In vitro and in vivo efficacy of alpha-cyclodextrin for treatment of experimental cryptosporidiosis

The efficacy of alpha-cyclodextrin against infection by Cryptosporidium parvum was evaluated using in vitro and in vivo models. Cyclodextrins are water-soluble cyclic hexamers of glucose units with hydrophobic cavities capable of solubilizing lipophiles and are widely used as drug excipients in the pharmaceutical industry. The viability of purified C. parvum oocysts, exposed for 30, 60, 90, 120 min and 24h to different concentrations of alpha-cyclodextrin (2.5, 5, 7.5, 10, 12.5 and 15%), was evaluated by inclusion or exclusion of two fluorogenic vital dyes and by an excystation technique. Preventive and curative efficacies against cryptosporidial infections, at different doses (2.5 and 5%) and regimes of administration of alpha-cyclodextrin, were determined in an experimental neonatal mice model. Results of the viability assay showed a decrease in oocyst viability that was associated with an increase in exposure time, for each of the concentrations used. Moreover, a high proportion of nonviable oocysts (81%) was observed when C. parvum oocysts were exposed to alpha-cyclodextrin (2.5%) for 24h. The intensity of infection, determined 7 days post-inoculation by examination of intestinal homogenates, was significantly lower (P<0.05) than in the control litters, for all the assays carried out with alpha-cyclodextrin. Only 38.8% of the animals became infected when the alpha-cyclodextrin solution (5%) was administered 2h before inoculated oocysts, and every 24h at 1 and 2 days post-inoculation.     

Characterization of Cryptosporidium spp.--recent developments and future needs

Cryptosporidia, widely distributed protozoan parasites of vertebrates have recently attracted increasing interest due to several serious waterborne outbreaks, the life-threatening nature of infection in immunocompromised patients, and the realization of economic losses caused by these pathogens in livestock. Genetic polymorphism within Cryptosporidium spp. is being detected at a continuously growing rate, owing to the widespread use of modern molecular techniques. The aim of this paper is to review the current status of taxonomy, genotyping, molecular phylogeny, and characterization of cryptosporidia, and to highlight the need for polyphasic typing, i.e. an integrated approach comprising standardized morphologic, biologic, and molecular methods for describing Cryptosporidium species and isolates, and for establishing "virtual" reference strains.     

Effect of two thermal regimes on the muscle growth dynamics of sea bass larvae,Dicentrarchus labrax L

Muscle growth was studied in larvae of sea bass, Dicentrarchus labrax L., reared at two temperatures: real ambient temperature ( congruent with 15 degrees C during vitelline phase and increased gradually) and 19 degrees C from fertilization until the end of larval development. Muscle cellularity, body length and body weight were measured. Early temperature influenced larval development and so, pre-larval phase finished earlier at 19 degrees C than at ambient temperature (4 and 6 days, respectively). Temperature also affected muscle growth such that at hatching and at mouth opening hypertrophy of muscle fibres was greater at 19 degrees C (P < 0.05), whereas hyperplasia was similar in both groups. After 25 days, the cross-sectional area of the white muscle was greater at 19 degrees C (P < 0.05), which was mainly associated with a higher proliferation of new white muscle fibres. At this stage the body length was also higher at 19 degrees C. Metamorphosis finished earlier in fish reared at 19 degrees C (52 days) than at natural temperature (82 days). At this developmental stage body length and cross-sectional area of the myotome were similar in both groups. However, muscle cellularity differed between groups. Thus, hypertrophy of muscle fibres was higher in fish reared at ambient temperature (P < 0.05), whereas proliferation of new muscle fibres was higher at 19 degrees C (P > 0.05).     

Arterial vascularization of the pineal gland in the fetus of Zavot-bred cattle

This study aimed at revealing arterial vascularization of the pineal gland of the Zavot-bred foetus. Twenty foetuses, regardless of their sex, at the age of 2-7 months were used. Coloured-latex was injected by way of both the right and left common carotid arteries. Then, dissection was performed and vessels nourishing the pineal gland were documented. The pineal gland is vascularized by a number of 2-5 central rami. A small vessel arising from each of the central rami in two foetuses (10%) was shown anastomosing with a branch of the cranial cerebral artery, which advances in cranio-caudal direction in the callosal groove. Hence, anastomoses were observed between several sub-branches of each caudal cerebral and cranial cerebellar arteries.     

Bronchopneumonia and polyarthritis due to Mycoplasma bovis in a calf

This case report describes gross lesions and histopathological findings in a 3-months-old calf originating from a feedlot with approximately 400 cattle. In this animal and additional 14 cattle of similar age, which were kept together in the same stable, swollen joints had occurred suddenly. The examination of this calf showed that a severe polyarthritis induced by haematogenous spread of Mycoplasma bovis following bronchopenumonia was present, which was characterised by necrotising lesions of the joint capsules and severe cartilage erosions.     

Hepatozoon spp.: report of some cases in dogs in Brasília, Brazil

Canine hepatozoonosis is a disease caused by the tick-borne protozoan Hepatozoon spp. It has been reported in the United States, southern Europe, the Middle East, Africa and the Far East. In Brazil, canine hepatozoonosis is an emerging protozoal tick-borne disease, and is characterized by distinct clinical signs. The objective of this study was to analyze the laboratory findings of some hepatozoonosis cases in dogs in Brasília, Brazil, and their clinical signs. The animals of this experiment showed low parasitemia, similar to H. americanum, but the clinical signs presented were similar to H. canis. According to our observations and in agreement with O'Dwyer et al. [Vet. Parasitol. 94 (2001) 143], the Brazilian Hepatozoon appears more to resemble the species found in the eastern Hemisphere than with H. americanum of North America, or could be caused by a new species. Our data revealed that hepatozoonosis could be considered endemic in Brasília.     

