Follistatin-based ligand trap ACE-083 induces localized hypertrophy of skeletal muscle with functional improvement in models of neuromuscular disease.

Skeletal muscle is under inhibitory homeostatic regulation by multiple ligands of the transforming growth factor-ß (TGFß) superfamily. Follistatin is a secreted protein that promotes muscle growth and function by sequestering these ligands extracellularly. In the present study, we evaluated the potential of ACE-083 - a locally acting, follistatin-based fusion protein - as a novel therapeutic agent for focal or asymmetric myopathies. Characterization of ACE-083 in vitro revealed its high affinity for heparin and extracellular matrix while surface plasmon resonance and cell-based assays confirmed that ACE-083 binds and potently neutralizes myostatin, activin A, activin B and growth differentiation factor 11 (GDF11). Intramuscular administration of ACE-083 caused localized, dose-dependent hypertrophy of the injected muscle in wild-type mice and mouse models of Charcot-Marie-Tooth disease (CMT) and Duchenne muscular dystrophy, with no evidence of systemic muscle effects or endocrine perturbation. Importantly, ACE-083 also increased the force of isometric contraction in situ by the injected tibialis anterior muscle in wild-type mice and disease models and increased ankle dorsiflexion torque in CMT mice. Our results demonstrate the potential of ACE-083 as a therapeutic agent for patients with CMT, muscular dystrophy and other disorders with focal or asymmetric muscle atrophy or weakness.

Volumetric MRI Study of the Brain in Fetuses with Intrauterine Cytomegalovirus Infection and Its Correlation to Neurodevelopmental Outcome.

BACKGROUND AND PURPOSE: In recent years, effort has been made to study 3D biometry as a method for fetal brain assessment. In this study, we aimed to compare brain volumes of fetuses with cytomegalovirus infection and noninfected controls. Also, we wanted to assess whether there is a correlation to their neurodevelopmental outcome as observed after several years. MATERIALS AND METHODS: A retrospective cohort study examined MR imaging brain scans of 42 fetuses (at 30-34 weeks' gestational age) that were diagnosed with intrauterine cytomegalovirus infection. Volumetric measurements of 6 structures were assessed using a semiautomated designated program and were compared with a control group of 50 fetuses. Data collected included prenatal history and MR imaging and sonographic and neurodevelopmental follow-up. RESULTS: We found that all brain volumes measured were smaller in the cytomegalovirus-infected group and that there was a correlation between smaller cerebellar volume and lower Vineland II Adaptive Behavior Scales questionnaire scores, especially in the fields of daily living and communication skills. CONCLUSIONS: In this study, we found that brain volumes are affected by intrauterine cytomegalovirus infection and that it has a developmental prognostic meaning. Such information, which should be supported by further research, may help clinicians further analyze imaging data to treat and make a better assessment of these fetuses.

Molecular detection of Bartonella coopersplainsensis and B. henselae in rats from New Zealand.

AIM To identify Bartonella spp. in rats from New Zealand using molecular methods. METHODS DNA was extracted from the spleens of 143 black rats (Rattus rattus) captured in the Tongariro National Park, New Zealand. PCR was performed using Bartonella genus-specific primers amplifying segments of the 16S-23S rRNA internal transcribed spacer and citrate synthase (gltA) and beta subunit of the RNA polymerase (rpoB) genes. PCR products were sequenced and compared online with sequences stored in the database of the National Center for Biotechnology Information of the United States of America. RESULTS DNA sequences matching Bartonella coopersplainsensis and B. henselae were detected in samples from 22/143 (15.4%) and 3/143 (2.1%) rats, respectively. Co-occurrence of B. coopersplainsensis and B. henselae sequences was observed in the sample from one rat. CONCLUSIONS AND CLINICAL RELEVANCE Gram-negative fastidious bacteria belonging to the genus Bartonella are associated with a range of human diseases. Rodents play an important role as reservoirs of a broad range of Bartonella species. To our knowledge, this is the first report of a molecular detection of Bartonella spp. DNA in rodents from New Zealand, and the first identification of B. henselae DNA in rats, worldwide. Whereas the public health significance of B. coopersplainsensis remains undefined, B. henselae is the agent of cat scratch disease, and the presence of this bacterium in rats may have public health implications. Our results are preliminary and additional analyses of larger samples, preferably by bacterial culture, would provide more information on the prevalence and diversity of Bartonella spp., in particular B. henselae, in rats.

