A. Grinberg

Inferences about the Global Population Structures of Cryptosporidium parvum and Cryptosporidium hominis

ABSTRACT Cryptosporidium parvum and Cryptosporidium hominis are two related species of apicomplexan protozoa responsible for the majority of human cases of cryptosporidiosis. In spite of their considerable public health impact, little is known about the population structures of these species. In this study, a battery of C. parvum and C. hominis isolates from seven countries was genotyped using a nine-locus DNA subtyping scheme. To assess the existence of geographical partitions, the multilocus genotype data were mined using a cluster analysis based on the nearest-neighbor principle. Within each country, the population genetic structures were explored by combining diversity statistical tests, linkage disequilibrium, and eBURST analysis. For both parasite species, a quasi-complete phylogenetic segregation was observed among the countries. Cluster analysis accurately identified recently introduced isolates. Rather than conforming to a strict paradigm of either a clonal or a panmictic population structure, data are consistent with a flexible reproductive strategy characterized by the cooccurrence of both propagation patterns. The relative contribution of each pattern appears to vary between the regions, perhaps dependent on the prevailing ecological determinants of transmission.

Controlling the onset of natural cryptosporidiosis in calves with paromomycin sulphate

A prospective, controlled-blind field trial was conducted to evaluate the efficacy and safety of paromomycin sulphate, given for 10 days from birth, in preventing natural cryptosporidiosis in calves. In the untreated control group, oocyst shedding and diarrhoea followed the pattern that had occurred on the farm before the trial, consistent with a perinatal infection, whereas in the treated group, the prepatent period was significantly longer than in the control group (P<0.01) and oocyst shedding and diarrhoea started only after the drug was withdrawn. However, the regimen did not reduce the incidence of disease in the treated group.

Genetic Diversity and Zoonotic Potential of Cryptosporidium parvum Causing Foal Diarrhea

ABSTRACT Cryptosporidium isolates from diarrheic foals in New Zealand (n = 9) were identified as C. parvum, subtyped at two polymorphic loci, and compared with human (n = 45) and bovine (n = 8) isolates. Foal C. parvum isolates were genetically diverse, markedly similar to human and bovine isolates, and carried GP60 IIaA18G3R1 alleles, indicating a zoonotic potential.

A study of neonatal cryptosporidosis of foals in New Zealand

Abstract AIM: To assess the occurrence of Cryptosporidium oocysts in faecal specimens from foals, and investigate an outbreak of neonatal cryptosporidiosis in foals revealed in the course of the study. METHODS: Faecal specimens from foals received by a diagnostic veterinary laboratory in New Zealand between 2006 and 2007 were submitted to Massey University and tested microscopically for the presence of Cryptosporidium oocysts. The Cryptosporidium isolates in the oocyst-positive specimens were genetically identified to species level. In addition, specimen submission data from the participating laboratory for 2005–2007 were examined. In the course of the study, the identification of one Cryptosporidium-positive specimen triggered an on-farm investigation. RESULTS: Twelve faecal specimens submitted by the participating laboratory between 2006 and 2007 were tested further, and three were positive for C. parvum. Specimen submission records indicated a total of 67 faecal specimens were tested for Cryptosporidium by the participating laboratory between 2005 and 2007; 12 (18%) were positive. The on-farm investigation on a broodmare farm revealed a high incidence of neonatal diarrhoea in foals; C. parvum was the only enteropathogen found in the faeces of 4/4 affected foals examined. The diarrhoea in all those foals was self-limiting, manifesting during the second week of life, resembling foal heat diarrhoea, and accompanied by a short but intense period of shedding oocysts. CONCLUSIONS AND CLINICAL RELEVANCE: The fact that Cryptosporidium parasites were identified in 18% of faecal specimens from foals analysed for this agent in 2005–2007 by the participating laboratory indicated that infection with this agent in foals is not uncommon. Collectively, the results of this and previous studies performed in New Zealand indicate C. parvum is a cause of diarrhoea in newborn foals, potentially accounting for a proportion of cases empirically diagnosed as foal heat diarrhoea. It is therefore advisable to take precautions when handling diarrhoeic foals, until this potentially zoonotic agent is ruled out in the laboratory.

The occurrence of Cryptosporidium parvum, Campylobacter and Salmonella in newborn dairy calves in the Manawatu region of New Zealand.

