Health screening for a translocation of captive-reared tuatara (Sphenodon punctatus) to an island refuge

AIMS: To screen tuatara undergoing translocation from a captive crèche to an island refuge for evidence of health and known diseases, and apply basic epidemiological techniques to assess the significance of disease test results.

METHODS: Tuatara (n=353) were physically examined and samples were taken from a random selection (n=30) for estimated white cell counts, screening for haemoparasites, and culture for Salmonella, Yersinia, Aeromonas and Campylobacter spp. Direct faecal smears were carried out on-site, and faecal floats were later performed to assess levels of endoparasitism with helminths and protozoa (n=69). Modified Ziehl-Neelsen staining was used to screen faecal smears, and positive specimens were further screened using an immunofluorescence antibody (IFA) test for Cryptosporidium oocysts.

RESULTS: There was no evidence of external parasites on any of the animals examined and only one animal had a gross abnormality. All estimated white cell counts were in the range 2.8– 17.5 × 10⁹/L. No haemoparasites were observed. There were no enteric pathogens cultured, indicating the intestinal carriage of these bacteria in the tuatara was 9.4%. Of the 69 individual faecal samples examined, 12 (17%) had unidentified coccidial oocysts, 21 (30%) had nematode ova of various kinds, and 12 (17%) had intestinal carriage of motile protozoa consistent with Trichomonas spp and another unidentified organism. Nineteen (28%) tuatara had acid-fast oocysts present; however, IFA staining failed to detect any Cryptosporidium oocysts.

CONCLUSIONS: Our understanding of the diversity of gastrointestinal endoparasites affecting tuatara is inadequate as many of the parasite ova seen could not be identified. This is the first record of tuatara as a host for Trichomonas spp of protozoa in the gastrointestinal tract.     

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.     

Comparison of conventional coproscopical methods and commercial coproantigen ELISA kits for the detection of Giardia and Cryptosporidium infections in dogs and cats

Faecal samples of 270 dogs and 100 cats from 3 animal shelters in Germany were comparatively examined using conventional coproscopical methods and commercial coproantigen ELISA kits for the detection of Giardia and Cryptosporidium infections. Giardia cysts were found in 9.5% and 0% of the faecal samples in dogs and cats, respectively, examined once using the ZnCl2-NaCl flotation. However, the Giardia coproantigen ELISA (ProSpecT Giardia Microplate Assay) was positive in 29.5% and 22.4% of the samples from dogs and cats, respectively. Direct faecal smears stained with carbol fuchsin showed Cryptosporidium oocysts in one dog (0.4%) and one cat (1%). In contrast, the Cryptosporidium coproantigen ELISA (ProSpecT Cryptosporidium Microplate Assay) reacted positively in 23% of the samples from dogs and 30% of the samples from cats. Both coproantigen ELISAs were more often positive in coproscopically Giardia-negative canine faecal samples that contained Isospora burrowsi/ohioensis oocysts than in faecal samples without any parasite stage. Possible reasons for these observations are discussed.     

The prevalence of Cryptosporidium spp. oocysts in wild mammals in the Kruger National Park, South Africa

This study determined the prevalence of Cryptosporidium spp. oocysts in faecal samples from elephant (Loxodonta africana), buffalo (Syncerus caffer) and impala (Aepyceros melampus) in the Kruger National Park (KNP) and an adjacent game reserve in South Africa. Two of the study areas were in close proximity to rural communities on the western KNP boundary and the third study area was located in the centre of the KNP. Fresh stool samples (n=445) were collected and tested using an immunofluorescent antibody test (IFA) for Cryptosporidium parvum. A total of 278 of these were randomly selected (approximately 90 samples per wildlife species) and tested with the modified Ziehl Neelsen staining technique (ZN) for Cryptosporidium spp. The prevalence of Cryptosporidium spp. was highest in elephants (25.8% [95% confidence interval: 17.3, 35.9]), compared to buffalo (5.5% [1.8, 12.4]) and impala (4.3% [1.2, 10.5]). C. parvum showed similar patterns, being most prevalent in elephants (4.2% [1.5, 8.8]), compared to buffalo (1.4% [0.2, 5.1]) and impala (1.9% [0.4, 5.3]). 29 samples, including ZN positive and IFA positive samples, were retested using a real time PCR (rtPCR) technique. Of the 28 ZN-positive samples, 14 (50%) were positive with rtPCR and of the 9 IFA-positive samples 6 (67%) were confirmed positive by rtPCR. The prevalence of Cryptosporidium oocysts was significantly higher in both of the two study areas adjacent to the western KNP boundary compared to the area in the centre of the KNP (OR=3.2 [1.2, 9.0]; P=0.024). Our study demonstrates for the first time the presence of Cryptosporidium spp. in wildlife in South Africa. The transmission of this parasite between wildlife, domestic animals and humans is a plausible hypothesis and represents a potential risk for immunodeficient human populations.     

