KNP (1076 and 162 reports/year), respectively. Cattle incursions into the KNP were also reported in 44% of the fence sections. Electrification of the fence was an important factor explaining differences in estimated wildlife species observations outside the KNP during the year. Correlations between estimations of observed species suggested that fence gaps created by elephants might be used by the other wildlife species. Estimated annual counts of kudu, impala and buffalo, but not warthog, were found to correlate with elephant observations. Negative binomial regression models were developed to explore the relationships between observed estimations of different wildlife species outside the KNP the fence, electrification of fence sections and the presence of watercourses, suggesting that kudu, impala and buffalo could use elephant induced fence damage to leave the KNP. The questionnaire was able to evaluate fence integrity and identify sections where integrity was sub-optimal for separating wildlife from domestic livestock species. If combined with more quantitative methods and applied on a routine basis, it could provide an efficient and cost-effective method for monitoring the effectiveness of physical barriers to contain wildlife within protected areas such as is the case with veterinary cordon fences in Southern Africa.     

Fecal shedding of Cryptosporidium oocysts in healthy alpaca crias and their dams

Objective-To determine the apparent prevalence of shedding of Cryptosporidium spp in healthy alpaca crias and their dams on 14 farms in New York and 1 farm in Pennsylvania. Design-Cross-sectional study. Animals-110 alpaca crias and their 110 dams. Procedures-Fecal samples were obtained from 220 alpacas at 14 alpaca farms in New York and 1 farm in Pennsylvania. For each animal, age, sex, and health condition were recorded. A fecal score (1 = normally formed; 2 = soft or loose; 3 = diarrhetic) was recorded for each cria. Cryptosporidium oocysts were identified in fecal samples by a direct immunofluorescence assay. Results-Apparent prevalence of fecal shedding of Cryptosporidium oocysts was 8% (95% confidence interval, 4% to 15%) in dams and was 7% (95% confidence interval, 3% to 13%) in crias. There was no significant difference in age between dams with positive fecal test results for Cryptosporidium oocysts (median age, 4 years; range, 3 to 8 years) and dams with negative results (median age, 4 years; range, 2.5 to 19 years). No significant difference was found in age between crias with positive fecal test results (median age, 20 days; range, 7 to 53 days) and those with negative results (median, 36 days; range, 2 to 111 days). No significant difference in fecal scores was found between crias with positive versus negative fecal test results. Conclusions and Clinical Relevance-A higher than previously reported apparent prevalence of fecal shedding of Cryptosporidium oocysts in healthy alpacas was found. A zoonotic risk should be considered, especially for Cryptosporidium parvum.     

Prevalence of selected zoonotic and vector-borne agents in dogs and cats in Costa Rica

To estimate the prevalence of enteric parasites and selected vector-borne agents of dogs and cats in San Isidro de El General, Costa Rica, fecal and serum samples were collected from animals voluntarily undergoing sterilization. Each fecal sample was examined for parasites by microscopic examination after fecal flotation and for Giardia and Cryptosporidium using an immunofluorescence assay (IFA). Giardia and Cryptosporidium IFA positive samples were genotyped after PCR amplification of specific DNA if possible. The seroprevalence rates for the vector-borne agents (Dirofilaria immitis, Borrelia burgdorferi, Ehrlichia canis, and Anaplasma phagocytophilum) were estimated based on results from a commercially available ELISA. Enteric parasites were detected in samples from 75% of the dogs; Ancylostoma caninum, Trichuris vulpis, Giardia, and Toxocara canis were detected. Of the cats, 67.5% harbored Giardia spp., Cryptosporidium spp., Ancylostoma tubaeforme, or Toxocara cati. Both Cryptosporidium spp. isolates that could be sequenced were Cryptosporidium parvum (one dog isolate and one cat isolate). Of the Giardia spp. isolates that were successfully sequenced, the 2 cat isolates were assemblage A and the 2 dog isolates were assemblage D. D. immitis antigen and E. canis antibodies were identified in 2.3% and 3.5% of the serum samples, respectively. The prevalence of enteric zoonotic parasites in San Isidro de El General in Costa Rica is high in companion animals and this information should be used to mitigate public health risks.     