Epidemiological study on clinical bovine dermatophilosis in northern Ethiopia

A total of 5586 indigenous and cross breed cattle were examined for clinical dermatophilosis, and 292 (5.22%) were positive. In indigenous cattle, statistically discernible (p < 0.05) difference was recorded between the prevalence in wet (5.5%) and dry (4.0%) seasons. During both seasons, infestation with A. Variegatum significantly (p < 0.05, chi 2 = 59.9) influenced clinical dermatophilosis. A. variegatum-infested cattle were at 7x higher risk (OR, Odds Ratio = 6.8) of acquiring the disease than non-infested ones. Logistic regression analysis of the overall prevalence showed a significant difference (p < 0.05) between prevalence in indigenous and cross breeds; the crossbreeds being at about 3x higher risk (OR = 2.68). Improved management reduced the prevalence in cross breeds from 15.1% in dry to 8.6% in wet season suggesting the significance of management in the epidemiology of dermatophilosis. The disease occurrence was also significantly (p < 0.05, chi 2 = 133.05) affected by agro-climatic (altitude) variation suggesting the impact of rainfall, humidity, vegetation and vector population that varied with altitude. Age and gender had no influence (p > 0.05) on the disease. Our result showed that season of the year, breed, management, infestation with A. variegatum and agro climatic difference are important risk factors influencing the disease pattern in the region. A tick control strategy and a better and adequate management particularly when keeping cross breeds are recommended.     

Histological lesions in vascular tissues of bovine herpes virus type 4-infected rabbits

The gamma-herpes virus bovine herpes virus type 4 (BoHV-4) is distributed worldwide in cattle populations with unknown pathogenicity. Bovine endothelial cells were recently shown to be susceptible to BoHV-4 infection in vitro and this virus accelerated the cholesterol-induced atherosclerotic process in rabbits. In this study, the in vivo effect of BoHV-4 on cardiovascular tissue was investigated by intravenous infection of rabbits fed a cholesterol free diet. Inflammatory lesions of vascular tissue in aortic and valvular endothelial cells, and smooth muscle cells were detected by H&E staining, PCR, IF, EM immunohistochemistry, while virus isolation was used to detect virus particles. Acute and chronic vasculitis, signs of chronic endocarditis, with mononuclear cell accumulation and a fresh thrombus was found. Herpes viruses have already been thought to initiate cardio-vascular disorders, now this paper shows that a bovine gamma-herpes virus could also be a causative agent of vascular lesions in mammals fed a normal diet. BoHV-4-infection of rabbits could serve as a useful animal model for research into virus-induced human cardio-vascular diseases.     

Adherence to various host cell lines of Mycoplasma bovis strains differing in pathogenic and cultural features

Mycoplasma bovis is known to be responsible for pneumonia and arthritis in calves, as well as mastitis in dairy cows. Despite clear evidence of its pathogenic potential, little is known about mechanisms of cytadherence and the molecular factors involved. The purpose of this work was to compare adherence rates of M. bovis field strains to different host cell lines and study the effects of cloning and sub-culturing M. bovis strains on their adherence properties. Eighteen metabolically labeled M. bovis strains isolated from different pathological backgrounds were examined in adherence trials using four different host cell lines, i.e. embryonic bovine lung (EBL), embryonic bovine trachea (EBTr), Madin Darby bovine kidney (MDBK) and rabbit kidney (RK) cells. Although large interstrain variations in adherence rates (3.4-19.1%) were measured they could not be correlated to the pathological background (pneumonia, arthritis or mastitis). Adherence rates to the fibroblast cell line (EBTr) were significantly lower than those to the three epithelial cell lines (EBL, MDBK and RK). The only non-pathogenic strain (221/89) exhibited lower adherence rates than three isolates from clinical mastitis. Interestingly, adherence rates were significantly reduced after in vitro passaging. In contrast, no effect of single cloning of strains on adherence was observed. There was no general correlation between expression of variable surface proteins (Vsps) as monitored by immunoblotting and adherence rates, although alterations in Vsp expression profiles were seen as a consequence of passaging. As there is probably a large number of adhesins, variable and non-variable, on the surface of M. bovis cells the issue is very complex, and the most active components have yet to be identified.     

Effect of freezing goat milk samples on recovery of intramammary bacterial pathogens

With the aim of evaluating the effect of freezing goat milk samples on recovery of intramammary pathogens, 1200 milk samples from udder halves with subclinical intramammary infection were studied. Samples (20 ml) were frozen at -20 and at -80 degrees C. Thawing was carried out at room temperature at 7, 14, 21, 28, 58, 118, 178, 236 and 730 days after collection and bacteriological analyses were carried out to determine the number of colony forming units/ml (CFU/ml). Mixed model statistical analysis showed that bacterial group, temperature of storage, interaction of bacterial group and temperature of storage and the interaction of bacterial group, time and temperature of storage were statistically significant effects. For coagulase negative staphylococci (CNS), least squares means of log CFU/ml recovered at -20 and -80 degrees C were not different. Nevertheless, for Gram negative bacilli (GNB) a significant decrease was detected in samples frozen at -20 vs. -80 degrees C. At both temperatures and at different times of storage, significant increases were detected between log CFU/ml of CNS and values on day zero. At -20 degrees C, a significant decrease in GNB recovery was detected between freezing days zero and 730. This difference was not detected when goat milk samples infected by GNB were frozen at -80 degrees C. The results show that frozen milk samples can be useful in goat subclinical mastitis control programs.