The Effect of Urine Concentration and pH on the Growth of Escherichia Coli in Canine Urine In Vitro.

BACKGROUND: Lower urinary tract infections are common in dogs, and Escherichia coli is the most common bacterial pathogen isolated. The literature has conflicting evidence regarding the inhibitory effects of urine concentration and pH on E. coli growth. HYPOTHESIS/OBJECTIVES: To determine the effect of different pH and urine concentrations on E. coli growth in vitro. ANIMALS: Voided urine samples from 10 apparently healthy spayed female dogs were used. METHODS: A matrix of 9 urine specific gravity (USG; 1.010, 1.020, and 1.030) and pH (5.5, 7.0, and 8.5) combinations was prepared by diluting and titrating filtered voided urine samples. Three E. coli isolates were obtained from urine of female dogs with signs of lower urinary tract infection and cultured at different urine pH and USG combinations in wells of a microtiter plate. The number of E. coli colony-forming units (CFU) per mL of urine was calculated after aerobic incubation of the urine at 37°C for 18 hours, and statistically compared. RESULTS: Significant differences were identified in the mean log CFU/mL among different combinations of pH and USG. The lowest log CFU/mL were observed in alkaline concentrated urine (pH 8.5 and USG 1.030). CONCLUSIONS AND CLINICAL IMPORTANCE: Escherichia coli in vitro growth was higher in neutral to acidic and diluted urine compared to alkaline and concentrated urine. The impact of non-alkalizing diluting diets on the incidence of E. coli lower urinary tract infections should be further explored.

Characterisation of extended-spectrum ß-lactamase and AmpC ß-lactamase-producing Enterobacteriaceae isolated from companion animals in New Zealand.

AIMS: To assess the occurrence of, and characterise, extended-spectrum ß-lactamase (ESBL) and AmpC ß-lactamase (AmpC)-producing Enterobacteriaceae isolated by veterinary diagnostic laboratories from infection sites in companion animals in New Zealand. METHODS: Selected Enterobacteriaceae isolates were submitted by seven New Zealand veterinary diagnostic laboratories. They were isolated from infection sites in companion animals between June 2012 and June 2013, and were resistant to amoxicillin-clavulanic acid, fluoroquinolones, or any combination of two or more antimicrobials. Based on disk diffusion test results, the isolates were phenotypically categorised according to production of ESBL and AmpC. Genes for ESBL and AmpC production were amplified by PCR and sequenced. Escherichia coli isolates were also typed by multilocus sequence typing. RESULTS: A total of 115 isolates matching the inclusion criteria were obtained from the participating laboratories, of which 74 (64%) originated from dogs and 29 (25%) from cats. Seven bacterial species were identified, of which E. coli was the most common (87/115, 76%). Of the 115 isolates, 10 (9%) expressed the ESBL phenotype, 43 (37%) the AmpC phenotype, and seven (6%) both ESBL and AmpC phenotypes. Of the 60 ESBL and AmpC-producing isolates, 36 (60%) were E. coli. Amongst these isolates, 27/60 (45%) were classified as multidrug resistant, compared with 15/55 (27%) non-ESBL or AmpC-producing isolates (p<0.01). Ninety five isolates were resistant to amoxicillin-clavulanic acid and 58 (61%) of these were ESBL or AmpC-producing. The predominant ESBL genes were blaCTX-M-14 and blaCTX-M-15, and the dominant plasmid-encoded AmpC gene was blaCMY-2. Thirty-eight E. coli multilocus sequence types (ST) were identified, and the most prevalent were ST12 (12/89, 13%), ST131 (6/89, 7%) and ST648 (6/89, 7%). ESBL and AmpC-producing isolates accounted for 35/1,082 (3.2%) of the Enterobacteriaceae isolated by one laboratory network over the study period. CONCLUSIONS AND CLINICAL RELEVANCE: ESBL and AmpC-producing Enterobacteriaceae were associated with clinical infections in companion animals in New Zealand, and were often multidrug resistant. In this study, these organisms accounted for <5% of all Enterobacteriaceae isolated from infection sites by one laboratory network, but their prevalence among isolates resistant to amoxicillin-clavulanic acid was 61%. Therefore routine secondary testing for ESBL and AmpC production by Enterobacteriaceae that are resistant to amoxicillin-clavulanic acid in primary testing could improve the accuracy of definitive antimicrobial therapy in companion animals in New Zealand.