Abstract AIM: To determine the occurrence of Cryptosporidium parvum oocysts, Campylobacter spp and Salmonella spp in faecal samples taken from newborn dairy calves on 24 dairy farms in the Manawatu region of New Zealand. METHODS: A cross-sectional study was conducted during the 2002 calving season. Faecal samples were collected from 185 newborn calves from a convenience sample of 24 dairy farms. The samples were tested microscopically for the presence of C. parvum oocysts, and bacteriologically for the presence of Campylobacter spp and Salmonella spp. RESULTS: Infections with C. parvum were identified in 33/156 (21.2%) calves from 10 farms. More than 106 oocysts/g (OPG) faeces were detected in calves from four farms. Campylobacter spp were isolated from 58/161 (36%) calves from 18 farms; in particular, C. jejuni subsp jejuni was isolated from 11/161 (6.8%) calves from seven farms. Salmonellae were not detected. CONCLUSIONS: Despite the short and concentrated calving pattern and the long interval between calving seasons characterising most dairy farms in New Zealand, C. parvum is widespread among calves. Campylobacter spp, especially C. jejuni, rapidly colonise the intestinal tract of newborn calves. RELEVANCE: This study provided an estimate of the ecological impact of newborn dairy calves with regard to the potentially zoonotic enteric pathogens most frequently isolated from human gastrointestinal infections in New Zealand.

Retrospective cohort study of an outbreak of cryptosporidiosis caused by a rare Cryptosporidium parvum subgenotype

The occurrence of a gastrointestinal illness among a class of 96 undergraduate veterinary students in New Zealand prompted laboratory and questionnaire-based investigations. Cryptosporidium parvum was the only enteropathogen identified in 4/7 faecal specimens analysed. The C. parvum isolates carried a rare IIa GP60 allele, indicating a point-source outbreak. The infection source could not be microbiologically traced, but the investigation suggested contact with calves during a practical class as the most likely exposure. A total of 25/80 respondents to a questionnaire were defined as cases using a clinical case definition (31% attack rate). The inferred median incubation period was 5 days (range 0-11 days), and the median illness duration was 5-6 days (range 2-23 days), corroborating previous observations in experimental cryptosporidiosis. Disease was self-limiting, characterized by abdominal discomfort, diarrhoea, and in some cases, vomiting. Originating from a rural area and having had previously handled ruminants were associated with a significant risk reduction in males. All the three students who reported chronic use of steroid inhalers for treatment of asthma were cases. This case highlighted, once again, the potential hazard for explosive outbreaks of cryptosporidiosis.

Host-shaped segregation of the Cryptosporidium parvum multilocus genotype repertoire

Cattle are among the major reservoirs of Cryptosporidium parvum in nature. However, the relative contribution of C. parvum oocysts originating from cattle to human disease burden is difficult to assess, as various transmission pathways -- including the human to human route -- can co-occur. In this study, multilocus genotype richness of representative samples of human and bovine C. parvum are compared statistically using analytical rarefaction and non-parametric taxonomic richness estimators. Results suggest that in the time and space frames underlying the analysed data, humans were infected with significantly wider spectra of C. parvum genotypes than cattle, and consequently, a significant fraction of human infections may not have originated from the regional bovine reservoirs. This study provides statistical support to the emerging idea of the existence of distinct anthroponotic C. parvum cycles that do not involve cattle.

Clinically overt infections with methicillin-resistant Staphylococcus aureus in animals in New Zealand: A pilot study

Abstract AIM: To describe clinically overt infections with methicillinresistant Staphylococcus aureus (MRSA) in animals in New Zealand, characterise clinical isolates, and track their sources. METHODS: MRSA isolates identified in 2005 and 2006 by a veterinary diagnostic laboratory were referred to Massey University for confirmation and characterisation. Clinical information was extracted from the laboratory records or obtained from referring clinicians. RESULTS: Seven MRSA isolates from animals and contact persons were characterised. All the isolates belonged to the British epidemic MRSA 15 strain (EMRSA-15). Three EMRSA-15 were isolated from post-operative infections in two dogs. An EMRSA-15 indistinguishable from the isolate recovered from one dog was isolated from the anterior nares of a healthy hospital staff member involved in the care of the animal, suggesting nosocomial transmission. Other EMRSA-15 isolates of uncertain clinical significance were isolated from the femoral head of a cat, and from a sample of cows milk. All EMRSA-15 isolates were resistant to ciprofloxacin, and four were resistant to erythromycin; the latter four isolates also exhibited inducible resistance to clindamycin. CONCLUSIONS: MRSA can cause clinically overt and difficult-to-treat infections in animals in New Zealand. The rapid emergence of EMRSA-15 as the dominant MRSA strain in humans has resulted in infection spillover to animals. CLINICAL RELEVANCE: Little is known about the incidence of clinically overt infections with MRSA in animals. The cases described here illustrate the complexities involved in the pharmacological management of EMRSA-15 infections, which is compounded by the universal resistance to β-lactams, and by the strains fluoroquinolone resistance and frequent inducible resistance to clindamycin. Such complexities indicate there is a need to develop specific empirical antimicrobial treatment strategies and antibiotic susceptibility testing protocols in countries where EMRSA-15 is dominant.