Age-related and housing-dependence of Cryptosporidium infection of calves from dairy and beef herds in South Bohemia, Czech Republic

Data of the prevalence, age-related and housing-dependence of naturally acquired cryptosporidiosis on 11 dairy and 11 beef farms in South Bohemia (Czech Republic) were collected. The farms were visited over four consecutive years (from 2002 to 2005). The prevalence of Cryptosporidium in pre-weaned (animals until second month of age) and post-weaned (animals from the third month of age) calves was determined. A total of 7001 faecal samples were collected, concentrated by Sheather's floatation method and stained by aniline-carbol-methyl violet. All samples were examined by light microscopy. Cryptosporidium parvum and C. andersoni oocysts were differentiated on morphological criteria. Of the 7021 specimens, 1814 (25.8%) were positive for Cryptosporidium oocysts; 561 samples (8%) for C. parvum and 1253 (17.8%) for C. andersoni. Pre-weaned dairy calves had higher infection levels of C. parvum than pre-weaned beef calves. The prevalence of C. parvum ranged from 1.4 to 56.5% on dairy farms. Only three cases of C. parvum oocysts shedding in pre-weaned calves on beef farms were found. Only one case of C. andersoni infection in pre-weaned calves was detected and no infections of C. parvum in post-weaned calves were found. The prevalence of C. andersoni reached 35.5% on dairy farms and 61.7% on beef farms. Calves that were on pasture all year long, had a lower probability of C. andersoni infection than those calves kept in a cowshed during the winter season.     

Pathogenic bacteria and parasites in wildlife and surface water

In the period October 2003 to August 2005, 897 faecal samples were collected from wild animals and examined for Salmonella spp., Campylobacter spp., and Shiga toxin producing Escherichia coli (STEC) O157, the prevalence of which was found to be 0.1%, 13.8%, and 0.5 %, respectively. Campylobacter spp. were isolated mainly from faecal samples collected from corvidae (59.8%), and meadow birds and waterfowl (22.4%). A subset of these samples was also examined for Cryptosporidium and Giardia oocysts and cysts. None of the 247 samples examined contained C. parvum oocysts, and only 1 sample (roe faeces) contained G. lamblia assemblage A cysts. In the period September to November 2006, samples of running or still surface water were collected at 10 sites on 5 days, to investigate the presence of Salmonella spp., Campylobacter spp., and STEC O157. Twenty (40.8%) of the surface water samples were positive for one or more bacterial pathogens. Seven (14.3%) samples were positiveforSalmonella spp., 14 (28.6%) samples were positive for Campylobacter spp., and 1 (2.0%) sample was positivefor E. coli O157. Samples collected at only 2 of the 10 sites were negative for the pathogens tested; samples collected at the other 8 sites were positive for the pathogens at least once. To gain a better picture of the potential human health risk, this study should be followed up with a more quantitative study of the occurrence of human pathogens in wildlife, taking into account the different natural habitats and behaviour of the different animal populations and a possible seasonal effect. Furthermore, the contamination of surface water with human pathogens should be investigated more extensively.     

Peri-parturient rise of Cryptosporidium oocysts in cows: New insights provided by duplex quantitative real-time PCR

In order to clarify if a peri-parturient rise of Cryptosporidium parvum oocysts occurs in cows, faecal samples from 42 cows on two farms were collected. These samples were taken during the pre-parturient, the peri-parturient and the post-parturient periods. Two methods were used to detect the oocysts, a nested-PCR coupled with sequencing and a duplex real-time PCR (qPCR) that quantified Cryptosporidium spp. DNA concentration. The qPCR results were adjusted using a hierarchical Bayesian model taking into account within and between run variation. Generalised Estimating Equation models (GEE) were used to determine if peri-parturient cows were at greater risk of being infected than pre- or post-parturient cows. Fourteen dairy cows exhibited a peri-parturient and post-parturient rise in the excretion of Cryptosporidium spp. oocysts, other than the zoonotic C. parvum. The cows in the suckler beef farm were the only ones infected with the zoonotic species C. parvum at calving. Due to the low concentration of oocysts excreted mainly from species other than C. parvum, it would appear unlikely that cows act as a source of infection for their calves or contribute significantly to environmental contamination.     

Cryptosporidium parvum in diarrheic calves detected by microscopy and identified by immunochromatographic and molecular methods

Cryptosporidium is an important protozoan parasite that causes diarrhea in neonates and young bovines. The objective of the present study was to determine the frequency of Cryptosporidium infection in animals of dairy farms of the Metropolitan Region (Santiago), Chile. Fecal samples of 205 newborn calves with diarrhea were studied and used for comparing the efficiency of two microscopic staining methods for diagnosis of the parasite, the auramine (AU) and a modified Ziehl-Neelsen (ZN) procedure. Out of the 205 fecal samples, we detected oocysts in 115 (56.1%) with AU and 102 (49.8%) with ZN. Comparison of results obtained with the two microscopic techniques showed significant difference (p<0.05), AU being more sensitive. On the other hand, concordance between the two methods was almost perfect (kappa value of 0.83). The results with these two operator dependent methods were confirmed using an operator independent immunochromatographic (IC) method. The IC method also enabled us to determine the identity of the parasite species as that of Cryptosporidium parvum. Identification of the parasite species was further corroborated by performing a Cryptosporidium species-specific polymerase chain reaction (PCR) test on few samples taken at random. Overall, the results showed a high number of infected animals suggesting the parasite C. parvum as a major parasitic disease agent of neonatal calves with diarrhea in dairy farms of the Metropolitan Region (Santiago) of Chile.     