Characterisation of methicillin-resistant Staphylococcus aureus clinical isolates from animals in New Zealand, 2012-2013, and subclinical colonisation in dogs and cats in Auckland.

AIMS: To characterise methicillin-resistant Staphylococcus aureus (MRSA) isolates from infection sites in animals in New Zealand and assess the prevalence of subclinical MRSA colonisation in dogs and cats attending veterinary clinics in Auckland. METHODS: MRSA isolates from clinical specimens obtained by the main New Zealand veterinary diagnostic laboratories between June 2012 and June 2013, were genotypically characterised by DNA microarray hybridisation analysis and spa typing. In addition, nasal or perineal skin swabs collected from a cross-sectional sample of dogs (n=361) and cats (n=225) attending 29 veterinary clinics in Auckland during the same period were analysed for MRSA by culture. RESULTS: Eight MRSA clinical isolates were submitted for characterisation by the participating laboratories. The isolates originated from five dogs, including two isolates from the same dog, one foal, and one isolate had no identification of the source. The strain-types identified were AK3 (ST-5 SCCmecIV t045; n=1), USA500 (ST8 SCCmecIV t064; n=1), WSPP (ST30 SCCmecIV t019; n=1), Rhine Hesse (ST5 SCCmecII t002; n=2), and EMRSA-15 (ST22 SCCmecIV t032; n=3). No MRSA were isolated from 586 cultured swabs. Methicillin-susceptible S. aureus were detected in 9/257 (3.5%) swabs and non-aureus staphylococci in 22/257 (8.5%) swabs. The estimated true MRSA subclinical colonisation prevalence was 0%, with an upper 95% CI boundary of 1.9% for cats and 1.4% for dogs. CONCLUSIONS: The modest number of MRSA isolates submitted for this study by the participating laboratories suggests clinical MRSA infection in animals in New Zealand continues to be sporadic. The wide variety of strain-types found mirrored the evolving strain-type diversity observed in humans. We cannot rule out bias due to the non-random sampling of dogs and cats, but the apparent colonisation prevalence of 0% was consistent with the low prevalence of subclinical colonisation in humans in New Zealand. These similarities indicate the epidemiology of animal and human MRSA infections are linked. CLINICAL RELEVANCE: In the last decade, the prevalence of human MRSA infections in New Zealand has steadily increased. This is the second published study of MRSA in animals in New Zealand. The results indicate clinical MRSA infection in animals remains sporadic, but the diversification of the strain-types may pose new therapeutic challenges to veterinarians, due to their diverse resistome.

Subclinical Bacteriuria in Older Cats and its Association with Survival.

