Effect of colostrum quantity and quality on neonatal calf diarrhoea due to Cryptosporidium spp. infection.

This study was conducted to assess the effect of colostrum quality and quantity on Cryptosporidium spp. calf diarrhoea in an intensive dairy cattle farm in Greece. Faecal samples were collected from 100 dairy calves randomly selected and born during all 4 seasons (March 2015 to May 2016) of the year. In total, 71% of the selected calves were positive for Cryptosporidium spp. oocysts. The statistical analysis revealed influence of colostrum quality on faecal score. Linear regression showed that the colostrum quantity during the first day of life was negatively associated with the number of Cryptosporidium spp. oocysts in faeces. During multivariable analysis, the variables representing the quality of colostrum and the season of the calf's birth were identified as confounders. Cryptosporidium spp. is a common pathogen participating in neonatal calf diarrhoea. Colostrum management and season influence the number of Cryprosporidium spp. oocysts and faecal consistency. The above findings demonstrate novel risk factors that should be included in the strategic approaches to control cryptosporidiosis in newborn calves.

Two-Year Monitoring of Water Samples from Dam of Iskar and the Black Sea, Bulgaria, by Molecular Analysis: Focus on Mycobacterium spp.

The coast of the Bulgarian Black Sea is a popular summer holiday destination. The Dam of Iskar is the largest artificial dam in Bulgaria, with a capacity of 675 million m3. It is the main source of tap water for the capital Sofia and for irrigating the surrounding valley. There is a close relationship between the quality of aquatic ecosystems and human health as many infections are waterborne. Rapid molecular methods for the analysis of highly pathogenic bacteria have been developed for monitoring quality. Mycobacterial species can be isolated from waste, surface, recreational, ground and tap waters and human pathogenicity of nontuberculose mycobacteria (NTM) is well recognized. The objective of our study was to perform molecular analysis for key-pathogens, with a focus on mycobacteria, in water samples collected from the Black Sea and the Dam of Iskar. In a two year period, 38 water samples were collected-24 from the Dam of Iskar and 14 from the Black Sea coastal zone. Fifty liter water samples were concentrated by ultrafiltration. Molecular analysis for 15 pathogens, including all species of genus Mycobacterium was performed. Our results showed presence of Vibrio spp. in the Black Sea. Rotavirus A was also identified in four samples from the Dam of Iskar. Toxigenic Escherichia coli was present in both locations, based on markers for stx1 and stx2 genes. No detectable amounts of Cryptosporidium were detected in either location using immunomagnetic separation and fluorescence microscopy. Furthermore, mass spectrometry analyses did not detect key cyanobacterial toxins. On the basis of the results obtained we can conclude that for the period 2012-2014 no Mycobacterium species were present in the water samples. During the study period no cases of waterborne infections were reported.

[Effect of radix sophorae flavescentis (RSF) mixture on mast cells in jejunal mucosa of mice infected by Cryptosporidium].

OBJECTIVE: To elucidate the role of mast cells (MC) activation in the jejunal mucous membrane in the pathogenesis of cryptosporidiosis (CPS) and explore the mechanism of prevention and treatment of radix sophorae flavescetis(RSF) mixture on CPS. METHODS: A total of 30 healthy male BALB/c mice were randomly divided into a normal control group, CPS model control group and RSF mixture experimental group. The mice of CPS model were inoculated intragastrically with 1 x 10(5) Cryptosporidium oocyst (CSO). The mice in the RSF mixture experimental group were treated with inoculation of RSF mixture (0.2 ml doses) twice one week for three weeks continuously after CPS models were established. Pathological changes of the jejunal mucosa membrane were observed by a light microscope. The MCs were stained by toluidine blue, the number of mast cells was recorded and the changes of degranulation were observed. RESULTS: The HE staining showed inflammatory pathological changes in the jejunal mucosa membrane of the CPS model control group. After three-week treatment of RSF mixture, the small intestine epithelium was integrated on the whole. The toluidine blue stain showed the number of mast cell in submucosa and muscular layer of the jejunal mucous membrane increased significantly in the model control group (12.80 +/- 0.84) compared with those of the normal control group (1.60 +/- 0.89) (P < 0.01) and an obvious degranulation was seen in the CPS model control group. The number of mast cells of the mice in the RSF mixture experimental group decreased significantly (P < 0.01) and the number (2.00 +/- 0.71) and morphous were closed to the normal after administration for three weeks. CONCLUSIONS: MC activation is involved in the intestinal inflammatory response caused by Cryptosporidium. RSF mixture could decline the number of MC, inhibit the activation and degranulation of MC in the jejunal mucosa membrane of CPS mice to reduce inflammation and repair the damaged intestinal mucosa, which may realize the purpose of treatment of CPS.