Failure to detect Salmonella species in a population of wild tuatara (Sphenodon punctatus)

AIM: To assess the prevalence of faecal excretion of Salmonella serovars by wild tuatara (Sphenodon punctatus) on Stephens Island, New Zealand. METHODS: One hundred cloacal swabs obtained as part of health-screening for the translocation of adult tuatara from Stephens Island were subjected to general aerobic culture and enrichment, and cultured specifically for Salmonella spp. RESULTS: No Salmonella spp were cultured from any of the cloacal samples, which suggests that, at the 95% confidence interval, the maximum prevalence of tuatara in the island population that were shedding Salmonella spp not detected by our sample size was 1.5%. Mixed bacteria were grown from the 70 cloacal swabs cultured aerobically. A predominant organism was evident in 30 cultures, and these were identified as Hafnia alvei type 1 (n=16) and type 2 (n=7), Corynebacterium spp (n=4), Klebsiella oxytoca (n=2), and Moraxella spp (n=1). CONCLUSIONS: The absence of intestinal carriage of Salmonella spp by the tuatara sampled in this study may indicate either lack of exposure, or an innate resistance to intestinal colonisation in tuatara. The significance of the other bacteria cultured as potential pathogens to the tuatara and as zoonotic risks is also uncertain. Wildlife managers should screen translocated reptiles for Salmonella spp, and thereby avoid exposing wild and managed populations to infection.     

Enteric infections in veal calves: a longitudinal study on four veal calf units

Forty-five calves on four veal calf units were monitored during the first four weeks after their arrival. Faecal samples were collected on alternate days and screened for the presence of rotaviruses, bovine coronavirus, Cryptosporidium oocysts, K99 positive strains of E. coli and Salmonella spp. Rotaviruses and Cryptosporidium were the most commonly detected agents (78% and 60% respectively of the calves). Bovine coronavirus was detected in the faeces of 18% of the calves, whilst K99 positive E. coli was only found in 2 samples from one calf. Salmonella spp. were not isolated from any of the 646 faecal samples examined. Shedding of rotaviruses occurred in a bimodal pattern beginning in the first week of the survey. Cryptosporidium oocysts were detected most frequently in the interval between the two peaks of rotavirus shedding. The presence of rotaviruses or Cryptosporidium oocysts in faeces was not strongly associated with scour, nor were combined infections with these agents or the cases of bovine coronavirus infection. The condition of the calves throughout the survey was generally satisfactory.     

Gastrointestinal Parasites of Stabled and Grazing Horses in Central and Northern Greece

The aim of this investigation was to compare parasites found in feces from stabled and grazing horses in Greece. For this, a total of 223 fecal samples were collected from horses from various parts of Central and Northern Greece. One hundred fifty were stabled and 73 were grazing horses, used for riding, working, or breeding. One hundred came from seven riding clubs, 73 from one breeding farm, and 50 were work horses from five farms. Parasitologic investigation was performed by applying three fecal examination techniques (Teleman's, zinc flotation, and stained smears) to detect ova, larvae, and oocysts of parasites. It was found that 77 the 223 horses (34.5%) in the study were infected with one or more parasite species. In the stabled horses, the most common parasites detected were eggs of strongyles, Strongyloides spp, Anoplocephala spp, Habronema spp, and Parascaris equorum and oocysts of Eimeria spp and Cryptosporidium spp; in the grazing horses, Anoplocephala spp. Strongyles were significantly more prevalent in the group of stabled horses than in the other group (P < .05). The results of the current study demonstrate that parasite infection is highly prevalent in horses in Greece.     