BACKGROUND: Bacterial urinary tract infections are uncommon in cats in general but the prevalence increases to 29% in older cats with comorbidities (Veterinary Clinical Pathology 2008, 37, 317; Journal of Feline Medicine & Surgery 2007, 9, 124; Veterinary Microbiology 2009, 136, 130). Frequently, the infections are subclinical. The clinical relevance of subclinical bacteriuria (SB) is uncertain, and the optimal treatment requires clarification. OBJECTIVE: Prospective, observational study to: (i) identify the prevalence and incidence count of SB in older (≥7 years), nonazotemic cats, (ii) evaluate specific risk factors for SB, and (iii) investigate the potential relationship between untreated SB and survival. ANIMALS: Sixty-seven, nonazotemic cats were tested on 5 occasions over 3 years. METHODS: Urine samples were obtained by cystocentesis for quantitative urine culture and blood samples for measurement of serum creatinine concentration. Episodes of SB were not treated. Serum creatinine concentration, body weight, urine specific gravity, sex, and age were evaluated as potential risk factors for a positive urine culture. The association between urine culture results and survival was evaluated with Cox's proportional hazard model. RESULTS: A total of 256 urine samples was obtained. The prevalence of SB varied between 10 and 13%, and incident infections were uncommon. Female cats were 21 times more likely to have a positive urine culture than were male cats (odds ratio [OR], 21.2; confidence interval [CI], 4.1-110; P = .00028). Subclinical bacteriuria was not significantly associated with survival. CONCLUSION AND CLINICAL IMPORTANCE: Subclinical bacteriuria is common in nonazotemic, older cats. Although antimicrobial treatment was withheld, the presence of SB was not adversely associated with survival.

The ability of four strains of Streptococcus uberis to induce clinical mastitis after intramammary inoculation in lactating cows.

AIM: To compare the ability of four strains of Streptococcus uberis at two doses to induce clinical mastitis in lactating dairy cows after intramammary inoculation in order to evaluate their usefulness for future experimental infection models. MATERIALS AND METHODS: Four field strains of S. uberis (26LB, S418, and S523 and SR115) were obtained from cows with clinical mastitis in the Wairarapa and Waikato regions of New Zealand. Twenty-four crossbred lactating cows, with no history of mastitis and absence of major pathogens following culture of milk samples, were randomly allocated to four groups (one per strain) of six cows. Each cow was infused (Day 0) in one quarter with approximately 10(4) cfu and in the contralateral quarter with approximately 10(6) cfu of the same strain. The other two quarters remained unchallenged. All four quarters were then inspected for signs of clinical mastitis, by palpation and observation of the foremilk, twice daily from Days 0-9, and composite milk samples were collected from Days 0-8 for analysis of somatic cell counts (SCC). Quarters were treated with penicillin when clinical mastitis was observed. Duplicate milk samples were collected and cultured on presentation of each clinical case and on Day 4 from challenged quarters with no clinical signs. RESULTS: Clinical mastitis was diagnosed in 26/48 (54%) challenged quarters. Challenge with strain S418 resulted in more cases of mastitis (12/12 quarters) than strains SR115 (7/12), 26LB (6/12) or S523 (1/12), and the mean interval from challenge to first diagnosis of mastitis was shorter for S418 than the other strains (p<0.001). The proportion of quarters from which S. uberis could be isolated after challenge was less for strain 26LB (1/6) than SR115 (6/7) (p<0.05), and SCC following challenge was lower for strain S523 than the other strains (p<0.05). CONCLUSIONS: There were significant differences between the strains in the proportion of quarters developing clinical mastitis, the interval to mastitis onset, SCC following challenge and the proportion of clinical cases from which S. uberis could be isolated. These results illustrate the difference in the ability of S. uberis strains to cause mastitis and the severity of the infections caused. CLINICAL RELEVANCE: Experimental challenge models can be used to compare infectivity and pathogenicity of different strains of mastitis-causing bacteria, the efficacy of pharmaceutical products and host-responses in a cost-effective manner.

Susceptibility to antimicrobials of mastitis-causing Staphylococcus aureus, Streptococcus uberis and Str. dysgalactiae from New Zealand and the USA as assessed by the disk diffusion test.

