Elvira Ares-Mazás

Prevalence of and risk factors involved in the spread of neonatal bovine cryptosporidiosis in Galicia (NW Spain).

Abstract An epidemiological study was carried out on farms in Galicia (NW Spain) to investigate the prevalence of and the risk factors associated with the spread of infection by Cryptosporidium parvum in calves of less than 3 weeks of age. A total of 22 cattle farms (10 beef herds and 12 dairy herds) were visited once every 21 days between January and December 2000. A faecal sample was collected directly from the rectum of each of the 844 calves born during the study period. Each sample was studied macroscopically to establish its consistency as liquid, soft or solid, and the presence of mucus or blood noted. C. parvum oocysts were identified by direct microscopic examination and the intensity of infection established semiquantitatively as slight, moderate or severe. Of the 844 calves, 404 were found to have the parasite in their faeces, i.e. the prevalence was 47.9%. Statistical analysis of the risk factors such as general characteristics of the farm and the housing conditions of the calves, revealed three variables that significantly effect the risk of cryptosporidial infection in suckling calves: the method of cleaning, the type of flooring and the frequency of cleaning.

Batch solar disinfection inactivates oocysts of Cryptosporidium parvum and cysts of Giardia muris in drinking water

Aim:  To determine whether batch solar disinfection (SODIS) can be used to inactivate oocysts of Cryptosporidium parvum and cysts of Giardia muris in experimentally contaminated water.

Detection of oocysts and IgG antibodies to Cryptosporidium parvum in asymptomatic adult cattle

Infection by Cryptosporidium was detected in 94 (71.75%) asymptomatic adult cattle from 131 fecal samples examined microscopically. In two cases Cryptosporidium oocysts were observed which were distinctly larger (5.5-6.5 microns x 6.6-7.0 microns) than those we had seen in the majority of feces examined (4.0-4.5 microns x 4.0-4.5 microns) and these specimens were considered to be Cryptosporidium muris; it is possible that the other oocysts should be considered as Cryptosporidium parvum. The seroprevalence of IgG antibodies to Cryptosporidium was 63.35% as detected by indirect fluorescent antibody test (IFAT) and 51.41% by enzyme-linked immunosorbent assay (ELISA). In 27 cases, the presence of IgG antibodies to Cryptosporidium (as tested by IFAT and ELISA) in serum samples was correlated with oocyst excretion.

A study of cryptosporidiosis in a cohort of neonatal calves

During calving time on an experimental farm, 32 newborn calves were selected at random and monitored for infection with Cryptosporidium parvum for the first 30 days of their lives. The animals were fed pooled colostrum for 2-3 days after birth and housed in individual pens, which were washed daily using a pressure hose. Fecal smears were examined by microscopy after staining with carbol fuschin for visualization of oocysts. Oocyst shedding was scored semiquantitatively according to the average number of oocysts in 20 randomly selected fields at 1000x magnification. All the animals acquired the infection before 18 days of age. The period of maximum risk was between 9 and 12 days; 50% of the animals were infected by 9.4 days of age. It was found that the earlier the animals acquired the infection, the longer the patent period. Oocyst shedding, which did not always begin with the onset of diarrhea, lasted between 8 and 23 days (mean 12.4+/-3.3 days). Furthermore, fecal samples from 32 periparturient cows (within +/-7 days of giving birth) were filtered, concentrated and examined for oocysts using a fluorescent monoclonal antibody test, which revealed that six of the cows, although asymptomatic, were excreting C. parvum oocysts.

Giardia duodenalis and Cryptosporidium parvum infections in adult goats and their implications for neonatal kids

During the kidding season between January and April 2003, 10 farms were selected and divided into two groups of five. The farms in group A had had serious diarrhoeal illness and losses in neonatal kids the previous year, and there were Cryptosporidium parvum infections in kids associated with diarrhoea during the survey. On the farms in group B, there was no history of diarrhoeal disease the previous year and neither C parvum oocysts nor diarrhoea were detected in neonatal kids during the survey. Faecal samples were collected once from 10 adult goats aged between one and seven years on each farm. To assess more accurately the pattern of output of oocysts of C parvum and cysts of Giardia duodenalis by periparturient adult goats, one farm was selected from each group, faecal samples were collected weekly before and after kidding from 12 goats on the farm in group A and from 10 goats on the farm in group B. There was no significant difference in the prevalence of G duodenalis cysts between the group A farms (14 per cent) and the group B farms (12 per cent), and the numbers of cysts excreted ranged from 143 to 400 cysts per gram of faeces (cpg) on the group A farms and 72 to 334 cpg on the group B farms. There was a significant difference (P=0·03) in the prevalence of C parvum oocysts at the group level between the group A farms (20 per cent) and the group B farms (6 per cent). All the adult goats excreted cysts and oocysts at some date around the kidding period; the number of animals excreting cysts of G duodenalis or oocysts of C parvum increased when they gave birth, and seven to 10 times more cysts and oocysts were shed in the three weeks around kidding than in the period more than three weeks from kidding (P<0·001).

Cryptosporidium parvum in cattle, sheep and pigs in Galicia (N.W. Spain).