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.     

Enteric infections in veal calves: A longitudinal study on four veal calf units

Summary

Forty‐five calves on four veal calf units were monitored during the first four weeks after their arrival. Faecal samples were collected on alternate days and screened for the presence of rotaviruses, bovine coronavirus, Cryptosporidium oocysts, K99 positive strains of E. coli and Salmonella spp. Rotaviruses and Cryptosporidium were the most commonly detected agents (78% and 60% respectively of the calves). Bovine coronavirus was detected in the faeces of 18% of the calves, whilst K99 positive E. coli was only found in 2 samples from one calf. Salmonella spp. were not isolated from any of the 646 faecal samples examined. Shedding of rotaviruses occurred in a bimodal pattern beginning in the first week of the survey. Cryptosporidium oocysts were detected most frequently in the interval between the two peaks of rotavirus shedding. The presence of rotaviruses or Cryptosporidium oocysts in faeces was not strongly associated with scour, nor were combined infections with these agents or the cases of bovine coronavirus infection. The condition of the calves throughout the survey was generally satisfactory.     

Detection of Cryptosporidium oocysts in goat kid faeces: comparison of a latex agglutination test with three other conventional techniques

Detection of Cryptosporidium oocysts from goat kid faeces: comparison of a latex agglutination test with three other conventional techniques. A quantitative latex agglutination test (QLAT) with monoclonal antibodies for the detection of Cryptosporidium oocysts in faeces was compared with 3 other conventional techniques: Heine staining on faecal smears (HS) giving semi-quantitative results (scores from 1 to 5), sucrose flotation on diluted faeces (SF) with results expressed in oocysts/g of faeces (opg), direct ELISA (DE) giving qualitative results. Goat kid unconcentrated faecal samples (234) from 8 farms were processed according to the 4 techniques. Data were analyzed with Win Episcope 1.0 and Testview 1.1 softwares. The oocyst outputs ranged from 100 000 (detection limit for SF) to 200 millions opg (mean: 15.2 millions opg). A very good agreement was recorded between QLAT and HS, SF, DE: Kappa values ranged between 0.82 and 0.90. When considering the samples exhibiting oocysts (or not) as positive (or negative) using both HS and SF (n = 219), the sensitivity and specificity of QLAT were respectively 95.1 and 96.0%. The lack of sensitivity was observed in faeces harboring a few oocysts (< or = 200 000 opg, scores < or = 2) whereas the lack of specificity was only observed in 3 samples originating from the same farm. A significant correlation was calculated between the percentage of agglutination in QLAT and the number of oocysts in SF or scores in HS (Spearman correlation ranging from 0.45 to 0.48, p < 0.001). QLAT is a rapid, simple and reliable tool for routine detection of Cryptosporidium oocysts in faeces.     

Prevalence of Microsporidia, Cryptosporidium spp., and Giardia spp. in beavers (Castor canadensis) in Massachusetts.

Feces from 62 beavers (Castor canadensis) in Massachusetts were examined by fluorescence microscopy (IFA) and polymerase chain reaction (PCR) for Microsporidia species, Cryptosporidium spp., and Giardia spp. between January 2002 and December 2004. PCR-positive specimens were further examined by gene sequencing. Protist parasites were detected in 6.4% of the beavers. All were subadults and kits. Microsporidia species were not detected. Giardia spp. was detected by IFA from four beavers; Cryptosporidium spp. was also detected by IFA from two of these beavers. However, gene sequence data for the ssrRNA gene from these two Cryptosporidium spp.-positive beavers were inconclusive in identifying the species. Nucleotide sequences of the TPI, ssrRNA, and beta-giardin genes for Giardia spp. (deposited in GenBank) indicated that the four beavers were excreting Giardia duodenalis Assemblage B, the zoonotic genotype representing a potential source of waterborne Giardia spp. cysts.     

Cryptosporidium spp. in ruminants at the Lisbon Zoo.