Removal of Cryptosporidium and polystyrene microspheres from swimming pool water with sand, cartridge, and precoat filters.

Cryptosporidium has caused the majority of waterborne disease outbreaks in treated recreational water venues in the USA for many years running. This research project evaluated some common US swimming pool filters for removing Cryptosporidium oocysts, 5-µm diameter polystyrene microspheres, and 1-µm diameter polystyrene microspheres. A 946 L hot tub with interchangeable sand, cartridge, and precoat filters was used at room temperature for this research. Simulated pool water for each experiment was created from Charlotte, NC (USA) tap water supplemented with alkalinity, hardness, chlorine, and a mixture of artificial sweat and urine. Precoat (i.e., diatomaceous earth and perlite) filters demonstrated pathogen removal efficiencies of 2.3 to 4.4 log (or 99.4-99.996%). However, sand and cartridge filters had average Cryptosporidium removals of 0.19 log (36%) or less. The combined low filter removal efficiencies of sand and cartridge filters along with the chlorine-resistant properties of Cryptosporidium oocysts could indicate a regulatory gap warranting further attention and having significant implications on the protection of public health in recreational water facilities. The 5-µm microspheres were a good surrogate for Cryptosporidium oocysts in this study and hold promise for use in future research projects, field trials, and/or product testing on swimming pool filters.

Determining potential indicators of Cryptosporidium oocysts throughout the wastewater treatment process.

Most research on wastewater treatment efficiency compliance focuses on physicochemical and microbial indicators; however, very little emphasis has been placed so far on determining suitable indicator organisms to predict the discharge level of pathogens from treatment plants. In this study, raw wastewater, activated sludge, and the resulting final effluents and biosolids in four municipal wastewater treatment plants (WWTPs A, B, C and D) were seasonally investigated for human-virulent water-borne pathogens Cryptosporidium parvum/hominis and Giardia duodenalis, and microsporidia (e.g. Encephalitozoon hellem, E. intestinalis, and Enterocytozoon bieneusi) between 2008 and 2009. A suite of potential microbial indicators for human-virulent protozoa and microsporidia was also determined. A combination of multiple fluorescent in situ hybridization and immunofluorescent antibody assays were applied to detect Cryptosporidium oocysts, Giardia cysts, and microsporidian spores. Escherichia coli, enterococci and Clostridium perfringens spores were cultivated in selective media. Positive correlations were found between the abundance of enterococci and E. coli and abundance of Cryptosporidium oocysts (r(s) > 0.47, p < 0.01) and Giardia cysts (r(s) > 0.44, p < 0.01) at WWTPs A-D. Cryptosporidium perfringens spores were positively correlated to Cryptosporidium oocysts (r(s) = 0.40, p < 0.01) and Giardia cysts (r(s) = 0.46, p < 0.01). There was a strong positive correlation between abundance of Giardia cysts and that of Cryptosporidium oocysts (r(s) > 0.89, p < 0.01). To sum up, a suite of faecal indicator bacteria can be used as indicators for the presence of Cryptosporidium oocysts and Giardia cysts in these activated-sludge systems (WWTPs A, B and C). Overall, Giardia duodenalis was noted to be the best Cryptosporidium indicator for human health in the community-based influent wastewater and throughout the treatment process.

Cryptosporidiosis in 20 alpaca crias.

CASE DESCRIPTION: 20 alpaca crias (13 females and 7 males) were examined for diarrhea (n=20), weight loss (15), and poor appetite (5). Fourteen crias were between 8 and 18 days of age at time of admission. CLINICAL FINDINGS: Cryptosporidiosis was diagnosed in all crias. Common biochemical abnormalities included acidemia, hyperlactemia, azotemia, and hyperglycemia and increases in aspartate transaminase and gamma-glutamyltransferase activities. Serum sodium and chloride concentrations were high or low. Other potential gastrointestinal tract pathogens were identified in only 7 crias. TREATMENT AND OUTCOME: Supportive care was instituted, including i.v. administration of fluids with partial parenteral administration of nutrients (n=19 crias), antimicrobials (19), supplemental orally administered nutrients (11), administration of plasma (10), and insulin treatment (9). Other palliative treatments used by attending clinicians were sucralfate, flunixin meglumine, vitamin A/D/E/B complex, antiparasitic agents, antidiarrheal agents, and azithromycin. Three crias with inadequate urine production and severe azotemia were treated with furosemide administered i.v. as a bolus or as a constant-rate infusion. Treatment resulted in a successful outcome in 16 of 20 crias. Weight loss and refractory azotemia were common in nonsurvivors but not in surviving crias. CLINICAL RELEVANCE: Findings suggested that Cryptosporidium spp may be a diarrheal pathogen of unweaned alpaca crias that may be more widespread than has been recognized and can become endemic on some farms. Metabolic derangements were unpredictable and should be determined by biochemical analysis before fluid and electrolyte replacement is initiated. Cryptosporidiosis has zoonotic potential, and the infection can be self-limiting in alpacas receiving supportive treatment.