OBJECTIVE: To compare the antimicrobial susceptibility patterns of three common mastitis pathogens (Staphylococcus aureus, Streptococcus uberis and Str. dysgalactiae) isolated from milk samples from New Zealand and the USA. METHODS: A total of 182 S. aureus, 126 Str. uberis and 89 Str. dysgalactiae isolates from New Zealand (107, 106 and 41, respectively) and the USA (75, 20 and 48, respectively) were assessed using the disk diffusion test. RESULTS: Susceptibility varied among the bacterial species. All isolates were susceptible to the amoxicillin-clavulanic acid combination. Resistance to lincomycin was most frequent (susceptibility of 8.6%) across all species. Non-susceptible (i.e. resistant or intermediate) isolates of S. aureus were identified for the three non-isoxazolyl penicillins (amoxicillin, ampicillin and penicillin: 20.6% and 36.0%) and lincomycin (99.9% and 94.6%) for NZ and the USA, respectively. Resistance to erythromycin (5.3%) and tetracyclines (6.7%) was detected only in isolates from the USA. There were differences in susceptibility between Str. uberis and Str. dysgalactiae; all streptococcal isolates demonstrated resistance to aminoglycosides (neomycin 52.4% and streptomycin 27.9%) and enrofloxacin (28%). Resistance of Str. dysgalactiae to tetracycline was almost 100.0% and to oxytetracycline 89.9%. CONCLUSION AND CLINICAL RELEVANCE: Most of the isolates tested were susceptible to most of the antimicrobials commonly used for treatment of bovine mastitis, with the exception of the lincosamides. Susceptibility to a selected class-representative antimicrobial and at the genus level should be interpreted with caution. Differences between NZ and the USA confirm the value of national surveys to determine the susceptibility patterns of mastitis pathogens.

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

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).

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.

Utility of halofuginone lactate for the prevention of natural cryptosporidiosis of calves, in the presence of co-infection with rotavirus and Salmonella Typhimurium.

Halofuginone lactate (HL) is registered in several countries for the prevention of calf cryptosporidiosis, but the compound's utility in the presence of co-infection with other enteropathogens is not well understood. We performed a randomized controlled field trial of the efficacy of HL for the prevention of natural calf cryptosporidiosis, in the presence of co-infection with rotavirus and Salmonella Typhimurium. Newborn calves on one farm were sequentially enrolled and allocated to a full dose (n=15), half dose (n=15), or a placebo control group (n=15), using a randomized block design. The Cryptosporidium oocysts in fecal specimens collected on Days 6, 8, 10, 14 and 20 were counted and the severity of the diarrhea was assessed using fecal consistency scores (solid, semisolid, or liquid). The oocyst numbers and fecal consistency scores were statistically compared between the groups. Ninety one percent of the calves shed Cryptosporidium parvum oocysts during the trial. The full dose group had a longer prepatent period than the control group, but no statistical difference in the number of oocysts was identified between the groups after controlling for the effects of sex and breed. The fecal consistency scores and mortality rates did not differ between the groups. These results indicated that the anti-Cryptosporidium activity and clinical benefit of HL were limited. It is concluded that in order to maximize the clinical efficacy of HL in the field, diagnostic efforts should aim to rule out the presence of other enteropathogens.

Extensive intra-host genetic diversity uncovered in Cryptosporidium parvum using Next Generation Sequencing.

The theory about the Cryptosporidium life cycle predicts genetic diversity of sporozoites within the host. Nevertheless, the Cryptosporidium intra-host genetic diversity is difficult to study using conventional Sanger sequencing or electrophoretic resolution of amplicons, due to the methods' inability to resolve mixtures of templates. We analysed the within-isolate genetic diversity of two Cryptosporidium parvum isolates sharing common descent, by combining the use of Next Generation Sequencing and cloning of PCR amplicons with database searches. The analysis focused on the single-copy 70 kDa heat shock protein (HSP70) and the 60kDa surface glycoprotein (gp60) genes, which allowed any diversity to be ascribed to the presence of a heterogeneous population of sporozoites. The results indicated an unprecedented intra-host genetic diversity, with two HSP70 and 10 gp60 alleles in these isolates, in spite of the initial resolution of one allele per locus using Sanger sequencing. At both loci, the predominant alleles were those initially identified by Sanger sequencing. A significant (p<0.01) overrepresentation of gp60 alleles previously reported in New Zealand was observed. These results further our understanding of the genetic structure of C. parvum populations, and expose the limitations of the use of non-axenic isolates as operational taxonomic units of genetic studies of cryptosporidiosis.

