A Retrospective Cohort Study of an Outbreak of Cryptosporidiosis among Veterinary Students.

An outbreak of gastrointestinal illness occurred among a cohort of 56 veterinary technology and 100 veterinary science students at Massey University over an eight-week period in 2013. This coincided with calving in New Zealand's seasonal dairy farming system and a time when calves with diarrhoea are commonly seen by veterinarians. Laboratory and epidemiological investigations were instigated by MidCentral Public Health Service (MCPHS) in conjunction with the Institute of Veterinary, Animal and Biomedical Sciences (IVABS) at Massey University. Eighty students responded to a questionnaire of which 19 met the case definition, a 24% attack rate. Faecal specimens from seven students contained Cryptosporidium oocysts and Cryptosporidium parvum IIa A18G3R1 was identified from one of the specimens. The inferred median incubation period was five days (range 1-12 days). All of the cases were self-limiting, characterized by diarrhoea, abdominal cramps, and in some cases vomiting, headache, and fever. Having contact with calves with diarrhoea was significantly associated with increased adjusted odds of being a case (OR 10.61, 95% CI 1.87-108.29 for one week of contact; OR 55.05, 95% CI 3.80-1931.18 for two weeks of contact). Outbreaks of cryptosporidiosis had occurred previously among veterinary students at Massey University, but the extremely high infectivity of C. parvum resulted in student illness despite enhanced hygiene precautions.

Insecticidal activity of bio-oil from the pyrolysis of straw from Brassica spp.

Agricultural crop residues can be converted through thermochemical pyrolysis to bio-oil, a sustainable source of biofuel and biochemicals. The pyrolysis bio-oil is known to contain many chemicals, some of which have insecticidal activity and can be a potential source of value-added pest control products. Brassicacae crops, cabbage, broccoli, and mustards, contain glucosinolates and isocyanates, compounds with recognized anti-herbivore activity. In Canada, canola Brassica napus straw is available from over 6 000 000 ha and mustard Brassica carinata and Brassica juncea straw is available from 200 000 ha. The straw can be converted by microbial lignocellulosic enzymes as a substrate for bioethanol production but can also be converted to bio-oil by thermochemical means. Straw from all three species was pyrolyzed, and the insecticidal components in the bio-oil were isolated by bioassay-guided solvent fractionation. Of particular interest were the mustard straw bio-oil aqueous fractions with insecticidal and feeding repellent activity to Colorado potato beetle larvae. Aqueous fractions further analyzed for active compounds were found not to contain many of the undesirable phenol compounds, which were previously found in other bio-oils seen in the dichloromethane (DCM) and ethyl acetate (EA) solvent phases of the present study. Identified within the most polar fractions were hexadecanoic and octadecanoic fatty acids, indicating that separation of these compounds during bio-oil production may provide a source of effective insecticidal compounds.

Migration behaviour of discontinuous buffers in capillary electrophoresis during protein enrichment.

Capillary electrophoresis (CE) is not only an effective separation technique, but can also serve as a sample preparation tool for enrichment and purification at sub-microliter sample volumes. Our approach is based on the use of a discontinuous buffer system consisting of an acid and a base (acetate and ammonium). Proteins and/or peptides with isoelectric points between the pH values of these two buffers will become stacked at the neutralization reaction boundary (NRB). To understand the mechanism of the NRB formation and the electrophoretic migration of various ions during the enrichment, we performed experiments using myoglobin and mesityl oxide to reveal the ion migration patterns at the buffer junction, and utilized Simul 5 to computer simulate the process. The simulated results closely resembled the experimental data, and together, they effectively revealed the characteristics of the discontinuous buffers. Importantly, the discovery allowed the manipulation of NRB behaviours by controlling the discontinuous buffer composition. To illustrate this, the removal of urea as an unwanted background molecule from the enriched protein sample was achieved based on the acquired information.

Removal of sample background buffering ions and myoglobin enrichment via a pH junction created by discontinuous buffers in capillary electrophoresis.

Traditional CE sample stacking is ineffective for samples containing a high concentration of salt and/or buffer. We recently reported the use of a discontinuous buffer system for protein enrichment that was applicable to samples containing millimolar concentrations of salt. In this paper, the technique was investigated for samples containing unwanted buffering ions, including TRIS, MES, and phosphate, which are commonly used in biological sample preparation. Using myoglobin as a model protein, the results demonstrated that background buffering ions can be effectively removed or separated from the enriched protein. The key is to use either the acid or the base of the discontinuous buffers to adjust the pH of the sample, such that the net charge of the unwanted buffering ions is near-zero. The successful isolation and enrichment of myoglobin from up to 100 mM TRIS and 50 mM MES was demonstrated. The enrichment factors remained at approximately 200. Removal of phosphate was more challenging because its net charge was anionic in both the acid and the base of the discontinuous buffers. The enrichment was only achievable up to 30 mM of sodium phosphate, the enrichment factors observed were significantly lower, below 50, and the process was delayed due to the higher ionic strength resulted from phosphate. The migration of phosphate during enrichment was studied using a UV-absorbing analogue, phenyl phosphate. In addition, Simul 5.0 was used to simulate the discontinuous buffers in the absence and presence of TRIS and phosphate. The stimulated TRIS and phosphate concentration profiles were generally in agreement with the experimental results. The simulation also provided a better understanding on the effect of phosphate on the formation of the pH junction.

In-capillary protein enrichment and removal of nonbuffering salts using capillary electrophoresis with discontinuous buffers.

Salt is abundant in biological samples and can cause problems in capillary electrophoresis (CE) due to excessive Joule heating and electrodispersion. Desalting with solid phase minibeds is currently most compatible with the small sample volumes of CE. They are however difficult to prepare and suffer from poor bed-to-bed reproducibility. Alternatively, enrichment of proteins and peptides was developed using CE, by trapping them at their isoelectric points with a discontinuous buffer of mismatched pH. Ionic salts, such as sodium chloride, do not possess isoelectric points and therefore are not retained by the discontinuous buffer. In this work, the removal of ionic salt during protein enrichment using CE with discontinuous buffers was investigated. Nonbuffering ions were found to electromigrate through the pH junction without disrupting the enrichment process and were eventually removed from the capillary. Mass spectral data obtained from the enriched and desalted sample confirmed a significant signal enhancement. Finally, a strong acid was introduced to remove the pH junction and thus facilitated a subsequent capillary zone electrophoresis separation. An integrated procedure of enrichment, desalting, and separation was demonstrated on a mixture of three protein standards.