Prevalence and associated management factors of Cryptosporidium shedding in 50 Swedish dairy herds

Cryptosporidium parvum is a protozoan parasite causing diarrhoea in young calves. This cross-sectional study was performed to estimate the prevalence of Cryptosporidium infected herds in a sample of Swedish dairy herds and to identify potential risk factors associated with shedding of oocysts. Fifty dairy herds, selected by stratified random sampling, were included. The herds were visited once during the indoor seasons of 2005–2006 and 2006–2007. Faecal samples were collected from 10 calves, 10 young stock and 5 cows in each herd. Clinical observations of sampled animals and environmental status were recorded, and farmers were interviewed about management procedures. Faecal samples were cleaned by sodium chloride flotation and detection of oocysts was made by epifluorescence microscopy. Cryptosporidium parvum-like oocysts were found in 96% of the herds. Prevalence was 52% in calves, 29% in young stock and 5.6% in cows. Three two-day-old calves shed oocysts. Cryptosporidium andersoni was found in seven animals from four different herds. Factors associated with prevalence of shedders among sampled animals in a herd were age at weaning, cleaning of single calf pens, placing of young stock, system for moving young stock, and year of sampling. Factors associated with shedding in calves were age, placing of young stock, routines for moving young stock and time calf stays with the cow. The only significant factor in young stock was age. In cows, number of calves in the herd and type of farming (organic vs. conventional) affected shedding.     

The relationship of soil province to molecular characterization and phylogenetic analysis of Cryptosporidium parvum isolated from calves in Georgia

Although Cryptosporidium spp. are found throughout the world and in multiple environmental conditions, few data are available that explore the possibility of an association between specific environmental parameters and the species or strain of Cryptosporidium. This study examines the potential association between a particular Cryptosporidium species/strain found in calves and soil provinces in Georgia, USA. Necropsy cases spanning the years 1996-2002 were tested. No significant differences (P=0.962, chi(2) test of homogeneity) between numbers of positive cases were noted among soil provinces. Phylogenetic analysis of the sequences for the PCR products revealed sequence similarity of the products with Cryptosporidium parvum strain C1. Although, clinical Cryptosporidiosis in calves was not found to be affected by soil province and may be caused by a single genotype, other genotypes may be responsible for subclinical infection and warrant further investigation.     

Prevalence and age-related infection of Cryptosporidium suis, C. muris and Cryptosporidium pig genotype II in pigs on a farm complex in the Czech Republic

A total of 413 pig faecal samples were collected from pre-weaners (119), starters (131), pre-growers (123) and sows (40) from a farm with a closed breeding system segmented into two breeding complexes and a growing complex in the region of Vysočina, Czech Republic and screened for the presence of Cryptosporidium using staining methods and genotyping (SSU rRNA). Cryptosporidium oocysts were detected by microscopy in the faeces of 21.1% of the samples (87/413). Sequence analyses and RFLP identified C. suis in 44, Cryptosporidium pig genotype II in 23 and C. muris in 2 samples. No mixed infections were found.Pigs under 7 weeks of age were infected with C. suis only. Cryptosporidium pig genotype II was found in animals from 7 weeks of age. No relationship was found between diarrhoea and any Cryptosporidium infection in any of the different age groups (P < 0.05). The pre-weaned pigs shed significantly more Cryptosporidium oocysts than older pigs and it was associated with C. suis infection.     

Prevalence of Cryptosporidium spp. oocysts in cattle and wildlife in Morogoro region, Tanzania

Prevalences of Cryptosporidium spp. oocysts in cattle (n = 486) on five selected farms in Morogoro municipality and three species of herbivorous wildlife (n = 87) from Mikumi National Park, Morogoro, Tanzania, were determined using the modified Ziehl-Neelsen staining technique. Of 486 bovine faecal samples, 5.3% were positive for Cryptosporidium spp. The prevalence of Cryptosporidium was higher in calves less than 3 months of age compared to weaned calves and adults. Cryptosporidium spp. oocysts were detected in both diarrhoeic and non-diarrhoeic animals, but there was a significantly higher prevalence (P < 0.001) of oocyst shedding in diarrhoeic than in non-diarrhoeic animals. Of the 87 faecal specimens from the wildlife species, 36 were from the African buffaloes (Syncerus caffer), 25 from zebra (Equus zebra) and 26 from the wildebeest (Connochaetes gnou). Cryptosporidium spp. oocysts were detected in eight (22%) buffaloes, seven (28%) zebras and seven (27%) wildebeests. Confirmation of the diagnosis was performed using anti-Cryptosporidium monoclonal antibody specific for Cryptosporidium muris, Cryptosporidium parvum and Cryptosporidium baileyi (Pathasure Cryptosporidium test kit).     