Infection by Cryptosporidium parvum has been detected for the first time in cattle and swine in Galicia (N.W. Spain). The organisms were also found in one of 69 sheep examined. Although in most cases the parasite was found in young diarrhoeic animals, its presence in asymptomatic mature cattle and piglets was also detected.

Herd-level risk factors for Cryptosporidium infection in dairy-goat kids in western France.

Abstract We conducted a cross-sectional study of risk factors for herd-level kid positivity for Cryptosporidium parvum oocysts in dairy-goat farms (Deux-Sèvres, western France). From January to March 2003, faeces from a convenient sample of 879 5- to 30-day-old goat kids from 60 herds were examined microscopically after staining with carbol fuschin. Oocyst shedding was scored semi-quantitatively (0 to 4+) allowing us to obtain a cumulative score per herd. Standardized questionnaires with information about management practices were collected in each farm. We found positive kids in 32 of 60 herds (53.3%) and in 142 animals out of 879 (16.2%). We used logistic regression for two risk-factor model: (1) simple positive (case: herd score ≥1+, at least one positive kid in the herd, versus control: herd score=0), (2) strongly positive (case: overall herd score ≥3+ versus control: herd score <3+). Risk factors associated with simple positive herds were period of sampling compared to the peak of births (After versus Before, OR=4.2, 95% CI 1.2, 15.3) and practice of kid grouping by age or weight (Yes versus No, OR=4.4, 95% CI 1.0, 19.1). Risk factors associated with strongly positive herds were period of investigation (February/March versus January, OR=12.7, 95% CI 2.1, 76.6), exposure to graminaceous plants in forage (OR=11.6, 95% CI 1.7, 81.0) and type of ventilation in the goat premises (Vertical versus Wind effect, OR=14.7, 95% CI 2.1, 106.1). No important association was found between kid-management practices and herd positivity. These results suggest a major role of the environment of kids during their first hours of life in the adult-goat premises regarding the transmission of C. parvum infection.

An outbreak of disease associated with cryptosporidia on a red-legged partridge (Alectoris rufa) game farm.

An outbreak of disease associated with cryptosporidia on a red-legged partridge (Alectoris rufa) game farm is described. Morbidity (diarrhoea and cough) was between 60% and 70% during the first weeks of life (4 to 25 days) and mortality was higher than 50%. The results of bacteriological and virological analyses were negative. Histological examination and antigenic diagnosis by enzyme-linked immunosorbent analysis revealed the presence of Cryptosporidium spp. in respiratory and intestinal tracts. The application of polymerase chain reaction–restriction fragment length polymorphism techniques and sequencing of a Cryptosporidium oocyst wall protein gene fragment confirmed the existence of Cryptosporidium meleagridis in faecal samples. The results obtained suggest that avian cryptosporidiosis should be included among respiratory and enteric diseases routinely tested for in farmed birds.

Cryptosporidium Contamination in Harvesting Areas of Bivalve Molluscs

Cryptosporidium contamination was evaluated in areas in Galicia (northwestern Spain) where bivalve molluscs are harvested. Galicia is the main mussel-producing region in Europe. Data were collected on water contamination of effluents that are discharged into these areas. Cryptosporidium spp. were detected by immunofluorescence microscopy and molecular methods in 71% of the river water samples (n = 7), 64% of raw sewage samples (n = 11), 50% of effluents from wastewater treatment plants (n = 16), and 29.3% of the mussel samples (Mytilus galloprovincialis, n = 184). Cryptosporidium parvum was identified in all samples of contaminated mussels, Cryptosporidium muris was found in three samples of effluent from wastewater treatment plants, and Cryptosporidium baileyi was found in a sample of raw sewage. Further studies are needed to determine the parasitological quality of water in these shellfish harvesting and recreational areas. Cryptosporidium could be a public health risk from consumption of raw or undercooked contaminated molluscs and use of contaminated waters for recreational purposes.

Treatment with β-cyclodextrin of natural Cryptosporidium parvum infections in lambs under field conditions

Following the unexpected activity of the excipient β-cyclodextrin against experimental infection by Cryptosporidium parvum in suckling mice, its efficacy in the prevention and treatment of natural infections in lambs was evaluated under field conditions. Fifty-three crossbred neonatal lambs were randomly selected for the study. Treatment consisted of oral administration of an aqueous suspension of β-cyclodextrin at a dose of 500 mg/kg of body weight. To test prophylactic efficacy, the suspension was administered at 1, 2 and 3 days of age. To evaluate therapeutic efficacy, the suspension was administered on each of the 3 days following onset of diarrhoea. Infection was monitored by daily examination of faecal samples, from birth to 30 days. The criteria studied in evaluating efficacy were: oocyst shedding, the presence of diarrhoea, and weight gain at 15 and 30 days. In the group that received prophylactic treatment with β-cyclodextrin, there were no mortalities and, compared with control lambs, there was a decrease in the number of animals infected, a longer prepatent period and notable reduction in the patent period and the duration of diarrhoea. Therapeutic treatment also reduced the patent period and the severity of diarrhoea. β-cyclodextrin was well tolerated by all of the treated animals.