Pelleted organo-mineral fertilisers from composted pig slurry solids, animal wastes and spent mushroom compost for amenity grasslands.

In Ireland, conversion of biodegradable farm wastes such as pig manure spent mushroom compost and poultry litter wastes to pelletised fertilisers is a desirable option for farmers. In this paper, results obtained from the composting of pig waste solids (20% w/w) blended with other locally available biodegradable wastes comprising poultry litter (26% w/w), spent mushroom compost (26% w/w), cocoa husks (18% w/w) and moistened shredded paper (10% w/w) are presented. The resulting 6-mo old 'mature' composts had a nutrient content of 2.3% total N, 1.6% P and 3.1% K, too 'low' for direct use as an agricultural fertiliser. Formulations incorporating dried blood or feather meal amendments enriched the organic N-content, reduced the moisture in mature compost mixtures and aided the granulation process. Inclusion of mineral supplements viz., sulphate of ammonia, rock phosphate and sulphate of potash, yielded slow release fertilisers with nutrient N:P:K ratios of 10:3:6 and 3:5:10 that were suited for amenity grasslands such as golf courses for spring or summer application and autumn dressing, respectively. Rigorous microbiological tests carried out throughout the composting, processing and pelletising phases indicated that the formulated organo-mineral fertilisers were free of vegetative bacterial pathogens.

Water quality monitoring: the basis for watershed management in the Oldman River Basin, Canada.

The Oldman River flows 440 km from its headwaters in south-western Alberta, through mountains, foothills and plains into the South Saskatchewan River. Peak flows occur in May and June. Three major reservoirs, together with more than a dozen other structures, supply water to nine irrigation districts and other water users in the Oldman basin. Human activity in the basin includes forestry, recreation, oil and gas development, and agriculture, including a large number of confined livestock feeding operations. Based on the perception of basin residents that water quality was declining and of human health concern, the Oldman River Basin Water Quality Initiative was formed in 1997 to address the concerns. There was limited factual information, and at the time there was a desire for finger pointing. Results (1998-2002) show that mainstem water quality remains good whereas tributary water quality is more of a challenge. Key variables of concern are nutrients, bacteria and pesticides. Point source discharges are better understood and better regulated, whereas non-point source runoff requires more attention. Recent data on Cryptosporidium and Giardia species are providing benefit for focusing watershed management activities. The water quality data collected is providing a foundation to implement community-supported urban and rural better management practices to improve water quality.

Centralised versus decentralised sewage systems: a comparison of pathogen and nutrient loads released into Sydney's drinking water catchments.

Data collected from centralised and decentralised sewage treatment plants throughout Sydney's drinking water catchments was used to calculate the relative catchment loads of Cryptosporidium, enteric viruses, nitrogen and phosphorus for an initial screening assessment. Loads were assessed at median and 90 percentile values for expected and worst-cases scenarios. The expected scenario in the Sydney drinking water catchments is that decentralised systems (servicing 32,800 people) provide similar total loads to centralised systems (serving 70% of the catchment population) for total phosphorus (37,090 kg x y(-1)), Cryptosporidium (10(11) oocysts x y(-1)) and enteric viruses (9.1 x 10(13) y(-1)), but higher loads of total nitrogen (237,610 vs. 136,740 kg x y(-1)). Decentralised systems, however, were predicted to have higher loads in the worst-case scenario with 620,620 kg x y(-1) TN, 82,040 kg x y(-1) TP, 7.3 x 10(13) Cryptosporidium oocysts x y(-1) and 9 x 10(15) enteric viruses per year. Greater load variability was experienced with decentralised systems, which presumably reflects less reliability in their current operation and maintenance. Overall, catchment water quality is therefore not only affected by sewage disposal methods, but also failure issues. Decentralised system disposal to land may afford a degree of mitigation that can be enhanced, if the degree of failure is reduced.

Further studies on fecal parasites in antiquity.