Seroprevalence of Toxoplasma gondii and Neospora caninum in feral cats (Felis silvestris catus) in Majorca, Balearic Islands, Spain

Cryptosporidium spp. are common intestinal protozoan parasites that infect a wide range of hosts, including humans and livestock, worldwide. The objective of this study was to determine the prevalence of Cryptosporidium spp. in dairy calves in Prince Edward Island, Canada, and the potential for transmission of this parasite between dairy calves and humans. Fecal samples were collected from 183 dairy calves from 11 farms in Prince Edward Island. The prevalence of Cryptosporidium spp. infections in these animals was determined by examining for the presence of oocysts in the fecal samples, using immunofluorescence microscopy. Molecular characterization was done using a nested-PCR protocol to amplify fragments of the Cryptosporidium heat-shock protein 70 gene, followed by DNA sequencing. Ten calves (6.2%), representing 4 out of 11 farms tested, were positive for Cryptosporidium spp. DNA sequence analysis on five PCR positive samples demonstrated that Cryptosporidium parvum was the only species present in the calves tested, suggesting that there is a potential risk of zoonotic transmission between dairy calves and humans in this region.     

Genotyping of Cryptosporidium parvum isolates in bovine population in Kolkata and characterization of new bovine genotypes

Cryptosporidium infection may have adverse effect in health and production potential of cattle herd. The exact profile of Cryptosporidium infection in bovine population of India in general, particularly from Kolkata is scarce. We here report systematic investigation of clinical and genetic profiling of promiscuous Cryptosporidium infection in selected representative cattle farms from Kolkata as well as some surrounding local areas. The current study was conducted in the period of October to September, 2000–2001 with 149 diarrhoeic and non-diarrhoeic cattle of different age groups from two Government cattle farms, Harringhata Cattle Unit and Kalyani State Livestock Farm and animals raised by local farmers. Among these 149 samples, diarrhoea was recorded in 79 cases (53%) and non-diarrhoeic in 70 (46.9%). Out of 149 faecal samples screened microscopically, 32.9% from diarrhoeic faecal samples and 7.1% from healthy faecal samples revealed the presence of oocysts. Cryptosporidium genus was confirmed by DNA typing with nested PCR. The PCR-RFLP analysis was carried out for genotype identification. In course of PCR-RFLP, unique band patterns were obtained in two of our samples. The unusual RFLP products were characterized by DNA sequencing and homology analysis with other reported variants. This is the first report of identification and characterization of such a variant from the area of present investigation. Further study will be required to understand the phylogenetic origin and functional significance in virulence and morbidity of this genotype.     

Preventive and therapeutic efficacy of halofuginone-lactate against Cryptosporidium parvum in spontaneously infected calves: a centralised, randomised, double-blind, placebo-controlled study

The preventive and therapeutic efficacy of halofuginone-lactate (HFL) against Cryptosporidium parvum was evaluated in a study conducted from November 2004 to March 2005 on a dairy farm in Central Bohemia, Czech Republic, using 260 spontaneously infected calves. HFL (0.1 mg/kg/day) was administered orally for 7 days to 1-day-old and 8-day-old calves, respectively. In both treated groups the drug significantly, and in almost the same manner, decreased the intensity of diarrhoea (P<0.001) and faecal oocyst count (P<0.001) when compared to corresponding placebo groups. The only difference between both treated groups was the time of onset of symptoms of the infection. Over time, the clinical pattern of cryptosporidiosis in the animals treated at 8-14 days of age was similar to that seen in the groups receiving the placebo. In contrast, infection in the preventively treated group peaked about 10 days later but with the same intensity. The results of this study confirm the anticryptosporidial activity of HFL in calves, but show that the outcome of infection following preventive treatment is comparable to that observed in calves treated after the onset of symptoms.     