Feces from 34 species of ruminants housed at the Lisbon Zoo was examined for Cryptosporidium spp. oocysts. Three hundred and eighty-eight samples were analyzed. Three hundred and eighty species-specific group fecal samples were collected monthly, from September 1998 until August 1999, along with eight individual specimens from eight neonates. All samples were examined by four different techniques: microscopic observation of direct and concentrated fecal smears, staining with modified Ziehl-Nielsen, immunofluorescent assay, and immunoenzymatic assay. The prevalence of infection was 3.6%. Five neonates with diarrhea were infected. Cryptosporidial oocysts were shed more frequently during winter months. Some facilities may have permitted oocysts to remain viable, possibly contributing to cryptosporidial transmission between animals.     

Evaluation of two commercial disinfectants on the viability and infectivity of Cryptosporidium parvum oocysts

Cryptosporidiosis is mainly a problem in neonatal ruminants. Not only do Cryptosporidium spp. spread ubiquitously in our environment, but the protozoa are highly resistant to harsh environmental conditions and disinfectants, and a control measure is urgently required. This study investigated the potential biocidal activity on Cryptosporidium parvum oocysts of two commercial disinfectants developed originally to be used in farms and food-processing industries. The products, containing formaldehyde and hydrogen peroxide respectively, both had some anticryptosporidial effects. The viability and infectivity of purified C. parvum oocysts exposed to both disinfectants at different concentrations and exposure times were evaluated by inclusion or exclusion of vital dye (propidium iodide), use of an excystation technique and infection of suckling mice. Viability assays showed a decrease in oocyst viability associated with an increase in exposure time for each of the concentrations used. The intensity of infection in neonatal mice was significantly lower (P<0.05) than in the control litters.     

Cryptosporidium and Giardia associated with reduced lamb carcase productivity

On two extensive sheep farms in southern Western Australia, 111 (Farm A) and 124 (Farm B) female crossbred lambs (2-6 weeks old) were randomly selected and individually identified using ear tags (a numbered tag and radio-frequency tag) at marking. On five separate occasions, faecal samples were collected and live weight, body condition score (BCS), faecal consistency score (FCS), breech fleece faecal soiling score and faecal dry matter percentage (DM%) were recorded. Lamb hot carcase weight (HCW) and dressing percentage were measured at slaughter. Faecal samples were screened by PCR for Cryptosporidium (18S rRNA, actin and 60 kDa glycoprotein [gp60] loci), Giardia duodenalis (glutamate dehydrogenase [gdh] and triosephosphate isomerise [tpi]) and Campylobacter jejuni (16S rRNA). Observation of Eimeria oocysts and faecal worm egg counts (WECs) were performed using a modified McMaster technique. The WECs were adjusted for FCS for analyses. Faecal samples were screened for patent strongylid infections using PCR (specifically ITS-2 nuclear ribosomal DNA for Teladorsagia circumcincta, Trichostrongylus spp. and Haemonchus contortus). Lambs positive for Cryptosporidium at least once had lighter HCWs by 1.25 kg (6.6%) (P=0.029) and 1.46 kg (9.7%) (P<0.001) compared to lambs never positive for Cryptosporidium for Farms A and B respectively. Similarly, dressing percentages were 1.7% (P=0.022) and 1.9% (P<0.001) lower in Cryptosporidium-positive lambs on Farms A and B respectively. Lambs positive for Giardia at least once had 0.69 kg (P<0.001) lighter HCWs and 1.7% (P<0.001) lower dressing percentages compared to lambs never positive for Giardia on Farm B only. Cryptosporidium-positive lambs at the second sampling were 4.72 (P=0.010) and 3.84 (P=0.002) times more likely to have non-pelleted faeces compared to Cryptosporidium-negative lambs for Farms A and B respectively. Breech fleece faecal soiling scores of Cryptosporidium-positive lambs were 3.36 (P=0.026) and 2.96 (P=0.047) times more likely to be moderate to severe (scores 3-5), compared to negative lambs at the second sampling for Farms A and B respectively. Live weight, growth rate and BCS were inconsistently associated with protozoa detection across different samplings and farms. Adjusted WEC was correlated positively with FCS and negatively with faecal DM%, differing between sampling occasions and farms. Campylobacter jejuni prevalence was very low (<1%). Adjusted WEC were not correlated with carcase attributes, growth rates or live weights. This study is the first to quantify productivity consequences of naturally acquired protozoa infections in lambs managed under extensive farming conditions.     

