Death of a neonatal lamb due to Clostridium perfringens type B in New Zealand

ABSTRACT

Case history and clinical findings: A flock of 20 sheep was kept within three paddocks on a single property. None of the animals in the flock had been vaccinated against any disease for at least three years. Abdominal bloating and haemorrhagic diarrhoea were observed in Lamb 1 at 24 hours-of-age. The lamb subsequently died within an hour of the onset of clinical signs. Lamb 2 was 3-days-old when observed to be recumbent with opisthotonus. The lamb was treated with dextrose, vitamins B1 and B12, and penicillin G, but died 4 hours later.

Pathological findings: Examination of Lamb 1 revealed markedly increased gas within the peritoneum and within dilated loops of intestine. The intestines were dark red and contained large quantities of haemorrhagic fluid. Histology of the intestines revealed peracute mucosal necrosis with minimal accompanying inflammation. The intestinal lumen contained cell debris, haemorrhage, and myriad large Gram-positive bacilli. The intestines of Lamb 2 did not appear bloated or reddened. However, multiple fibrin clots were visible within the pericardial sac. Histopathological examination revealed small foci of necrosis within the mucosa of the distal intestine. The necrotic foci were often associated with large numbers of large Gram-positive bacilli.

Immunohistochemsitry and molecular biology: Intestinal samples from Lamb 1 were processed for Clostridium perfringens immunohistochemistry, which revealed large numbers of intralesional, positively immunostained rods. Fragments corresponding to the expected sizes for genes encoding alpha, beta, and epsilon C. perfringens typing toxins were amplified by PCR from DNA extracted from formalin-fixed sections of intestine.

Diagnosis: Lamb dysentery due to C. perfringens type B.

Clinical relevance: C. perfringens bacteria have a worldwide distribution, but disease due to C. perfringens type B has only been diagnosed in a small number of countries and has never been reported in New Zealand or Australia. C. perfringens type B produce both beta toxin and epsilon toxins, therefore both haemorrhagic enteritis and systemic vascular damage can develop. As many animals are exposed to C. perfringens without developing disease, there must be additional unknown factors that resulted in disease in these particular sheep. Vaccines that specifically protect against C. perfringens type B are available and may be recommended for use in smaller non-commercial flocks, as in the present case.     

Assessment of partial resistance to powdery mildew in Chinese wheat varieties

Field trials in two cropping seasons and two locations in central China were conducted on 60 Chinese autumn‐sown wheat varieties to assess their partial resistance to powdery mildew. Mean levels of disease severity ranged from close to 0 to more than 90%. The method of inoculation and the location in which trials were conducted affected the relative performance of the varieties, but these effects were much smaller than the main effect of variety. The area under the disease progress curve was highly correlated with final disease severity, but both were poorly correlated with apparent infection rate. Disease severity was regressed against frequencies of virulence in the Blumeria graminis (syn. Erysiphe graminis) f sp. tritici populations in the trial plots. A vertical distance (D) from the mean mildew severity to the fitted line was calculated for each variety and was used to quantify partial resistance. Five of the 60 varieties, ‘Hx8541’, ‘E28547’, ‘Chuan1066’, ‘Zhe88pin6’ and ‘Lin5064’, consistently expressed relatively low levels of disease despite high frequencies of virulence in the pathogen and had consistently high D‐values. They may therefore have good levels of partial resistance.     

Study of some factors affecting the growth and development of myoga (Zingiber mioga Roscoe)

In order to optimise production of the edible flower buds produced by myoga (Zingiber mioga Roscoe) a greater understanding of the growth and development of the plant and factors influencing flower initiation and development are required. The vegetative growth phase in myoga was characterised by a distinct period of pseudostem development from the planted rhizome piece followed by an extended period of rhizome growth from both the base of pseudostems and the planted rhizome piece. The transition from pseudostem formation to the initiation of rhizome growth occurred at the same time as the dry weight of the planted rhizome piece ceased to decline. Flower bud initiation and development occurred over an extended period, beginning soon after the commencement of new rhizome growth and ending prior to foliage senescence. Flower buds were observed at the terminal meristem of first, second, third and fourth order rhizomes. Increasing temperatures stimulated both increased vegetative growth and flower bud initiation and development. Low flower bud yields recorded under conditions of low temperature were the result of reduced rate of floral initiation and not abortion of flower buds. Increased shading of plants grown under glasshouse conditions resulted in reduced flower bud yield and similarly the response resulted from decreased initiation rather than abortion.     