Molecular characterisation of Cryptosporidium isolates from pet reptiles

In this study, we investigated the presence of Cryptosporidium in 171 faecal samples from reptiles commonly used as pet animals. These include lizards belonging to the genera Eublepharis, Pogona, Chlamydosaurus, Hemiteconyx, Teratoscincus, Tiliqua, Iguana, and Chamaeleo, snakes of the genera Lampropeltis, Elaphe, Python, Boa and Corallus, and tortoises belonging to the genera Testudo and Kinixys. Cryptosporidium oocysts were detected by immunofluorescence using a commercially available kit and cryptosporidial DNA by amplification of a polymorphic fragment of the 18S rDNA and the HSP70 locus.Cryptosporidium was detected in 38.6% and 25.1% of the samples analysed by immunofluorescence and PCR, respectively. Molecular characterisation of the isolates confirmed that C. serpentis and C. varanii (syn. C. saurophilum) are the main species involved in infection in pet reptiles but also showed the presence of C. parvum and C. muris, as well as other species or genotypes of this parasite including the Cryptosporidium mouse genotype and Cryptosporidium tortoise genotype previously described in reptiles. In addition, a Cryptosporidium sp. was isolated from a chameleon and a python.     

Prevalence and molecular characterization of bovine coenurosis from Eastern Anatolian region of Turkey

This study was conducted to determine the prevalence and molecular characteristics of Coenurus cerebralis, the metacestode of Taenia multiceps in cattle from Erzurum province located in eastern region of Turkey between November 2009 and April 2010. Five of 1045 brains of cattle (0.47%) were found to be infected with Coenurus cysts. The characteristics and morphology of C. cerebralis were seen in all the cysts. The cyts from three infected cattle were genetically analyzed and confirmed to be T. multiceps metacestodes by NAD1 and COX1 mitochondrial gene sequence analysis. Pairwise comparison between the NAD1 sequences of the T. multiceps isolates from Erzurum and other T. multiceps isolates available in GenBank showed differences ranging from 0.6 to 2.9%, while COX1 sequences showed differences ranging from 0.2 to 2.6%. Considering the two genes, it was seen that all of the three isolates from Erzurum province were in the same group according to phylogenetic analyses. The present findings could provide a stimulus for future studies on the systematic relationships and epidemiology of lesser-known taeniid cestodes in the region, employing mitochondrial sequence data sets.     

Cryptosporidium parvum GP60 subtypes in dairy cattle from Buenos Aires,Argentina

Cryptosporidium parvum from 73 dairy calves less than two months old from Buenos Aires province (Argentina) were molecularly characterized using sequence analysis of the GP60 gene. Seventy-five sequences were obtained, and seven different subtypes were identified, all belonging to the IIa subtype family. The most common subtypes were IIaA20G1R1 (27/75), IIaA22G1R1 (16/75), and IIaA18G1R1 (13/75). Subtypes IIaA21G1R1, IIaA23G1R1, IIaA16G1R1 and IIaA19G1R1 were found sporadically. Two samples contained mixed infections with IIaA21G1R1 and IIaA22G1R1. A significant association was found between subtypes and geographic location, whereas there was no relation between subtypes and presence of diarrhea. Three of the subtypes found in this study (IIaA16G1R1, IIaA18G1R1, and IIaA19G1R1) were previously identified in humans. These findings suggest that cattle could play an important role in the transmission of cryptosporidiosis to humans in Buenos Aires province.     

Natural infection by Cryptosporidium sp., Giardia sp. and Eimeria leuckarti in three groups of equines with different handlings in Rio de Janeiro, Brazil

To detect Cryptosporidium sp., Giardia sp. and Eimeria leuckarti in horses, fecal samples were collected from three different handling horse groups from the state of Rio de Janeiro, Brazil. Group A was composed of “Mangalarga Marchador” pure breed horses, Group B was formed by horses of a Military Corporation and Group C by stray horses captured by the Center of Zoonosis Control Paulo Dacorso Filho. A total of 396 fecal samples were collected, 212 samples from Group A, 154 samples from Group B and 30 from Group C. The material was submitted to the centrifugation - flotation technique and staining by the safranin-methylene blue technique and analyzed. Oocysts of Cryptosporidium sp. were identified in 0.75% of the samples (n = 3); cysts of Giardia sp. in 0.5% (n = 2) and oocysts of E. leuckarti in 0.5% (n = 2). One case of E. leuckarti in group A and one of Cryptosporidium sp. in group B were observed. In group C were observed two cases of Cryptosporidium, two of Giardia and one of E. leuckarti,. Horses of group C were more parasitized by the three protozoans than animals from the other groups (p < 0.01). It was possible to verify that factors related to the animals, like host individual susceptibility and sanitary factors may influence the occurrence of natural infections by gastrointestinal protozoans, although the age did not have influence. This study reports, for the first time, the occurrence of Cryptosporidium sp., Giardia sp. and E. leuckarti in equines of the State of Rio de Janeiro.     