The prevalence of intestinal parasites in dogs from Prague, rural areas, and shelters of the Czech Republic

The prevalence of intestinal parasites was evaluated by examination of dog faecal samples in the Prague city centre, agricultural areas, and two shelters. The overall prevalence of parasites (i.e., protozoa and helminths, mentioned below) in Prague was 17.6%. Toxocara canis was the most common parasite, and was recovered from 6.2% of dogs, followed by Cystoisospora spp. (2.4%), Cryptosporidium spp. (1.4%), Trichuris sp. (1.1%), Taenia-type (1.0%), Giardia spp. (0.1%), Toxascaris sp. (0.9%), Dipylidium sp. (0.7%), Sarcocystis spp. (0.6%), Capillaria spp. (0.6%), Neospora/Hammondia spp. (0.5%), Ancylostoma sp. (0.4%), Uncinaria sp. (0.4%), and Spirocerca sp. (0.2%). The prevalence of infections with helminths and protozoans in two animal shelters in Prague was examined at the dog's admittance ir reception to the shelters and during housing. T. canis eggs (6.5%), Cystoisospora (4.4%), and Giardia (3.3%) cysts were the most prevalent. Significant increases in the prevalence of some parasites were found after a stay in the shelter. Giardia spp. showed an 11-fold increase in prevalence of dogs placed in the shelters for a longer time; Cryptosporidium spp. had a 7-fold increase, Capillaria spp. a 5-fold, Spirocerca sp., Neospora/Hammondia spp., and Cystoisospora spp. a 4-fold increase over dogs examined at the time of admittance to the shelter (p<0.01). Dog in rural areas were infected significantly more frequently (p<0.01) than those in Prague. In 540 faecal samples from rural areas, the overall prevalence of parasites (i.e., protozoa and helminths mentioned below) was 41.7%. The prevalence of T. canis was 13.7%, followed by Cystoisospora spp. (8.0%), Taenia spp. (3.5%), Sarcocystis spp. (3.0%), Giardia spp. (2.2%), Cryptosporidium spp. (2.0%), Trichuris sp. (1.7%), Toxascaris sp. (1.7%), Dipylidium sp. (1.3%), Neospora/Hammondia spp. (1.3%), Spirocerca sp. (1.1%), Uncinaria sp. (0.9%), Ancylostoma sp. (0.7%), and Capillaria spp. (0.6%). Examinations of dogs in urban and rural areas showed, with the exception of Trichuris sp. in Prague, a higher occurrence of nematode infection in autumn, notably T. canis (chi2>8.3, d.f.=3, p<0.04).     

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.     

The first report on Cryptosporidium suis and Cryptosporidium pig genotype II in Eurasian wild boars (Sus scrofa) (Czech Republic)

A total of 193 faecal samples of adult Eurasian wild boars were collected at 12 enclosures across the Czech Republic and examined for Cryptosporidium infection using both microscopic and molecular tools. Cryptosporidium oocysts were not detected in any of the 193 faecal samples examined using the aniline-carbol-methyl violet staining method. Thirty-two positive cases of Cryptosporidium infection were detected using either genus- or species-specific nested PCR. Mono-infection with Cryptosporidium suis and Cryptosporidium pig genotype II were found in 13 and 7 cases, respectively. Five mixed infections of C. suis and Cryptosporidium pig genotype II were detected using PCR/RFLP with genus specific primers. The number of detected mixed infections increased 2.4 fold when a species-specific PCR was employed. No other Cryptosporidium spp. was detected. Unlike cryptosporidiosis of domestic pigs, C. suis was detected as a dominant species infecting adult Eurasian wild boars. There was no association between diarrhoea and the presence of Cryptosporidium infection in the Eurasian wild boars studied. This is the first report on the Cryptosporidium infection caused by C. suis and Cryptosporidium pig genotype II in Eurasian wild boars (Sus scrofa).     

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.     

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.     

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.