合肥野生动物园灵长类动物隐孢子虫感染情况调查

[目的]调查合肥野生动物园灵长类动物隐孢子虫的感染情况。[方法]采用饱和蔗糖水漂浮法对环尾狐猴、赤猴和狒狒等6种灵长类动物粪便进行卵囊浓集,采用抗酸染色法对其卵囊染色后进行形态学观察。在形态学观察基础上,对上述疑似粪样中的卵囊进行DNA提取,并采用PCR技术扩增隐孢子虫卵囊壁蛋白基因(COWP),以1%琼脂糖凝胶电泳鉴定PCR扩增产物,并对该园内灵长类动物感染隐孢子虫情况作统计分析。[结果]经形态学鉴定,初步判定从该动物园灵长类动物粪便中获得的卵囊大小为4.66μm×5.18μm,与报道的隐孢子虫形态特征相一致;利用PCR技术扩增得到的目的基因大小为377bp,与预期结果相一致。数据统计表明:合肥野生动物园灵长类动物隐孢子虫感染率为16.67%。[结论]合肥野生动物园灵长类动物存在隐孢子虫感染情况,具有感染人畜的潜在风险。     

Comparison of fecal samples collected per rectum and off the ground for estimation of environmental contamination attributable to beef cattle

OBJECTIVES: To determine whether sampling feces off the ground replicates prevalence estimates for specific pathogens obtained from fecal samples collected per rectum of adult cows, and to determine characteristics of feces on the ground (fecal pats) that are associated with subsequent identification of Campylobacter spp, Cryptosporidium parvum, and Giardia duodenalis. ANIMALS: A random sample of adult beef cattle from 25 herds located throughout California. PROCEDURE: 1,115 rectal and ground fecal samples were obtained. Samples were submitted for culture of Campylobacter spp and examined, using a direct fluorescent antibody assay, to detect C parvum oocysts and G duodenalis cysts. Characteristics of fecal pats, such as volume and consistency, were recorded. RESULTS: Prevalence of Campylobacter spp was 5.0% (20/401) for rectal fecal samples, which was significantly greater than prevalence determined for ground fecal samples (2/402; 0.5%). Most isolates were C jejuni subsp jejuni. Prevalence of C parvum was higher in rectal fecal samples (6/557; 1.1%) than in ground fecal samples (1/558; 0.2%), but this difference was not significant. Prevalence of G duodenalis did not differ for rectal (36/557; 6.5%) versus ground (26/558; 4.7%) fecal samples. CONCLUSIONS AND CLINICAL RELEVANCE: Evaluation of ground fecal samples may not accurately indicate the prevalence of Campylobacter spp or C parvum in cattle but may reflect prevalence of G duodenalis. Differences in prevalence estimates between the 2 methods suggest inactivation of pathogens in feces after cattle have defecated. Prevalence estimates generated by evaluation of ground fecal samples, however, may more accurately estimate environmental pathogen burden.     

Cryptosporidium infection in non-human hosts in Malawi

Of 1346 faecal samples from the Chikwawa and Thyolo districts of Malawi, analysed for the presence of Cryptosporidium oocysts between October 2001 and May 2003, 61.3% were from cattle (29.8% of these were from calves <6 months old). Cryptosporidium oocysts were detected during all three seasons studied in Chikwawa and Thyolo. In Chikwawa, 13.6% of adult cattle and 11.7% of calves were infected, compared to 28.9% of adult cattle and 36.7% of calves in Thyolo. Dependent on season, between 7.8% and 37.7% (Chikwawa) and 16.7% and 39.3% (Thyolo) of cattle samples contained oocysts. In Chikwawa, the highest percentage of infections occurred in the cool season, whereas in Thyolo, the highest percentage of infections occurred in the dry season. Faecal samples from goats [n=225], pigs [n=92], sheep [n=6]), rabbits, guinea pigs, chickens, ducks, turkeys, doves and guinea fowls were also analysed. Up to 5.6% of goat samples contained oocysts in Chikwawa, compared to between 16.7% and 39.3% in Thyolo. Again, in Chikwawa, the highest percentage of infections occurred in the cool season and the lowest in the rainy season, whereas, in Thyolo, the highest percentage of infections occurred in the dry season and the lowest in the cool season. In pigs, more infections were detected in the dry season in Chikwawa, but infections in the cool season were similar (17.7%), whereas in Thyolo, infections occurred in all three seasons (17.9% in the rainy season, 25% in the cool season and 60% in the dry season). Of ten diarrhoeic, oocyst positive cattle faecal samples collected from Chikwawa and subjected to PCR-RFLP, four oocyst positive samples (two from heifers, one from a cow and one unknown) were amplified at an 18S rRNA and Cryptosporidium oocyst wall protein (COWP) loci. RFLP of the 18S rRNA locus indicated that Cryptosporidium parvum, Cryptosporidium hominis, Cryptosporidium bovis and/or Cryptosporidium ryanae DNA, or a mixture of them was present. Cryptosporidium parvum DNA was identified in one sample that amplified at the COWP locus, indicating the presence of the major zoonotic Cryptosporidium species in Malawi.     

