Prevalence and analysis of potential risk factors for Cryptosporidium parvum infection in lambs in Zaragoza (northeastern Spain).

An epidemiologic study was carried out to investigate the prevalence of and to identify factors associated with the risk of Cryptosporidium infection in sheep in Zaragoza (northeastern Spain). Faecal samples from 583 lambs aged from 1 day to 3 months and 205 ewes older than 1 year were collected at 89 farms in the two regions of the province of Zaragoza with the highest sheep population (Zaragoza and Ejea de los Caballeros). In every sheep farm, data of the factors potentially associated with the likelihood of C. parvum infection were analysed: geographical location, season, size of herd, number of lambs in the herd at sampling time, lambing period, cleaning of lambing area and presence of diarrhoeic lambs in the farm. C. parvum oocysts were identified by using the Ziehl-Neelsen technique in 344 lambs (59%) from 75 farms (84.4%). Infected lambs ranged from less than 7 days to 90 days of age, although the percentage of animals shedding oocysts peaked at 8-14 days of age (76.2%). Statistical analysis showed that infection rates were significantly higher in lambs aged between 1 and 21 days (66.4%) than in those aged between 22 and 90 days (23%) (P<0.0001, chi(2)). Analysis of correlation between excretion of oocysts and diarrhoea revealed a relationship in all age groups and the probability of presenting diarrhoea was significantly higher for lambs shedding oocysts (86.3%) than for those which did not excrete the parasite (32.2%) (P<0.0001, chi(2)). Similarly, cryptosporidial infection rates were significantly higher in diarrhoeic (79.4%) than in non-diarrhoeic lambs (22.4%). Furthermore, infection intensity was correlated with the presence of clinical symptoms. Presence of diarrhoeic lambs in the farm was the only factor significantly associated with an increased risk of infection since the percentage of herds testing positive was significantly higher in farms with diarrhoeic lambs (91.3%) than in those without cases of neonatal diarrhoea (12.5%) (P<0.0001, chi(2)). Factors associated with a decreased risk of C. parvum infection in lambs included low numbers of lambs in the farm and cleaning of the lambing area. Additionally, lambs 8-14 days of age were less likely to be infected at the first lambing period and in spring/autumn. Cryptosporidial infection was also detected in 16 ewes (7.8%) which excreted few oocysts and without diarrhoea.

Prevalence of intestinal parasites, including Cryptosporidium parvum, in dogs in Zaragoza city, Spain.

Faecal samples from 81 dogs aged between 2 months and 13 years were collected in the small animal clinic (37 domestic dogs) and the animal shelter (44 stray dogs) located in the Faculty of Veterinary Sciences in Zaragoza city (northeast Spain) and screened for the presence of Cryptosporidium oocysts. Faeces were concentrated by the formalin-ethyl acetate method and smears of the sediment were stained by using the modified Ziehl-Neelsen technique. Cryptosporidium parvum oocysts were detected in six dogs (7.4%) aged from 2 months to 6 years. Infection was detected in both domestic (three) and stray (three) dogs and all of them excreted few oocysts (0-1 oocyst per 20 x field). No statistically significant differences in prevalence occurred between dogs younger than 6 months (11.8%) and the older dogs (6.2%). Prevalences were not significantly different between domestic (8.1%) and stray dogs (6.8%). Diarrhoea was recorded in three of the positive dogs (50%), although additional enteric parasites such as oocysts of Isospora spp. were also detected in their faeces. Nevertheless, prevalence was significantly higher in diarrhoeic (30%) versus non-diarrhoeic (4.2%) dogs (P < 0.05). Cryptosporidium was one of the parasites most frequently detected in the dogs surveyed.

Prevalence of Cryptosporidium and Giardia infections in cattle in Aragón (northeastern Spain).

Faecal samples from 554 bovines randomly selected at 30 farms in Aragón were examined to investigate the prevalence of Cryptosporidium and Giardia infections. C. parvum oocysts were identified by using the Ziehl-Neelsen modified technique in 109 (19.7%) bovines ranging from 3 days old to adults. Positive animals were found in 19 (63.3%) farms. As much as 44.4% of calves aged 3-4 days were infected, but infection rates peaked at 6-15 days of age (76.7%). Nevertheless, prevalence was also high in weanling calves aged 1.5-4 months (14%), fattening calves and heifers 4-24 months old (7.7%) and adults (17.8%). Diarrhoea was recorded in 78.6% of suckling and 29.4% of weanling calves infected by C. parvum, but it was only found to be statistically associated with infection in suckling calves (P < 0.01). All calves shedding moderate or many oocysts had diarrhoea, whereas asymptomatic infection was always correlated with few oocysts in faeces. Cryptosporidial infections were always asymptomatic in bovines older than 4 months. Giardia cysts were identified in 65 bovines (11.7%) from 16 (53.3%) of the farms surveyed. Infection rates were significantly higher in suckling (14.1%) and weanling calves (38%) than in bovines older than 4 months (2.2%) (P < 0.001). Diarrhoea was recorded in 45.5% of suckling and 10.9% of weanling calves infected by Giardia, but it was not found to be statistically associated with infection. In fact, infection rates were higher in non-diarrhoeic than in diarrhoeic calves.

Comparison of oocyst shedding and the serum immune response to Cryptosporidium parvum in cattle and pigs.

A comparison was made between oocyst shedding and the presence of specific serum IgG antibodies to Cryptosporidium parvum in 108 bovines and 90 pigs. Oocysts were detected by a commercial immunofluorescence assay in feces from 26.8% of bovines and 34.4% of pigs, whereas positive titers as determined by an indirect fluorescent antibody method were found in sera from 12.9% and 48.9% of the respective animals. Infection was significantly most frequent in suckling calves (82.7%) and weaned piglets (87.5%). By contrast, the numbers of seropositives were highest in weaned calves (17.1%) and fattening pigs (76.6%). The results of coprological and serological analysis corresponded in 65.7% of bovines and 56.7% of pigs. When used to diagnose the shedding of cryptosporidial oocysts, the detection of specific IgG antibodies had a sensitivity ranging from 10.3% (cattle) to 58.1% (pigs) and a specificity of 86.1% (cattle) and 55.9% (pigs).

Detection of Blastocystis sp. in pigs in Aragon (Spain).

Infection by Blastocystis sp. has been detected for the first time in pigs in Spain. Detection was carried out by the ethyl acetate-formalin concentration method and direct microscopy. Vacuolated cells of Blastocystis sp. were found in stool samples from 27 of 360 pigs examined (7.5%). The protozoan was present on nine of 17 pig farms studied (53%). Infected pigs ranged from 1 month old to adults, but the organism was most frequent in 1-2 month old (18.4%) and 2-6 month old pigs (15.4%). No correlation was found between Blastocystis sp. infection and occurrence of diarrhoea in pigs.

Occurrence of blastocystis sp. in cattle in Aragón, northeastern Spain.

The occurrence of Blastocystis sp. in cattle is reported herein for the first time in Spain. Stool samples from 554 bovines ranging from 3 days old to adults collected from 30 cattle farms in Aragón (northeastern Spain) were concentrated by the formalin-ethyl acetate method and examined light microscopically for the presence of gastrointestinal parasites. Vacuolated cells of Blastocystis sp. was detected in fecal specimens from 10 calves (1.8%) aged from 7 weeks to 6 months. Infected calves were detected at 3 of the 30 farms surveyed (10%). Blastocystis sp. cells were rounded or oval, with a diameter ranging from 6.25 to 15 microns. Blastocystis infection in cattle was never correlated with diarrhea, although infection intensities were generally low.