Extraction methods for quantitation of gentamicin residues from tissues using fluorescence polarization immunoassay

Sodium hydroxide digestion of unhomogenized kidney and skeletal muscle for 20 min at 70 degrees C was a superior method for extracting gentamicin from tissues, compared with simple homogenization, trichloroacetic acid precipitation of homogenized tissue, and sodium hydroxide digestion of homogenized tissue. Fluorescence polarization immunoassay was used to quantitate gentamicin. Sodium hydroxide digestion of unhomogenized tissue allowed for the recovery of 90.0 +/- 5.9% (means +/- SD) from renal cortex and 79.9 +/- 3.5% from skeletal muscle. The limit of sensitivity was 17.4 ng/g kidney tissue, 15.8 ng/g digested muscle, and 39.0 ng/g digested heart. The within-assay coefficient of variation (CV) at 100 ng/g kidney was 9.2%; at 500 ng/g kidney, the CV was 2.5%; and at 2000 ng/g kidney, the CV was 1.5%. The between-assay coefficient of variation was less than 7.5% for all concentrations from kidney, and the 99% confidence interval at 100 ng/g kidney was 71.7-112.4 ng gentamicin/g kidney. The within-assay coefficient of variation (CV) at 100 ng/g muscle was 15%; at 500 ng/g muscle, the CV was 2.6%; and at 2000 ng/g muscle, the CV was 2.3%. The between-assay coefficient of variation was less than 15% for all concentrations from muscle, and the 99% confidence interval at 100 ng/g muscle was 72.5-136.8 ng gentamicin/g muscle. Gentamicin-free milk could be distinguished from milk containing gentamicin concentrations of 10 ng/mL milk with 95% confidence, and from milk containing concentrations of 30 ng gentamicin/mL milk with 99% confidence. Quantitative results at or below the tolerance level can be obtained within 90 min of sample acquisition using these extraction and assay methods.     

Comparison of phenotypic characteristics of Salmonella spp isolated from healthy and ill (infected) chickens

Phenotypic characteristics of 12 paired, Salmonella serotypes isolated from healthy and ill chickens were compared. Variables compared included antibiotic resistance profiles, production of colicins and siderophores, mannose-sensitive hemagglutination of erythrocytes, resistance to serum complement, carbon source utilization, presence and transmissibility of R plasmids, and invasiveness in primary chicken kidney cell culture. Differences were found between pairs for utilization of carbon sources, mannose-sensitive hemagglutination of erythrocytes, and invasiveness in cell culture.     

Variation in the colour of broiler breast fillets in the UK

1. The normal range in colour of broiler breast muscle was assessed using an objective (instrumental) measure of colour in portioned breast fillets at a commercial processing plant. In addition, the relationship between colour and ultimate muscle pH (pH(ult)) was also examined. 2. Considerable variation in colour was evident although minimal inter-flock variation was found. The exceptions were 2 free range flocks which produced breast fillets significantly lighter and less red. 3. Extremes of colour are likely to be discriminated against at the point of purchase. 4. A clear relationship between pH(ult) and L* values was found which provides more evidence for the existence of a PSE like condition in broiler breast meat.     

Tobacco rattle virus serotypes and associated nematode vector species of Trichodoridae in the bulb-growing areas in the Netherlands

Soil samples from the coastal bulb-growing areas in the provinces of North- and South-Holland and the North-East Polder in the Netherlands were examined for trichodorid nematodes and tobacco rattle virus (TRV) serotypes. At least one of a total of eight species of Trichodoridae, of whichParatrichodorus pachydermus was most prevalent, was found in 93% of the samples from the provinces of North- and South-Holland and TRV, including four serotypes, was obtained from 49% of these samples. In the North-East Polder one of three species of trichodorids, of whichP. teres occurred most frequently, was present in 72% of the samples, and TRV of one serotype was obtained from 28% of these samples. The TRV isolates recovered from these samples reacted serologically with one of four antisera to strains of TRV. Virus transmitted byP. pachydermus reacted to the PRN-, byTrichodorus viruliferus to the RQ-, byP. teres to the N5- and byT. similis, to the TS-antiserum, respectively.     

Safety and efficacy of in ovo administration of infectious bursal disease viral vaccines

In ovo vaccination against Marek's disease virus and infectious bursal disease virus (IBDV) in commercial broilers in the United States is common. Little information exists as to the safety and efficacy of intermediate IBDV vaccines given in ovo. Experiments were initiated to determine the safety and efficacy of three commercially available live intermediate IBDV vaccines by in ovo route. Commonly used vaccines were given at 18 days of embryonation to specific-pathogen-free (SPF) broiler embryos (first and second study) or to commercial broiler embryos (third study) that had maternal antibody against IBDV. When any of the antigenic standard vaccines was given at full dose to SPF embryos, embryonic and 3-wk posthatch mortality increased. Vaccines also caused significant microscopic lesions in the bursa of Fabricius at 1 and 3 wk posthatch. In contrast, there was no adverse effect on embryonic or posthatch mortality when vaccines were given at half dose to SPF or commercial broiler embryos. However, significant microscopic lesions were evident at 1 and 3 wk posthatch in the bursae of SPF embryos given the vaccines at half dose. When vaccines were given at half dose to commercial broiler embryos, lesions were evident at 1 but not 3 wk of age. In the third study, in ovo vaccinated chickens were challenged with either a virulent standard (APHIS) or antigenic variant (variant E) IBDV virus at 3 wk of age. All vaccines produced at least 87% protection against the standard and 60% protection against the variant challenge IBDV, as measured by bursal weight to body weight ratios. This study was the first to examine the safety and efficacy of the three commonly used intermediate IBDV vaccines given in ovo in protection against standard and antigenic variant IBDV challenge viruses.