Prevalence of anti-Toxoplasma gondii and anti-Neospora caninum antibodies in sheep from Mossoró, Rio Grande do Norte, Brazil

Toxoplasma gondii is an obligate intracellular parasite with a variety of hosts, responsible for reproductive problems and economic losses in sheep flocks. Neospora caninum was recently identified and its clinical presentation in sheep is similar to that of toxoplasmosis, which can cause repeated abortions, though less frequently in this species. In order to confirm the prevalence of these agents in the city of Mossoró, Rio Grande do Norte, Brazil, 409 serum samples from adult sheep (364 females and 45 males) were tested by the indirect immunofluorescence antibody test, using cut-off point at a dilution of 1:64 and 1:50 for T. gondii and N. caninum, respectively. From the 35 properties examined, 23 (65.7%) had at least one seropositive animal for T. gondii and six (17.1%) for N. caninum. The prevalence of seropositive animals for T. gondii was 20.7% and for N. caninum 1.8%. There was no association between the presence of the agent’s antibody and gender, reports of reproductive problems and presence of dogs and/or cats in the properties. T. gondii is well distributed and N. caninum has low prevalence in sheep and in the properties of the studied region.     

Risk factors of bovine tuberculosis in cattle in rural livestock production systems of Ethiopia

This study shows a representative stratified cluster sample survey of the prevalence of comparative intradermal tuberculin test in cattle from four regions in Ethiopia. Using a cut-off for positivity of 2 mm, it assesses possible risk factors for tuberculin-positive reaction in cattle. Seventy-three villages in 24 kebeles (administrative units) were randomly selected, from which 2216 cattle from 780 owners were tested. In addition, 450 of these cattle owners were interviewed for risk factor assessment. Ninety-nine percent of the tested cattle in this rural livestock production system were traditional zebus. The individual overall prevalence of cattle bovine tuberculosis (BTB)e was 3%, with the highest found in Meskan Mareko, in Central Ethiopia (7.9%) and the lowest in Woldia, in the North East edge of the Rift Valley (1.2%). Generalised Linear Mixed Models (GLMM) with random effect on kebeles was used to analyse risk factors of cattle reactors and human tuberculosis (TB) infection. Purchase of cattle and presence of other livestock in the herd were statistically significant, with OR: 1.7, p-values of 0.03 and OR: 2, p = 0.05, respectively. Family members diagnosed with TB or showing clinical signs of extra-pulmonary TB (EPTB) were reported in 86 households (19%). None of the assessed potential risk factors of disease transmission between cattle and human (food consumption, livestock husbandry and presence of BTB-positive cattle) were statistically significant.     

Prevalence and molecular characterization of Giardia duodenalis and Cryptosporidium spp. in dairy cattle in Ontario, Canada

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.     

Phenotypic and genotypic characterization of Prototheca zopfii in a dog with enteric signs

This is a case report of enteric protothecosis caused by Prototheca zopfii in an eight-year-old male mixed breed dog with a history of chronic bloody diarrhea, loss of appetite and weight loss. Algae were isolated from rectal scrapings in defibrinated sheep blood agar and dextrose Sabouraud agar. Cytological evaluation showed the presence of globular and cylindrical organisms with a defined capsule and variable number of endospores, characteristic of the genus Prototheca, in the rectum of the animal. Scanning electron microscopy of P. zopfii strains at different development stages confirmed the diagnosis of algal infection. Molecular identification using a conserved 18S rDNA gene sequence determined that the strain belonged to genotype 2. This report describes success on treatment of canine protothecosis, diagnosed based on clinical, cytological, microbiological, scanning electron microscopy and genotypical findings.     

Detection of Fusobacterium necrophorum and Dichelobacter nodosus in lame cattle on dairy farms in New Zealand

Lameness in the dairy industry in New Zealand causes a problem in lost production, animal welfare and associated costs. To understand what bacteria may be present on the hooves of lame dairy cattle in this grass-fed system, samples were scraped from lame dairy cows and examined for the presence of Fusobacterium necrophorum (F. necrophorum) and Dichelobacter nodosus (D. nodosus) using the polymerase chain reaction (PCR). The PCR primers were designed to detect the presence of the lktA gene, which encodes a leukotoxin unique to F. necrophorum, and the fimA gene of D. nodosus. A total of 148 hoof scrapings were collected by farm staff over the period September 2005 to May 2006. F. necrophorum was detected in 79/148 of the samples, while D. nodosus was detected in 7/148 of the samples. The frequent finding of F. necrophorum within dairy herds in New Zealand is noteworthy and the occasional finding of D. nodosus on some dairy cattle suggests a possible role in both ovine and bovine hoof pathology.     