Prevalence of endemic enteropathogens of calves in New Zealand dairy farms

AIM: To conduct a country-wide prevalence study of bovine group A rotavirus, coronavirus, Cryptosporidium parvum, Salmonella spp. and enterotoxigenic K99+ Escherichia coli (K99) in calves on New Zealand dairy farms. METHODS: Faecal samples (n=1,283) were collected during the 2011 calving season from calves that were 1–5 and 9–21 days-old on 97 dairy farms, and were analysed for the presence of bovine group A rotavirus, coronavirus, Cryptosporidium and Salmonella spp., and K99. Farm-level prevalences were calculated and relationships between demographic variables and the presence of enteropathogens were examined using logistic regression models. RESULTS: Of the 97 farms, 93 (96%) had at least one sample infected with enteropathogens. The standardised farm prevalences of bovine group A rotavirus, bovine coronavirus and C. parvum were 46, 14 and 18%, respectively, in calves that were 1–5 days-old, and 57, 31 and 52%, respectively, in calves that were 9–21 days-old. The farm-level prevalence of K99 was 11% in calves that were 1–5 days-old. Salmonella spp. were found in three and four samples, from calves that were 1–5 and 9–21 days-old, respectively. No associations between explanatory variables and the presence of the enteropathogens were identified at the farm level. At the calf level, the odds of C. parvum shedding and of co-infection with any combination of pathogens were greater in calves that were 9–21 than 1–5 days-old. CONCLUSIONS AND CLINICAL RELEVANCE: This study provides epidemiological estimates of the prevalence of calves’ enteropathogens in New Zealand, which could be used for infection risk assessment or estimation of the environmental loads of pathogens shed in cattle faeces.

Risk factors for neonatal calf diarrhoea and enteropathogen shedding in New Zealand dairy farms.

Abstract To investigate the risk factors for neonatal calf diarrhoea, a cross-sectional study was conducted on 97 New Zealand dairy farms. Faecal specimens from 1283 calves were scored as liquid, semi-solid or solid, and analysed for bovine rotavirus (BRV) and coronavirus (BCV), enterotoxigenic K99+ Escherichia coli (K99), Salmonella spp. and Cryptosporidium parvum. Calf- and farm-level data were collected by means of a questionnaire and the odds of liquid faeces calculated using mixed effects logistic regression models. Among the infectious agents, only C. parvum (odds ratio [OR] = 2.6; 95% confidence interval [CI], 1.3–5.6; P = 0.02), BRV (OR = 2.7; 95% CI, 1.3–5.9; P = 0.01) and co-infection with more than one agent (compared with mono-infection: OR = 2.5; 95% CI, 1.3–4.8; P = 0.01) were associated with increased odds of liquid faeces in calves which were 9 to 21 days old. Housing of calves in open barns so exposing them to the weather was also associated with increased odds of liquid faeces compared with closed barns (OR = 2.1; 95% CI, 1.1–12.2; P = 0.03). Vaccinating cows against calf enteropathogens (OR = 0.2; 95% CI, 0.1–0.9; P = 0.03), administering waste milk (from mastitis and/or containing antibiotics; OR = 0.4; 95% CI, 0.1–0.8; P = 0.01), the sex of calves (females compared to males OR = 0.2, 95% CI, 0.07–0.7; P < 0.01), and the use of straw for bedding (OR = 0.2; 95% CI, 0.03–0.9; P = 0.03) decreased the odds of liquid faeces. Conversely, in calves that were 1 to 5 days old, only K99 was associated with liquid faeces (OR = 4.6; 95% CI, 1.2–16.1; P = 0.02). In this age group, the odds of liquid faeces were smaller on farms where females took care of the calves, compared with males (OR = 0.4; 95% CI, 0.01–0.9; P = 0.04).