Giardia and Cryptosporidium in dairy calves in British Columbia.

A study was undertaken to determine the prevalence of Giardia infections in dairy calves and to compare Giardia and Cryptosporidium infections in calves of different ages. Fresh fecal samples were collected from 386 male and female Holstein calves (newborn to 24 wk) in 20 dairies located in the lower Fraser river valley area of British Columbia. Giardia intestinalis, Cryptosporidium parvum, and Cryptosporidium muris were enumerated in each sample after concentration by sucrose gradient centrifugation and immunofluorescent staining. Giardia was identified at all farm locations. The overall prevalence of Giardia in calves was 73% with a geometric mean cyst count of 1180 cysts per gram of feces (CI, 41 to 5014). Cryptosporidium parvum and C. muris were identified in 80% and 40% of the farms, respectively. The prevalence of C. parvum was 59%, and the geometric mean for oocysts was 457 oocysts per gram of feces (CI, 18 to 160). The prevalence of C. muris was only 2% and the mean oocyst counts were 54 oocysts per gram of feces. Giardiasis was not age dependent, and approximately 80% of the calves from 2 to 24 wk were infected. In contrast, C. parvum infections were predominant in calves 2 to 4 wk, while C. muris was demonstrated in calves older than 4 wk. Fourty-seven percent of calves with diarrhea had high numbers of Giardia cysts in their feces. Giardia infections are highly prevalent in dairy calves and should be considered in animals with diarrhea or failure to thrive.     

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.     

Evaluation of two rapid assays for detecting Cryptosporidium parvum in calf feces

OBJECTIVE: To evaluate 2 rapid, patient-side assays for detection of Cryptosporidium parvum in feces from neonatal calves with diarrhea. DESIGN: Diagnostic test evaluation Sample Population-Fecal samples from 96 neonatal (1 to 30 days old) calves with diarrhea. PROCEDURE: Results of the rapid assays were compared with results of microscopic examination of fecal smears that had been stained with diamant fuchsin stain. RESULTS: One of the rapid assays correctly identified 56 of 62 (90%) fecal samples positive for C. parvum oocysts and 33 of 34 (97%) fecal samples negative for oocysts. The other assay correctly identified 53 of 62 (85%) fecal samples positive for oocysts and 33 of 34 (97%) fecal samples negative for oocysts. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that these 2 rapid assays are accurate when used to detect C. parvum in fecal samples from neonatal calves with diarrhea.     

Presence and molecular characterisation of Giardia and Cryptosporidium in alpacas (Vicugna pacos) from Peru

The presence of Giardia and Cryptosporidium was investigated in 274 faecal samples of alpacas (Vicugna pacos) from 12 herds from Peru by immunofluorescence microscopy and PCR amplification and sequencing of fragments of the ssu-rRNA and β-giardin genes from Giardia spp., as well as the ssu-rRNA gene from Cryptosporidium spp. A total of 137 samples (50.0%) were positive for Giardia spp., and 12 samples (4.4%) for Cryptosporidium spp. In ten samples (3.6%), co-infection by both pathogens was found. Herd prevalence was found to be 91.7% (11/12 herds) for Giardia and 58.3% (7/12 herds) for Cryptosporidium. Regarding the age of the animals, although Giardia was detected in animals as young as 1 week, the prevalence increased with age, reaching 80% by 8 weeks. Similarly, the highest percentage of Cryptosporidium detection (20%) was also found in the 8 week-old group. By PCR, 92 of the 274 analysed samples were positive for Giardia. Sequencing of the amplicons showed the existence of Giardia duodenalis assemblage A in 67 samples; G. duodenalis assemblage E in 24 samples; and inconsistent results between the two molecular markers used in a further sample. Cryptosporidium was only detected by PCR in 3 of the 274 samples; Cryptosporidium parvum was identified in two samples and Cryptosporidium ubiquitum in one sample. This study is the first performing molecular characterisation of both parasites in Peruvian alpacas, and the first report of C. ubiquitum in this host. The identification of G. duodenalis assemblage A, C. parvum and C. ubiquitum, suggests that zoonotic transmission of these enteropathogens between alpacas and humans is possible.     

