Cryptosporidium spp. in diarrheic calves less than 30 days old from farms across Northern Ireland were examined over a year period by microscopic, genotyping, and subtyping techniques to characterize the transmission dynamics. Cryptosporidium oocysts were detected in 291 of 779 (37.4%) animals. The prevalence rates of rotavirus, coronavirus, and Escherichia coli K99+ were lower as seen in 242 of 806 (30.0%), 46/806 (5.7%), and 16/421 (3.8%) of animals, respectively. Of the 224 Cryptosporidium-positive specimens available for molecular analysis, Cryptosporidium parvum was identified in 213 (95.1%) specimens, Cryptosporidium bovis in eight (3.6%), and Cryptosporidium deer-like genotype in three (1.3%). Sequence analysis of the 60-kDa glycoprotein gene identified 16 IIa subtypes and a new subtype family, with 120 of the 216 (55.6%) positive specimens having the subtype IIaA18G3R1. Eight of the IIa subtypes were previously seen in humans in Northern Ireland. Several subtypes were temporally or geographically unique. The genetic diversity in calves in Northern Ireland was much greater than that reported from other areas. This work demonstrates the utility of genotyping and subtyping tools in characterizing the transmission of Cryptosporidium spp. in calves and humans.
A method is presented for the detection of the nitrofuran, furazolidone, in porcine tissue. Following methanol-buffer extraction of the tissue, liquid partitioning, and solid-phase clean-up, samples are analysed by using thermospray LC-MS monitoring the positive ion m/z 243 with filament-assisted ionization. The LOD is 1 microgram kg-1. The assay is used to investigate the depletion of furazolidone from tissue and sample stability post mortem. It is necessary to snap-freeze samples by immersion in liquid nitrogen immediately upon collection in order to improve the stability of residues in tissue.
The effects of level of concentrate feeding in late gestation on feed intake, milk yield, milk composition, and fertility in the subsequent lactation were evaluated in a randomized block design experiment involving 60 cows. Grass silage was offered ad libitum for the last 28 d of gestation either as the sole diet (OC) or supplemented with 5 kg/d of concentrates (5C). Following calving, the cows were offered the same grass silages supplemented with 7 kg/d of concentrates. For treatments OC and 5C, total dry matter intakes were 9.28 and 11.03 kg/d of dry matter, respectively, during the last 4 wk of gestation. During wk 1 to 12 of the subsequent lactation, treatment 5C increased milk fat concentration but did not alter feed intake, milk yield, or protein concentration relative to treatment OC. Treatment 5C increased the interval to first progesterone rise and the number of services per conception relative to treatment OC. Cow parity, BF depth assessed at d 28 before parturition, and treatment provided the best fit relationships for the yields of fat and fat plus protein (R2 relationships = 0.65 and 0.64, respectively) during wk 1 to 4 of lactation. It was concluded that, other than milk fat concentration, supplementation with additional concentrates in late gestation did not alter milk yield or composition and dairy cow fertility. Furthermore, despite the very large differences in cow characteristics at d 28 before parturition, there was no evidence of any interaction between treatment and specific cow characteristics on animal performance in the first 12 wk of lactation.
Mature Fasciola hepatica of the triclabendazole-resistant Sligo isolate were incubated in vitro with 10 microg/ml of the sulphoxide metabolite of compound alpha [5-chloro-2-methylthio-6-(1-naphthyloxy)-H-benzimidazole]; the metabolite will be referred to as alpha.SO. Changes resulting from drug treatment were examined by scanning electron microscopy (SEM), transmission electron microscopy (TEM) and tubulin immunocytochemistry (ICC). SEM revealed that disruption to the tegumental surface mainly took the form of swelling and blebbing. Extensive spine loss occurred on the ventral surface of the oral cone, and sloughing of the tegument was observed along the lateral margins of the fluke. Examination of sections from the anterior mid-body region at the TEM level revealed that treatment with alpha.SO led to swelling of the basal infolds and mitochondria within the tegumental syncytium; also, accumulations of secretory bodies beneath the apical plasma membrane. The tegumental cell bodies contained swollen mitochondria and cisternae of granular endoplasmic reticulum, but few Golgi complexes were observed. An increase in T2 secretory bodies was observed, whilst in the T1 tegumental cells, the T1 secretory bodies had decreased in number. Immunocytochemical (ICC) studies showed that incubation with alpha.SO, ABZ.SO and TCBZ.SO did not cause significant changes to the distribution of tubulin within the tegumental syncytium of the Sligo isolate. In contrast, alpha.SO, ABZ.SO and TCBZ.SO caused severe disruption to tubulin organization within the syncytial layer of the TCBZ-susceptible Cullompton isolate. The EM results confirm that compound alpha is a fasciolicide capable of disrupting the tegument of mature TCBZ-resistant F. hepatica; however, this was not accompanied by any change in tubulin immunoreactivity.
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