This is a continuation study of the survival of antigenic material over the centuries using mummified human remains from the Andean area of South America. The fluorescent antibody kit from Meridian Diagnostics (Cincinnati, OH) was used to identify some Cryptosporidium species and Giardia species found in feces from the intestines of mummies 500 to 3,000 years old. The specimens that were positive by direct visualization using fluorescent antibody were then tested with an enzyme-linked immunosorbent assay reaction using a Meridian kit just released on the market. Since all of the feces used were formed, it would seem that the organisms found were from carriers rather than active cases of disease. Similar fecal specimens were shown to harbor antigens from Helicobacter pylori almost 3,000 years old.

Effect of a whey protein concentrate used as a colostrum substitute or supplement on calf immunity, weight gain, and health.

The efficacy of a whey protein concentrate was evaluated as a colostrum substitute or supplement in two experiments using four groups of 29 calves. In Experiment 1, calves were fed either 2 L of pooled colostrum (group 1) or 500 g of whey protein concentrate (group 2). A mean total of 123.6 and 17.7 g of Ig was fed to calves in groups 1 and 2, respectively. Mean serum IgG, total protein, and globulin concentrations and Ig antibody activities to Escherichia coli K99 and rotavirus were significantly higher for calves in group 1 at 24 to 36 h and at 3 wk of age. Weight gain from birth to 3 wk of age was significantly lower for calves in group 2. The incidence of diarrhea was high but not different between treatments. The mortality rate (0 to 3 wk) was significantly higher for calves in group 2 (27.6%) than for calves in group 1 (3.4%). In Experiment 2, calves were fed either 2 L of pooled colostrum (group 3) or a solution of 1 L of pooled colostrum plus 500 g of whey protein concentrate (group 4). A mean total of 117.2 and 69.1 g of Ig was fed to calves in groups 3 and 4, respectively. Absorption rate of IgG was significantly lower for calves in group 4. Mean serum IgG, total protein, and globulin concentrations and Ig antibody activities to E. coli K99 and rotavirus were significantly higher for calves in group 3 at 24 to 36 h and at 3 wk of age. Mortality rate, BW gain, and incidence of diarrhea did not differ significantly between groups.

Effects of housing and colostrum feeding on the prevalence of selected infectious organisms in feces of Jersey calves.

Neonatal Jersey calves (n = 96) were used to evaluate effects of housing (individual hutches or wooden pens in a barn) and colostrum feeding (calves were separated from the dam and fed 2 L of colostrum in nipple-bottles or allowed to nurse the dam for 3 d) on the prevalence of selected organisms in feces. Prevalence of Cryptosporidium and Eimeria were reduced, and prevalence of rotavirus tended to be reduced, when calves were housed in hutches. Prevalence of coronavirus was unaffected by treatment. Weekly prevalence of Giardia was increased when calves were left to nurse the dam for 3 d. Mean prevalence of Cryptosporidia (wk 1 to 4), Eimeria (wk 4 to 5), Giardia, rotavirus, and coronavirus (wk 1 to 5) were 34.7, 20.6, 27.1, 15.8, and 4.9%, respectively. Escherichia coli (K99 positive) were observed in 3 of 174 samples cultured. Methods of housing and colostrum feeding affected acquisition of enteropathogens in this study.

Bovine colostrum immunoglobulin concentrate for cryptosporidiosis in AIDS.

Lactobin-R is a commercial hyperimmune bovine colostrum with potent anticryptosporidial activity. It was administered to a 4 year old child with AIDS and severe diarrhoea associated with cryptosporidiosis. There was significant clinical improvement in the diarrhoea and permanent elimination of the parasite from the gut as assessed through serial jejunal biopsy and stool specimens.

Rotavirus and Cryptosporidium shedding in dairy calf feces and its relationship to colostrum immune transfer.

Twenty Holstein calves received 2 L of colostrum twice within 12 h after birth; the first feeding occurred within 2 h of parturition. The increase in adsorption efficiency was related to the gamma globulin provided in the first colostrum feeding. Absorption efficiency ranged from 2.4 to 46.1%. The number of sheddings of rotavirus and Cryptosporidium by the calves during their first 4 wk of life was associated with serum gamma globulin concentration 24 h after birth and absorption efficiency. Absorption efficiency and body weight combined accounted for 60.4% of the variation in the number of sheddings; heavier calves shed more than lighter calves. During the first 4 wk, calves that shed more frequently gained less weight; weight gain was also associated with serum gamma globulin levels 24 h after birth. Colostrum composition varied between quarters of the same cow. Total protein and gamma globulin content of colostrum from the rear quarters was higher than from the front quarters. The association between number of sheddings and absorption efficiency suggest that calves should not be fed colostrum containing less than 9 g/100 ml of total protein.