Prevalence of Cryptosporidium spp. in pigs in Shanghai, China

A survey on prevalence of Cryptosporidium spp. in pigs at 12 farms in 8 suburban districts and 1 county of Shanghai was conducted under Sheather's sucrose flotation protocol and modified acid-fast stain methods from 2006 to 2009. A total of 2323 faecal samples were collected and Cryptosporidium spp. oocysts were detected in 800 samples (34.4%). Cryptosporidium was found in all 12 pig farms. Significant variations of prevalence were observed in different farms ranging from 14.1% to 90.6%. A follow-up survey on a positive pig farm for 13 consecutive months revealed that the most serious infection of Cryptosporidium spp. in pigs happened in winter and spring, and the lowest infection season was summer. Cryptosporidium spp. infection was mainly found in piglets within 2 months and no infection was found among those pigs of 90-180 days of age. The genotype analyses were carried out through PCR-RFLP and partial sequences analysis of small subunit ribosomal RNA (SSU rRNA) in some of the positive samples. Cryptosporidium suis (57/69, 82.6%), Cryptosporidium pig genotype II (6/69, 8.7%) and mixed infection of above two species/genotype (6/69, 8.7%) were found to be the main species/genotype in pigs in Shanghai area.     

Prevalence and molecular characterization of bovine Cryptosporidium isolates in India

A survey based on PCR assay of 18S SSU rRNA gene revealed a 30.2% infection with Cryptosporidium spp., out of 457 faecal samples collected from neonatal bovine calves across three different regions of India. The PCR-RFLP pattern of the gene in all the positive cases established the species as Cryptosporidium parvum. Highest prevalence was recorded in the monsoon months (37.3%) and in the calves showing acute diarrhoea (32.3%). The calves below 15 days of age were mostly affected (45.1%). The infection was more prevalent in the northern parts (35.4%) of the country than in the eastern or southern parts. Results indicated that C. parvum was the only species of Cryptosporidium prevalent in bovine calves in three different geographical regions of India.     

Giardia duodenalis and Cryptosporidium spp. in a veterinary college bovine teaching herd

In a preliminary study, we commonly identified Giardia duodenalis in adult dairy cattle from a veterinary college teaching herd. Therefore, the present study was carried out in order to better understand the potential of adult cattle to act as a source for G. duodenalis infections for students and staff at the veterinary college. Fecal samples were collected bi-weekly from this herd of adult cattle (n=30) over an 8-month period to determine the prevalence of G. duodenalis and Cryptosporidium spp. within the herd. Nested PCR followed by DNA sequencing was then performed on a subset of positive samples in order to better understand the zoonotic potential of these infections. Every cow was sampled between 11 and 18 times, depending on the date the animal joined the teaching herd. In total, 507 fecal samples were collected from 30 different cows and examined for cysts and oocysts using epifluorescence microscopy. G. duodenalis prevalence during the course of the study ranged from 37% (11/30) to 64% (18/28), with a mean of 49%. Cumulative G. duodenalis prevalence was 73% (22/30). Zoonotic G. duodenalis assemblage A genotype was identified in 43% (6/14) of the G. duodenalis-positive samples on which PCR and genetic sequencing were successfully performed. G. duodenalis assemblage E was identified in 57% (8/14) of these samples. Cryptosporidium spp. oocysts were not detected in the feces of any cows during the study period. The presence of the zoonotic G. duodenalis assemblage A in 43% of the sequenced samples indicates that there is a potential risk of infection for students and staff at this research and teaching facility, although the roles of cows as sources of giardiasis in humans remain uncertain. Furthermore, due to the large amount of feces they produce, adult cattle may serve as important sources for G. duodenalis infections in young cattle, or other animals in the facility, despite relatively low numbers of cysts excreted per gram of feces. In contrast, the results of this study indicate that this herd posed a negligible risk of transmitting Cryptosporidium parvum infections to humans.     

Prevalence and determinants of Cryptosporidium spp. infection in smallholder dairy cattle in Iringa and Tanga Regions of Tanzania

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.