Hypodermosis in cattle slaughtered in Nigde province,Turkey<Superscript>*</Superscript>

This study was carried out to investigate the prevalence of hypodermosis in cattle between January and June 2005 in Nigde province, which is located in the middle of Turkey. A total of 1336 cattle, which were slaughtered in Nigde, were investigated for Hypoderma larvae. The 68 out of 1336 cattle (%5.08) were found positive for Hypoderma larvae. A total of 536 Hypoderma larvae were found in the skin and subcutaneous tissue of the back of infested cattle. The 489 out of 536 larvae (%91.23) were identified as Hypoderma bovis and 47 out of 536 (%8.77) as H. lineatum. Number of Hypoderma larvae counted on single infested cattle varied between 1–45 and the mean number of Hypoderma larvae per cattle was 7.88 (536/68). Hypodermosis was recorded for the first time in cattle from Nigde. ^*This study was supported by the Scientific Research Projects Unit of Nigde University (FEB 2004/07).     

Pathology of naturally occurring bovine tuberculosis in England and Wales

The aim of this study was to obtain a contemporary data set of pathology in tuberculin reactor and in-contact cattle in England and Wales. Four hundred animals (200 reactors and 200 in-contacts) from 242 farms located in 14 counties in Western England and Wales were examined. The mean number of lymph nodes (LNs) with tuberculosis (TB)-like lesions per TB-confirmed animal was 1.7 in reactors and 1.5 in in-contact animals. Tuberculous lesions in both reactor and in-contact animals were most commonly observed in the LNs of the thorax, followed by the head and abdomen, particularly the mediastinal, retropharyngeal and tracheobronchial LNs. Twenty-five reactors had macroscopic lesions in the palatine tonsils. Among TB-confirmed cattle, 27% of reactors and 9% of in-contact animals had gross TB-like lesions in the lungs, particularly in the caudal lobes. Gross lesions that were not TB-confirmed were parasitic granulomas (45%), bacterial or mycotic club-forming pyogranulomas (27%) and bacterial abscesses (23%). Diagnostic sensitivity was maximised when bacteriology and histopathology were used concurrently. Stage IV granulomas, alone or in combination with other stages, constituted 63% of lesions, while 16% of lesions were stage I/II granulomas. Caseous necrosis and calcification were common features of the granulomas encountered in natural Mycobacterium bovis infections, even with pathology limited to a small number of sites. Granulomas often covered large areas of histological sections and typically contained only small numbers of acid fast bacilli.     

Cryptosporidiosis – an occupational risk and a disregarded disease in Estonia

Background

Cases of cryptosporidiosis have not been officially reported in Estonia after the year 2000, and the disease appears to be either under-diagnosed or under-reported.

Findings

Based on a human case of cryptosporidiosis contracted during faecal sampling in dairy farms, cattle considered to be sources of infection were analysed for Cryptosporidium spp. by a modified Ziehl Neelsen technique and molecular typing. C. parvum subtype IIaA16G1R1 was detected from the human case and from calves from one of nine farms enrolled in the study providing strong circumstantial evidence of zoonotic transmission from calves to humans.

Conclusion

Cryptosporidiosis presents an occupational risk to people with cattle contact, and may also be a risk to the human population in general. Thus increased public and medical awareness is warranted.     

Prevalence and intensity of Otodectes cynotis in kittens from Thessaloniki area, Greece

From May 2007 to May 2008 we have examined by otoscopy a total number of 214 cats, aged between 0 and 6 months, brought in for their first veterinary examination to a private veterinary clinic. All cats were of urban origin. In all positive cats we performed a washing of the ear using warm paraffin oil in order to determine intensity of infection. Statistical analysis was performed using the Chi-square test. A total number of 30 cats were found to be infected with Otodectes cynotis (prevalence 14.02%). Prevalence of infection was significantly (p < 0.05) higher in cats aged between 3 and 6 months (17.58%) than in cats aged below 3 months (11.38). Intensity of infection ranged between 7 and 85 (mean intensity 35.60) mites per infected cat. The same age group of cats between 3 and 6 months had significantly (p < 0.05) higher mean intensity (47.19) compared to cats aged under 3 months (22.36). No statistical significance was found between males and females for neither prevalence nor intensity. Differences and similarities with other studies are discussed.