Cardiovascular responses to lower body negative pressure before and after 4 h of head-down bed rest and seated control in men and women.

Cardiovascular deconditioning after a 4-h head-down bed rest (HDBR) might be a consequence of the time of day relative to pre-HDBR testing, or simply 4 h of confinement and inactivity rather than the posture change. Ten men and 11 women were studied during lower body negative pressure (LBNP) before and after 4-h HDBR and 4-h seated posture (SEAT) as a control for time of day and physical inactivity effects to test the hypotheses that cardiovascular deconditioning was a consequence of the HDBR posture, and that women would have a greater deconditioning response. Following HDBR, men and women had lower blood volume, higher heart rate with a greater increase during LBNP, a greater decrease of stroke volume during LBNP, lower central venous pressure, smaller inferior vena cava diameter, higher portal vein resistance index with a greater increase during LBNP, but lower forearm vascular resistance, lower norepinephrine, and lower renin. Women had lower vasopressin and men had higher vasopressin after HDBR, and women had lower pelvic impedance and men higher pelvic impedance. Following SEAT, brachial vascular resistance was reduced, thoracic impedance was elevated, the reduction of central venous pressure during LBNP was changed, women had higher angiotensin II whereas men had lower levels, and pelvic impedance increased in women and decreased in men. Cardiovascular deconditioning was greater after 4-h HDBR than after SEAT. Women and men had similar responses for most cardiovascular variables in the present study that tested the responses to LBNP after short-duration HDBR compared with a control condition.

Efficacy of a novel internal dry period teat sealant containing 0.5% chlorhexidine against experimental challenge with Streptococcus uberis in dairy cattle.

The incidence of clinical mastitis and infection status at calving was assessed in quarters treated with 1 of 2 internal teat sealants at the time of dry off. Two contralateral quarters per cow (n=63 cows) were treated with a sealant that contained 0.5% chlorhexidine; the other quarters were treated with a commercial teat sealant. Ten cows were untreated (controls). On d 2, 4, and 16 after dry off, cows were challenged with Streptococcus uberis S210 strain. Cows were examined daily for 34 d after drying off and cases of clinical mastitis were recorded. Milk samples were collected for culture from any quarters that developed clinical mastitis during the first 34 d after drying-off and from all quarters on d -5 and 0 relative to treatment and at the first and twentieth milking after calving. The incidence of clinical mastitis during the examination period was lower in treated quarters (n=7/252; 1.5%; lower incidence for those treated with chlorhexidine-containing teat sealant n=3/126; 1.2%) than in untreated quarters (n=13/40; 26.8%). The protection against intramammary infection after calving, adjusted for the effect of cow, was higher in quarters treated with the novel teat sealant (89/105; 15.2%; 95% CI=9.6-23.4) than in those treated with the commercial teat sealant (71/104; 31.7%; 95% CI=23.5-41.3) and untreated controls (6/28; 78.6%; 95% CI=59.8-90.0), respectively. Quarters treated with teat sealants were less likely to have an intramammary infection after calving and had a lower incidence of clinical mastitis during the early dry period than did untreated controls in this challenge study.

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.

A study of neonatal cryptosporidiosis of foals in New Zealand.

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.

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

AIM: To describe clinically overt infections with methicillin resistant 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 cow's milk. AlleMRSA-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 spill over 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 beta-lactams, and by the strain's 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.

Genetic diversity and zoonotic potential of Cryptosporidium parvum causing foal diarrhea.

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.

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.